Following this I surmise Professor Weiss read the Montagnier
interview. In the meantime, believing that Weiss would not reply
(mistakenly and I must give Professor Weiss full credit for being the
only HIV protagonist who has taken the time to debate us), I emailed Rex
Ranieri, a documentarian from TV Channel Nine in Australia. He emailed
Weiss on 29th March:
Dear Professor,
I have been observing the HIV/AIDS debate with some interest and I
have recently been contacted by Val Turner. He have sent me some
questions which he has put to you recently together with the subsequent
email discussion .
It appears to me that the argument for the existence of HIV is not
sufficiently rigorous.
Is there a bigger story developing here?
I look forward to your reply
Kind regards,
Rex
Rex Ranieri
Channel 9
Perth, Western
Australia
rranieri@perthtv9.net.au
+61 8 9449 9999
Fax +61 8
9449 9905
Mobile 0411 258344
Professor Weiss replied immediately:
If HIV does not exist, then neither did smallpox virus (variola), nor
does polio virus, tobacco mosaic virus in plants, etc. etc. If you wish
to deny the existence of viruses, and virus diseases, go ahead, but
leave scientists like me out of the picture.
Robin A Weiss
To which Rex Ranieri replied:
Professor,
My understanding is that Dr. Turner and his colleagues have
questioned not whether a number of other viruses exist. Only HIV. As far
as I am aware, a scientist does not prove that a particular virus exists
by pointing to the existence of others.
I am well versed with some of the argument so far (for a lay person)
so naturally your response contributes little to my questions.
I appreciate your desire to be left "out of picture" however as you
are a world renowned researcher who has spent some time on the question,
it is difficult for me to accept that you can bow out of the
discussion.
Naturally, It is your perogative not to respond, however I think that
this would ultimately be damaging to both sides of the argument. We have
seen many examples of media debacle which can result from lack of
discussion.
I realise that your time is valuable and I urge you to respond to my
questions. Please accept that my intentions are to arrive at the truth,
whatever that may turn out to be.
Kind regards,
Rex
Weiss responded to this email approximately two months later (see
Addendum III).
Meantime I sent reminder emails to Weiss. Eventually he replied:
March 26th
I am very tied up with work at present and will give you a considered
reply in due course.
Robin A. Weiss
I responded:
Dear Professor Weiss,
Thank you for your reply. I fully understand that you have been busy
moving from Fulham Road to Clevland Street and setting up your new
department. I am also sure that you understand how anxious both my
clinical and non clinical colleagues and I are to examine your
considered reply.
Yours sincerely,
VF Turner
After more reminders Weiss emailed me:
April 15th
Dear Dr Turner
You have breached my correspondence with you as an academic colleague
by forwarding it to a journalist, Rex Ranieri. In your message to him,
you write that that you do not hold any great hope that I shall answer a
second time, and yet to me you express your understanding that I'm busy
with other things. So here is my second response. I hope it is my last
response, because I find the issue of 'purification' quite sterile, and
unconnected with matters of medical importance. We are simply talking at
cross-purposes.
There appears to be a consortium of medical people and biophysicists
in Perth who have a fixation of HIV purification. Perhaps you are
influenced by this. It is also a 'cause' championed by the British based
magazine 'Continuum' founded by Jodi Wells whom I knew and who sadly
died of an AIDS-like disease a few years ago. He initially supported
Peter Duesberg's view that HIV indeed exists and can be purified, but
that it is harmless. Then he shifted into a denial that there is any
such thing as HIV. With Harold Jaffe I argued against Duesberg's view in
Nature nearly 9 years ago (Nature 345: 659-660, 1990). I've nothing more
to say on this issue, save that with the efficacy of combination
anti-retroviral drugs, Duesberg seems to have lost his constituency of
support among 'lay' gay men.
Now, to turn to your points sent on March 10 regarding my first
reply:-
1. and 2. You confuse isolation and purification. I see no
contradiction between what I wrote - in 1986 or in 1999 - and what
Barre-Sinoussi and colleagues had reported previously. One can isolate
some viruses by propagating them in cells in culture. For example, HIV,
smallpox virus, measles virus, polio virus. There are other viruses
which no-one has yet succeeded in serially propagating in culture
following isolation because they require specialized, differentiated
cells; for example, hepatitis B virus, human papilloma virus types 16
and 18 (associated with cervical cancer), and so on.
In both cases, viral genomes can be isolated, indeed highly
'purified' by molecular cloning using recombinant DNA methodology. Thus
it is almost routine now in our lab and may other research labs to clone
the HIV genome as DNA in bacterial vectors, and then recover them again
in infectious form by transferring that DNA back into human lymphocytes.
It is difficult to conceive anything 'purer' than the complete cloned
virus without any proteins, particles, etc.
3. I do not think Barre-Sinoussi et al misled the scientifically
community by calling the 1.16 g/ml band purified virus. But if you and
your colleagues prefer to call it enriched but not yet completely pure,
I would happily concur with that opinion. This illustrates what I mean
by a sterile argument: how pure is pure? Is distilled water 'pure'? Yes,
but it will still have a few parts per million or per billion of other
soluble molecules.
Are your surgical instruments sterile? Yes regarding bacteria and
viruses, if they have been heat-treated or autoclaved. No, regarding the
agent of Creuzfeld-Jakob disease which partially resists such treatment.
Yet, every surgeon knows what others mean by sterile. Let's not get
bogged down in how pure is pure.
The important thing is serial propagation of the microbe. Koch and
Petri over 700 [70] years ago 'purified' bacteria by propagating them as
colonies (clones) on gelatin in Petri's dishes - nowadays we use
agar-agar with nutrients in place of gelatin. Did Koch purify the
microbes. Yes in his and my terms, maybe not in yours. Certainly he did
purify [?not] them by biophysical methods such as sucrose gradients, but
nothing else kept reproducing itself on the 'impure' nutrients. So it is
the same for viruses. As intercellular [intracellular] parasites, of
course, they can only be propagated in living cultured cells (or in
plants, animals or humans) but one can 'plague-purify' them - a term
dating from early bacteriophage studies in the 1920s. Animal viruses
were similarly plague-purified: polio in 1952; vaccinia around 1955. We
used a plaque 'purification' or biological cloning technique for HIV in
1989. No, these were not physically pure, but they were biologically
pure, ie they were cloned. Molecular cloning, however, as I mentioned
already is one step better. Both methods to my mind, are sufficiently
purified to draw scientific conclusions, although one must be cautious
not to draw conclusions beyond the validity of the data, including the
kind of purity, biological, molecular, chemical or physical.
4. My definition of isolation of HIV by Barre-Sinoussi et al. Gallo
and Levy and others in the early days of AIDS research is propagation in
culture. Today, however, we more often use molecular cloning, then
recover the cloned genome or partial genome and characterise its
phenotype.
5. Yes, I do claim that visualization of HIV by electron microscopy
was, in 1983/84, an important component of the collective data on virus
isolation. Taken together with virus propagation, reverse transcriptase
activity and enrichment of particles by isopycnio [density] gradients,
it convinced me that HIV is a retrovirus. Even more so, it was
Montagnier's electron micrographs published in April 1984 and previously
shown at Cold Spring Harbor Laboratory in September 1983 that convinced
me that HIV was probably a lentivrius among retroviruses, as they
resembled particles of equine infections anaemia virus - a lentivirus
first propagated by inoculating a filtrate too small for bacteria to
pass through into horses and donkeys and causing disease. So yes, I am
definitely arguing that, disregarding your meaning of 'purification',
but with my meaning of 'isolation', you can make quite large amounts of
HIV proteins and smaller amounts of RNA.
6. Barre-Sinoussi et al published electron micrographs of early
budding forms of virus only, that were not immediately identifiable as
retrovirus particles. By September 1983, Montagnier's electron
micrographs looked more typical.
7. There are many different tests for HIV-specific antibodies.
Today's commercial test kits are based on oligopeptides and on proteins
manufactured from cloned HIV DNA. No biological test for anything is
100% specific and 100% sensitive, but today's HIV tests are as good as
tests for any other human viral pathogen. Likewise, today's PCR primers
are highly specific and sensitive for the major strains of HIV in
developed countries. Some 'outlier' strains, especially in Gabon and
Cameroon are not picked up quite as sensitively and therefore estimates
of viral load with these 'outlier' infections should be interpreted
cautiously.
There will always be a few people who cannot be convinced by the data
before our eyes - or who emotionally wish to deny what the rest of us
regard as facts. Of course, interpretation will change over time.
Newetonian physios still serves pretty well for the dynamics of road
accidents, but Einstein's relativity superseded it on a cosmic scale. In
my view, to deny the existence of HIV is a bit like denying the Nazi
holocaust.
If we are to doubt HIV as a cause of AIDS, we must cast even more
doubt on variola as a cause of smallpox, and the existence of measles,
mumps, influenza and respiratory syncytial virus. None of these would
pass your definition of purification. None of these has been 'purified'
even by culture propagation (my sense) to the extent that has been
achieved for poliovirus and for HIV.
This terminates our debate.
Robin A Weiss
Although Professor Weiss terminated the debate, my colleagues and I
were far from satisfied. Eleni Papadopulos and I spent many weeks
preparing a response (see below). This was sent in late July with the
following note:
22/7/99
Dear Professor Weiss,
I apologise for the length of the attached file [35 pages] but it is
impossible to debate this topic without data and citations. I trust you
appreciate this necessity.
I greatly appreciate your reply to the questions in my last email.
Let me say that eighteen years ago neither I nor my colleagues in the
Perth Group set out to frustrate the efforts of scientists such as
yourself. Rather, we began as you and many others did in the early
1980s, to make a contribution to solving the problem of AIDS. Certainly
we are poles apart from the mainstream but, as Philebus says in the
Dialogues of Plato, I hope "we are not simply contending in order that
my view or that of yours may prevail, but I presume we ought both of us
to be fighting for the truth"..
I am disappointed that you do not intend to continue the exchange but
I respect your decision. Sometime in the near future I intend to put our
debate on the Perth group website. Thus, if you would like to add or
alter your replies in any way I will encompass these in the posting. If
you would like to contribute anything else at all, scientific or perhaps
philosophical including your views on dissent and dissenters, I would be
very pleased to post these as well.
I look forward to hearing from you once again.
Yours sincerely,
VF Turner
31/8/99
Dear Dr Turner
I have been in your country during August without opening my e-mails.
We shall have to agree to differ on the nature and existence of HIV. I
have no additional comments to make for your website.
Robin A Weiss
My response to this was:
1/9/99
Dear Professor Weiss,
Thank you for your email. I am truly sorry you are unwilling to
continue the debate. Eleni Papapdopulos and I spent several weeks
preparing what I consider a worthy reponse and believe it your
responsibility to answer the points I have raised. This especially
applies to the arguments related to the extant HL23V. As far as I can
tell evidence for the the "isolation" and thus the existence of HL23V is
better than that for HIV. Yet HL23V has disappeared from the scientific
literature and no longer exists.
I am disappointed you did not look us up while you were in Australia.
My colleagues and I would have been delighted to take you out for a meal
in Perth's Kings Park overlooking the Swan River. We have some excellent
wines which I am sure you would have appreciated. We would have enjoyed
your company and the invitation is open should you ever return.
I would like to thank you once again for your time and effort on
behalf of this debate. This is not a polemic about any particular person
winning. Any fighting is about "fighting for the truth".
Yours sincerely,
Val Turner
REPLY TO PROFESSOR R WEISS BY DR. VF TURNER OF THE PERTH GROUP
JULY 21ST 1999
Dear Professor Weiss,
Scientists should be particularly hard on hypotheses. An hypothesis
must explain and predict everything. For example, the HIV theory has to
explain why, in Western countries, HIV/AIDS has remained in the original
risk groups. It is claimed that HIV/AIDS is spread predominantly by
sexual contact of one kind or another and "everyone is at risk" from "a
virus that does not discriminate". But prostitutes who do not use drugs
are not infected with HIV. In Australia for example, a decade after the
beginning of the AIDS era, the number of prostitutes infected by sexual
intercourse, despite their being "seriously at risk of HIV infection",
was zero. In 1994, Manaloto reported on the "Natural history of HIV
infection in Filipino females commercial sex workers". Over seven years
72/53,903 (0.01%) were found to be HIV positive and "Intravenous drug
use was denied in all cases". We can take a step further and examine an
instance where we can be sure of unprotected sexual intercourse in
heterosexuals, that is, new born babies. Again in Australia, in 1989 a
study testing 10, 217 blood samples of newborn babies found no babies
and thus presumably none of their mothers or fathers HIV positive. These
data are supported by Nancy Padian and colleagues’ ten year study of
heterosexual couples (1986-1996). There were two parts to this study,
one cross-sectional, the other prospective. Of the HIV negative male
partners of 82 positive female cases only 2 were found HIV positive but
under circumstances considered ambiguous by Padian. In the prospective
study, starting in 1990, 175 HIV-discordant couples were followed for
approximately 282 couple-years. At entry, one third used condoms
consistently and in the six months prior their last follow up visit, 26%
of couples consistently failed to use condoms. There were no
seroconversions after entry including the 47 couples not using condoms
consistently. Based on the 2/86 men who became HIV positive in the early
study, the risk to a non-infected male from his HIV positive female
partner was reported to be in the order of 1/9000 per contact. From this
statistic one can calculate that on average, a male would need to have
6000 sexual contacts with an infected female to achieve a 50% chance of
becoming HIV positive. If sexual intercourse were to commence at age 20
and average three times weekly, this would occupy a lifetime; If
heterosexual men so rarely become infected how do heterosexual women
become infected?
Despite the fact that HIV/AIDS experts continuously assert that
epidemiology proves HIV causes AIDS, in my view epidemiological data are
incapable of proving anything . But such data may prove scientifically
useful when inconsistent with the predictions of a particular theory. As
the above data amply illustrate. Thus, in my view and our Group, unless
we entertain extraordinary properties for a putative infectious agent,
it is obligatory for scientists to question the HIV theory of AIDS. Or,
as Simon Wain-Hobson expeditiously put it, "a virus's job" is to spread.
"If you don't spread, you're dead". I could not agree more. Inevitably,
an exercise of this sort leads Homo sapiens to question what the
antibody and genomic tests are measuring. How have their specificities
for a unique, infectious agent been determined and what are they? Since
these tests purport to use "HIV" proteins and RNA (cDNA) as diagnostic
reagents, and since viruses are obligatorily cultured in cells where one
would expect to find hundreds if not thousands of cellular proteins,
RNAs and DNAs, we would expect there to be proof that HIV had been
purified. Otherwise it would be impossible to claim that certain
proteins, RNA or DNA are unique constituents of a retrovirus HIV.
However, the data of Montagnier, Bess and Gluschankof, all HIV
protagonist experts, prove there is no such thing as purified HIV and,
according to you, "purification is not particularly important" and
Montagnier was right "to say it was not purified virus". These being the
case we are left wondering on what basis scientists such as yourself
accept that there is such a retrovirus as HIV. This, of course, is what
you and I are presently debating. I am sure we can at least agree that
without HIV there can be no HIV theory.
Since your definition and understanding especially, but not only of
purification, isolation, virus, viral cloning are different from mine, I
concur we are "talking at cross-purposes". Where we do seem to agree is
on the importance of medical issues, although we seem to disagree
exactly what constitutes these medical issues. You state, "I find the
issue of purification quite sterile, and unconnected with matters of
medical purposes". However:
(a) for more than 15 years
(i) the world has been made to believe that Barré-Sinoussi,
Montagnier and their colleagues claim to have proven the existence of a
unique retrovirus, HIV, by purifying it using "isopycnic gradients";
(ii) in 1997 Montagnier admitted that the 1.16g/ml band in their
isopycnic gradients which they claimed to have proven was "purified
labelled virus" did not contain even retrovirus-like particles. Yet,
this "purified" virus is accepted as being the cause of the black plague
of the century;
(iii) in the name of this virus hundreds of thousands of individuals
have died and the lives of millions of people have been adversely
affected either directly or indirectly.
Surely these are "matters of medical importance"?
(b) According to Montagnier, "analysis of the proteins of the virus
demands mass production and purification. It is necessary to do that.
And there I should say that we partially failed". In fact I venture that
Montagnier totally failed since the proteins and the RNA which he and
his colleagues named HIV proteins and RNA were taken from an isopycnic
gradient (the 1.16g/ml band) which they called "purified labelled
virus", but where, even after a "Roman effort", they could not find even
particles with the "morphology typical of retroviruses", much less
retrovirus particles, not to mention a specific retrovirus, HIV. The
claim that some of the proteins in this "purified labelled virus" were
HIV proteins defies not only scientific reasoning but commonsense. This
is no different from a fisherman claiming to have processed a catch of a
totally new species of fish into unique fish proteins when his net
contained anything but "fish-like" objects, or me as a surgeon going
into an operating room where there are many types of animals, but none
which look human, removing a few hearts and kidneys, transplanting them
into humans in an adjacent operating room and claiming the transplanted
organs were of human origin. Yet:
(i) In the name of one of these "HIV proteins", reverse
transcriptase, (based solely on the detection of which many researchers
publish papers claiming "HIV isolation"), hundreds of thousands of
individuals have been treated for more than 10 years with toxic
drugs;
(ii) In the name of another of these "HIV proteins", protease, in the
last few years even more toxic "cocktails" of drugs have been used;
(iii) Since 1984, in the name of yet another "HIV protein", gp120,
hundreds of millions of dollars have been spent to develop a vaccine
which it is said will be especially useful in the developing countries
where the majority of AIDS cases are tuberculosis. It is the first time
in medical history that a vaccine based on a protein said to be a
retroviral protein, but which never has been shown to be a constituent
of even a retrovirus-like particle, is going to be used to prevent a
disease caused by a bacterium.
(iv) In the name of an antibody test employing these "HIV" proteins
as antigens, millions of people have been told that they are infected
with a lethal retrovirus;
(v) A stretch of RNA (or its cDNA) which never has been shown to be a
constituent even of a retrovirus-like particle much less of a retrovirus
is used as a hybridization probe and PCR primer to prove infection with
a retrovirus claimed to be deadly, and in fact to quantify it.
Surely these also constitute "matters of medical importance"?
For physicians like me, stranded between laboratory scientists and
patients, the meaning of the antibody test and the "viral load" are of
extreme "medical importance". And whether they are aware or not, for
"lay" gay men, drug users, haemophiliacs, blood transfusion recipients,
pregnant women and their offspring, and all their relatives, these are
also matters of extreme "medical importance".
As far as I am aware there are two groups of "anti-retroviral drugs".
The reverse transcriptase inhibitors which are said to prevent the
reverse transcription of "HIV RNA" into "HIV DNA", and the protease
inhibitors which are said to render particles non-infectious. In other
words, the effect of both groups of drugs is to decrease the synthesis
of new "HIV DNA", that is, to decrease the amount of provirus. Since
none of the "anti-retroviral drugs" used to date has any effect on the
transcription of "HIV DNA" into "HIV RNA", a decrease in the latter, the
"viral load", must always be preceded by a decrease in the former (the
"viral burden"). This is not the case. While many studies report
"anti-retroviral" treatment decreases the viral load from millions to
undetectable levels, the "HIV DNA" remains constant. In a study
published this year "of 34 patients on combination therapy who had
plasma HIV-1 RNA levels of less than 200 copies/ml,
replication-competent virus was isolated" from 32 (94%). In longitudinal
studies it was shown that the frequency of isolation did not depend on
the length of combination therapy or the time it was started. Compare
this with the 37% frequency of isolation by the best HIV laboratories of
non-treated patients in a 1994 WHO study. This can be explained by one
or more of the following:
(a) The proviral theory is wrong, that is, there is no relationship
between "HIV DNA" and "HIV RNA"
(b) Either "HIV DNA" or "HIV RNA" or both have nothing to do with a
retrovirus;
(c) The drugs have no anti-retroviral effect, they only mask the
measurement of "HIV RNA".
I venture to say that your present enthusiasm for "combination
anti-retroviral drugs" is matched only by your former enthusiasm for
AZT. However, there are at least two reasons why this drug should never
have been introduced into clinical practice and while its continued use
should be abandoned. First and foremost, the pharmacology of AZT proves
beyond all reasonable doubt that AZT cannot act as a reverse
transcriptase inhibitor, DNA chain terminator. Secondly, even before the
AIDS era it was known that AZT is very toxic.
The Concorde study, the only truly clinical blind study, has
vindicated both John Lauritsen's and Peter Duesberg's claims that AZT
instead of saving lives may endanger them.
In another study published this year by Italian researchers, the
authors followed up, for the first three years of life, HIV seropositive
children born to mothers who either did or did not receive AZT
treatment. The two groups of children were similar with respect to all
variables (year of birth, maternal clinical condition, birth weight and
treatments) apart from age at the beginning of PCP chemopropylaxis,
which was undertaken earlier in these children who were born to mothers
who took AZT. They found that the children born to these mothers "had a
higher probability of developing severe disease" (57.3% versus 37.2%) or
severe immune suppression (53.9% versus 37.5%) and a lower survival
(72.2% versus 81%)".
Given these findings any disinterested scientist, clinician, layman,
even politician would advise against the administration of AZT to
pregnant women. But not HIV/AIDS experts. The authors of this study
concluded: "Findings may suggest a need to hasten HIV-1 diagnosis in
infants of ZDV-treated mothers and undertake an aggressive
anti-retroviral therapy in those found to be infected", and "should not
be misinterpreted as a reason not to use ZDV prophylaxis, which is
effective in preventing perinatal HIV-1 infection". However, since as
the authors of this study claim, "A strong association exists between
high maternal viral load and an increased risk of transmission" and
since none of the many studies conducted to date has shown a decrease in
viral load by AZT, it follows that AZT cannot inhibit maternal
transmission. In the face of such data the authors' conclusions and
recommendations amply illustrate to what lengths some scientists will
strive not to question the HIV theory. If you can offer any other
rational explanation I would be most grateful to hear it.
According to Professor Brian Gazzard, President of the British HIV
Association, treatment with AZT monotherapy is "ludicrous". Yet not only
is AZT monotherapy prescribed to mothers and mothers who object to this
treatment for their children are threatened with having their children
taken from them. Your enthusiasms for combination therapy is shared
neither by the physicians who prescribe it nor by the "lay" gay men who
take it. "As physicians venture into even wilder frontiers of HIV
treatment, the grand experiment with combination therapies, called
Highly Active Anti-Retroviral Therapy, or HAART, is rushing forward
without any data. No-one is keeping track". Apparently one of the few
physicians, if not the only one who tries to make any sense of HAART is
Michael Saag who supervises research and the care of more than one
thousand AIDS patients in Birmingham, Alabama. "In one year, 157 of
Saag's patients collectively took 189 different drug formulas, with only
three patients taking the same mix of HAART drugs…despite such rigorous,
individualized medical attention, Saag says, the HAART 'dam' is already
leaking and there's high danger of it collapsing altogether...Failures
are occurring right and left".
According to Dr Wafaa El-Sadr from Harlem Hospital "it takes a heap
of denial to reach anything but sobering, even grim conclusions"
regarding HAART. Let me also quote a few gay men, who, unlike the
courageous Jodi Wells and the equally courageous and well informed
present editors of Continuum, are not in "denial" regarding the
existence of HIV. "Anything they give me, I suspect would simply create
more side effects and then fail after only a few months, leaving me in
even worse shape than I'm now. As my doctors see it, being of therapy is
doing nothing, and doing nothing is what scares them most...So when they
tell you all is well, they may be doing it less for you than for
themselves" (Stephen Gendin POZ Magazine, January 1999). "None of us
would say that having access to the best available scientific
information every step of the way has helped us
particularly...professional judgement has consistently been way out in
front of evidence. We're going to have to rethink everybody". Saag
agrees: "We don't know what we are doing. Hubris! Hubris! We need to be
scientists every step of the way and do our darndest to seek what
reality is". If the claims for actual reality of HIV are also based on
professional judgement rather than evidence we are indeed guilty of
arrogant presumption and must expect nemesis.
Thus, in your position, I would not uphold "anti-retroviral drugs"
either in support of the HIV theory of AIDS or the existence of HIV. The
only rational conclusion one can draw from the "anti-retroviral"
treatment data is either there are no such drugs or there is no such
thing as a retrovirus, HIV. (Additional data on PIs is at the end of the
reference section*)
Bearing all this in mind, I would like to comment point by point on
your answers regarding claims for the existence of HIV.
ANSWER 1 AND 2
My questions were:
1 Why, in 1986, did you and your colleagues write: "A so-called AIDS
virus isolate was first reported in 1983 by Montagnier and his
colleagues in France who named the material "Lymphadenopathy Associated
Virus One". Did or did not Montagnier isolate, purify a retrovirus?
2. If he did not why did you say that he had? If you were aware that
he did not do this, and this was the reason for you using the word
"material" to describe his finding, why did you not, as a well known and
respected retrovirologist, draw the attention of the rest of the
scientific community to it, especially if one considers the extremely
important consequences?
Your answer was:
You confuse isolation and purification. I see no contradiction
between what I wrote - in 1986 or in 1999 - and what Barre-Sinoussi and
colleagues had reported previously. One can isolate some viruses by
propagating them in cells in culture. For example, HIV, smallpox virus,
measles virus, polio virus. There are other viruses which no-one has yet
succeeded in serially propagating in culture following isolation because
they require specialized, differentiated cells; for example, hepatitis B
virus, human papilloma virus types 16 and 18 (associated with cervical
cancer), and so on. In both cases, viral genomes can be isolated, indeed
highly 'purified' by molecular cloning using recombinant DNA
methodology. Thus it is almost routine now in our lab and many other
research labs to clone the HIV genome as DNA in bacterial vectors, and
then recover them again in infectious form by transferring that DNA back
into human lymphocytes. It is difficult to conceive anything 'purer'
than the complete cloned virus without any proteins, particles, etc.
My response:
I do not accept isolation and purification as being different. The
word "isolation" means to place apart or alone. (From Latin "insulatus",
made into an island). "Purification" means to obtain something free from
impurities. For a particle such as a virus, isolation and purification
are the same thing. The act of placing an object apart or alone,
isolation, is totally different from multiplying, transmitting, that is
propagating that object. The fact that something can be multiplied or
propagated through many generations of cultures neither means that it is
any more or less isolated nor that you are any more aware of its
identity. No object including a virus can be isolated by propagation.
You, more than anybody else, know that there are some very significant
differences between retroviruses and other viruses. The subject of
interest for you and me is HIV. It is of no interest what methods have
been used to prove the existence of other viruses and whether they are
good or bad. What is sufficient for one group of viruses may not be
sufficient for another. What we all want to know is whether the methods
and the evidence which are claimed to prove the existence of HIV and
thus of its proteins and its genome, do so. To claim propagation of a
unique retrovirus, HIV, one must first prove its existence which can be
done only by isolating, purifying it. To date, nobody, not even you, has
achieved this.
By molecular cloning it is meant the production of identical copies
of, for example, a DNA molecule, any DNA molecule, from an ancestral
molecule by splicing it into a suitable cloning vehicle, for example a
bacteriophage. The DNA molecules can be either of viral or cellular
origin. Or they could be artificial. The molecules within the
bacteriophages can, in their turn, be introduced in any cells. If you
want to call these molecules an "infectious form" (I will not) then you
must accept that any piece of DNA, no matter what its origin, is an
"infectious form".
Unquestionably, by molecular cloning one can obtain large amounts of
"purified" DNA, but equally unquestionable is the fact that molecular
cloning tells you nothing about the origin of a particular DNA molecule.
To clone "the HIV genome" one must know beforehand that a particular DNA
is the HIV genome. In other words, one must start with "HIV RNA"
which can be obtained only by prior purification of particles proven to
be retroviral particles. Since this has not been achieved, then what is
the origin of the DNA fragment which is known as the "HIV genome" which
you routinely clone in your lab? If you are using "material" similar to
that used by Montagnier in 1983, in which there is no proof there were
even retrovirus-like particles, then surely nobody, not even you, can
claim this RNA (cDNA) is the genome of a retrovirus, not to mention a
unique retrovirus, HIV.
As an undergraduate medical student I was taught that viruses are
infectious particles of particular morphologies containing nucleic
acids, proteins and lipids. According to all other retrovirologists
including Montagnier, Gallo and Gelderblom, retroviruses are particles
with a diameter of 100-120nM containing "condensed inner bodies (cores)"
and surfaces "studded with projections (spikes, knobs)". If, unlike
other viruses, HIV consists of nothing else but an "infectious form" of
DNA molecules (as you claim: "It is difficult to conceive anything
‘purer’ than the complete cloned virus without any proteins, particles,
etc"), then why do all other retrovirologists, without exception, claim
that the gp120 protein is absolutely necessary for HIV infection? If HIV
is nothing else but a pure nucleic acid molecule (RNA) "without any
proteins", then why is there so much effort, not to mention expense, to
develop a vaccine based on the "HIV" proteins?
ANSWER 3
My question was:
In 1983, when B-S et al published their paper entitled, "Isolation of
a T-lymphotrophic retrovirus from a patient at risk for acquired immune
deficiency syndrome (AIDS)", and called the 1.16g/ml band "pure labelled
virus", did they mislead the scientific community?
Your answer was:
I do not think Barre-Sinoussi et al misled the scientific community
by calling the 1.16 g/ml band purified virus. But if you and your
colleagues prefer to call it enriched but not yet completely pure, I
would happily concur with that opinion. This illustrates what I mean by
a sterile argument: how pure is pure? Is distilled water 'pure'? Yes,
but it will still have a few parts per million or per billion of other
soluble molecules. Are your surgical instruments sterile? Yes regarding
bacteria and viruses, if they have been heat-treated or autoclaved. No,
regarding the agent of Creuzfeld-Jakob disease which partially resists
such treatment. Yet, every surgeon knows what others mean by sterile.
Let's not get bogged down in how pure is pure. The important thing is
serial propagation of the microbe. Koch and Petri over 70 years ago
'purified' bacteria by propagating them as colonies (clones) on gelatin
in Petri's dishes - nowadays we use agar-agar with nutrients in place of
gelatin. Did Koch purify the microbes. Yes in his and my terms, maybe
not in yours. Certainly he did purify them by biophysical methods such
as sucrose gradients, but nothing else kept reproducing itself on the
'impure' nutrients. So it is the same for viruses. As intracellular
parasites, of course, they can only be propagated in living cultured
cells (or in plants, animals or humans) but one can 'plaque-purify' them
- a term dating from early bacteriophage studies in the 1920s. Animal
viruses were similarly plaque-purified: polio in 1952; vaccinia around
1955. We used a plaque 'purification' or biological cloning technique
for HIV in 1989. No, these were not physically pure, but they were
biologically pure, ie they were cloned. Molecular cloning, however, as I
mentioned already is one step better. Both methods to my mind, are
sufficiently purified to draw scientific conclusions, although one must
be cautious not to draw conclusions beyond the validity of the data,
including the kind of purity, biological, molecular, chemical or
physical.
My response:
We cannot call Barré-Sinoussi's 1.16g/ml band pure or even "enriched"
since the scientists who prepared the material conceded it contained no
retrovirus-like particles, much less the particles of a unique
retrovirus. Perhaps it is inconceivable to you that Montagnier made this
admission. In that case let me present you with the actual transcript
from the videotape given to me by Djamel Tahi, the French investigative
journalist who conducted the interview at the Pasteur Institute in July
1997. Montagnier was asked, "Pourquois les photographies du EM publiées
par vous, proviennent de la culture et non de la purifcation?"
Montagnier's reply was: "Il y avait tellement peu de production que
c’était impossible de voir soit dans un culot de virus à partir d'un
gradient. Il n'y avait pas assez de virus pour faire ça. Bien sûr on l'a
cherché, on l'a cherché aussi dans les tissus de départ, de la biopsie
également. On a vu des particules, mais elles n'avaient pas la
morphologie typique des retrovirus. C’était trés différent...
relativement différent. Donc avec la culture il a fallu beaucoup
d'heures pour trouver les première images. C’tait un travail de
romain".
Consider your analogy of pure water. Suppose you are doing a vital
experiment which requires pure water. Your colleagues on the floor above
provide a bottle labelled "pure water" which forms a critical part of
your experiment and ultimately the results of this experiment are used
in a way which affects the lives of millions of people. Fifteen years
later you are informed that the bottle your colleagues gave you, far
from being pure water, contained not even one molecule of water. Would
you not think that your colleagues were wrong to label it "pure water"
and that they had misled you? Would you then be happy to call it water
enriched?
It is not valid to argue that the technique of Koch and Petri
justifies the method used to prove the existence of HIV. They did not
define purification or, if you like, isolation as "propagation in
culture", as you do. They did not "purify them" by biophysical methods
such as sucrose gradients because this method is a much later invention
and is not even necessary. In their cultures Koch and Petri had gelatin
plus particles which could be unambiguously identified as bacteria using
the light microscope. Any proteins (other than gelatin) RNA or DNA found
in the culture could only be of bacterial origin. Any biological effect,
including a plaque produced by an aliquot from this culture which is not
produced by gelatin alone must be induced by the bacteria. But it is not
the "same for virus", especially retroviruses. Unlike the bacteria,
dishes which in addition to bacteria contain only one known protein,
gelatin, the "HIV" dishes contain cells, cellular fragments and perhaps
bacterial fragments as well as mycoplasmas and viruses. As far as the
presence of HIV is concerned, it is sufficient to mention that some
electron micrographs of culture supernatants have shown some particles
with some of the morphological characteristics, but no particles with
all of the morphological characteristics of mature retroviruses.
Even assuming that the particles were those of a unique retrovirus,
the proteins and RNA present in the culture or even the culture
supernatant cannot be assumed to belong to the virus. Induction of a
biological effect by the culture supernatant including "plaque" need not
be due to the virus, but to any of the many other factors present. I am
not aware of any publications describing "plaque 'purification' or
biological cloning technique of HIV in 1989". I would be interested to
know how one can know that a "plaque" or any other biological effect is
induced by a specific object without prior proof for its existence.
"Plaque" and other biological effects are induced by addition and
interaction while isolation (purification) is a process of subtraction
and separation. Biological effects including plaque can be used only to
detect a retrovirus, and then, if and only if, there is a prior
knowledge they are specific to the retrovirus, which can be done only by
first isolating the virus.
I agree with you that one must be cautious not to draw conclusions
"beyond the validity of the data". However, unlike you, I believe that
the claim for the existence of HIV is based on interpretations which go
far "beyond the validity of the data".
ANSWER 4
My question was:
Since it is generally accepted, and makes common sense, that the
existence of a new retrovirus can be proven only by isolating it (both
B-S and Gallo claimed to have proven the existence of HIV by isolating
it) what is the scientific basis for your claims that B-S et al
discovered a new retrovirus?
Your answer was:
My definition of isolation of HIV by Barre-Sinoussi et al. Gallo and
Levy and others in the early days of AIDS research is propagation in
culture. Today, however, we more often use molecular cloning, then
recover the cloned genome or partial genome and characterise its
phenotype.
My response:
In 1983 Barré-Sinoussi et al, in the T cell culture derived from the
lymph nodes from a patient at risk of AIDS, detected reverse
transcriptase activity. The cells from this culture were cultured with T
cells from a healthy donor whereupon the detection of reverse
transcriptase activity in this second culture was considered proof for
isolation of a retrovirus which originated from the patient. Detection
of reverse transcription, even if detected in thousands of consecutive
cultures is not proof for the isolation of the enzyme reverse
transcriptase, much less the isolation of a unique retrovirus, HIV. The
measurement of cardiac or liver enzymes in cases of myocardial
infarction or hepatitis respectively cannot be construed as "isolation"
of the heart or liver. In fact, detection of reverse transcription is
not even proof of reverse transcriptase as other enzymes ably perform
the same task.
Reverse transcription is non-specific. Indeed, in 1996 you wrote:
"Now we know that a broader group of genetic elements than retrovirus
utilise reverse transcription at some stage of replication; these
include hepadnoviruses (including hepatitis B virus), cauliflower mosaic
virus and retrotransposons of eukaryotes and prokaryotes". If this is
the case, given the right conditions, one would expect to detect reverse
transcription in all cultures, irrespective of the presence or absence
of a retrovirus. Then, surely you would agree that the finding of
reverse transcription is not even proof for the detection of a
retrovirus, much less its isolation, even if one uses your definition.
In 1984, Levy et al cultured cells from patients with Kaposi's sarcoma a
disease which everybody agrees is not caused by HIV. The supernatants
were tested for RT activity, the cells for reactions with serum from
Barré-Sinoussi's patient BRU. Some cultures were examined by electron
microscopy. The finding of a positive result with "any of those tests"
was considered proof for HIV isolation.
In 1984 Gallo said that Montagnier's group data did not constitute
proof for isolation, and defined isolation as the detection of more than
one of the following:
"reverse transcriptase activity in supernatant fluids;virus observed
by electron microscopy [retroviral particles in the cultures];
intracellular expression of virus-related antigens detected with
antibodies from seropositive donors or with rabbit antiserum to
HTLV-III; or transmission of particles". By transmission of particles
was meant detection of reverse transcriptase or particles in cultures of
"human cord blood, bone marrow, or peripheral blood T lymphocytes",
cultured with concentrated fluids from the cell cultures from tissues
obtained from AIDS patients.
However:
1 As can be seen for Gallo et al, like for Barré-Sinoussi et al, the
detection of reverse transcription in two consecutive cultures is proof
for isolation, although a decade before the AIDS era Gallo himself
reported such activity in normal cell cultures. In the AIDS era such
activity was reported in the non "HIV" infected H9 cell line from which
Gallo "isolated" HIV as well as many other cell lines used to "isolate
HIV" and normal cell cultures.
2. Alternatively, according to Gallo's criteria, HIV can be
"isolated" in the absence of reverse transcriptase activity, considered
by Gallo to be the "sine qua non" of a retrovirus.
3. It is impossible for "rabbit antiserum to HTLV-III [HIV]" to
predate proof for the existence of, and pure HIV. It makes no sense at
all for Gallo to claim use of an antiserum to prove the existence of a
virus when the production of that antiserum is totally dependent on use
of the same, purified virus.
4. From the interaction of a protein with an antibody it is
impossible to determine the origin of the protein or the antibody. From
such a reaction one can determine the origin of only one of the
reactants, and then if, and only if, the origin of one of the reactants
is known and if there is proof that the interaction is specific. The
specificity of such reaction between the "HIV" proteins and antibodies
present in patient sera can be proven only by using HIV isolation,
purification as a gold standard. Gallo et al claim that the interaction
between proteins present in the cells and antibodies in the patient sera
proves not only that the proteins and antibodies are HIV protein and
antibodies, but the detection of such reaction together, for example
with reverse transcription proves HIV isolation. In fact, Gallo has
conceded that for him, an antibody test is the quintessence of "HIV
isolation". Interviewed in camera by Hugh Christie at the 1998 Geneva
AIDS conference he answered, "Sometimes we had Western blot positive but
we couldn't isolate the virus. So we got worried and felt we were
getting false positives sometimes so we added the Western blot [sic].
That's all I can tell you. It was an experimental tool when we added it
and for us it worked well, 'cos we could isolate the virus when we did
it". In other words, Gallo knew full well that to prove the specificity
of the antibody test it was essential to isolate the virus. When he saw
no correlation between antibodies and virus "isolation" Gallo resolved
the dilemma by defining virus "isolation" to include the Western blot
antibody test.
5. Both groups, Gallo’s and Montagnier’s, found proteins in
non-infected cells which reacted with sera from the AIDS, patients.
6. Gallo’s claim for proof of isolation that the findings of
particles in the culture supernatants of two consecutive cultures, or in
one culture together with reverse transcription or antigen/antibody
reactions is puzzling. A decade earlier, Gallo warned that "Release of
virus-like particles morphologically and biochemically resembling type-C
virus but apparently lacking the ability to replicate have been
frequently observed from leukaemic tissue", that is, these properties
are not proof for the existence of such viruses.
7. In the 1970s, such particles were frequently observed in human
leukemic tissues, cultures of embryonic tissues, and in the majority, if
not all, of human placentas. In the AIDS era retrovirus-like particles
have been revealed in non "HIV" infected H9 (the leukemic cell line
which Gallo used to "isolate" HIV), as well as other cell used for "HIV
isolation", CEM, C8166, EBV transformed B-cells and umbilical cord blood
lymphocytes.
8. HIV experts are unanimous that for infectivity, gp120 is
essential. Indeed, in 1988 your colleague JN Weber and you wrote, "The
first step in any viral infection is the binding of the virus particle
to a component of the host cell's membrane...For some time it has been
known that the binding takes place when CD4 interacts with an "envelope"
protein of the virus called gp120". However, Hans Gelderblom and his
colleagues at the Koch-Institute in Berlin, who have conducted the most
detailed electron-microscopy studies of "HIV particles" have shown that
the knobs on the surface of the particles, where the gp120 is found, are
only present in immature (budding) particles, which are "very rarely
observed". "Mature", cell-free particles do not have knobs, that is,
gp120.{Hausmann, 1987 #600 Thus it would impossible for Gallo to produce
"transmission by particles" by cell-free culture supernatants.
Surely, one would have to conclude, given the non specificity of
particles, reverse transcription (assumed to be due to reverse
transcriptase and no other enzyme activity) and antigen/antibody
reactions, Barré-Sinoussi's et al, Gallo's et al and Levy's et al data
do not prove the isolation of a retrovirus, even allowing your
definition.
When asked in 1997 by Djamel Tahi if Gallo purified HIV, Montagnier
replied "I don't believe so". He also added that when one uses leukemic
cell lives derived from patients with adult T-cell leukemia (as is the
case with the H9 cell line) which Gallo claims is caused by the
retrovirus HTLV-I and thus must be present in the culture, "You have a
mix of HIV and HTLV, it is a real soup". In other words, even if one has
proof for the isolation and the propagation of a retrovirus, it is not
proof that the retrovirus is HIV. It could very well be HTLV-I.
Montagnier forgot to mention that in the soup one could find at least
one more retrovirus, an endogenous retrovirus. The difference between
retroviruses and all other viruses is that while the finding of a virus
in a culture is proof that the virus has been introduced from outside
(the virus is exogenous), the finding of a retrovirus is not. You know
better than me that Varmus pointed out "one of the most striking
features that distinguish retroviruses from all other animal viruses is
the presence in the chromosomes of normal uninfected cells, of genomes
closely related to, or identical with those of infectious viruses". If
the finding of particles and RT activity in the "infected" cultures, and
proteins in the "infected" cells which react with sera from AIDS
patients is proof of HIV isolation, what does the detection of the same
phenomena in non-infected cultures and cells prove?
You also know, in fact you are among the many scientists who have
proven, that cultures of non-infected cells, sooner or later, begin to
release endogenous retroviruses. Their appearance can be accelerated and
the yield increased up to a million fold by stimulating the cultures
with mitogens, co-cultivation or by adding to the cultures supernatant
from normal, unstimulated cell cultures. Indeed, as far back as 1976
retrovirologists such as George Todaro recognised that "the failure to
isolate endogenous virus from certain species may reflect the
limitations of in vitro co-cultivation techniques". Montagnier et
al (as well as everybody else) employed at least two of the above
techniques. In fact, both Montagnier and Gallo concede that not one of
the phenomena which they claim prove the existence of HIV can be
detected unless the cultures are stimulated. In other words, while the
finding of a virus in culture is proof that the virus was introduced
from outside (infectious virus, exogenous virus ) the finding of a
retrovirus is not. Evidence exists that 70% of AIDS patients and those
at risk have antibodies to endogenous retrovirus. This means that even
if Barré-Sinoussi, et al Gallo et al and Levy et al, have all obtained
proof for the isolated (purification) of a retrovirus, even by your
definition, is not possible to claim that the virus is an exogenous
(infectious) virus. It could very well be endogenous and, as
retrovirologists have suggested, the result and not the cause of a
pathogenic process.
In 1997 in an article entitled "Current Problems and the Future of
Anti-retroviral Drug Trials", Joep Lange wrote: "There is also a
tendency to rapidly and widely publicise positive data and to delay or
refrain from publication of studies with negative outcome". In the case
of HIV, although both Montagnier's and Gallo's teams widely publicised
their claims of isolation (purification) of HIV, neither team published
electron micrographs of the material which in isopycnic sucrose
gradients bands at 1.16 gm/ml, the "purified HIV". At least in the case
of Montagnier's team (and according to Montagnier, Gallo's team as well)
this was because their results were negative. In scientific experiments
one manipulates the parameter whose effect is in question, executes
parallel control experiments with that parameter held constant, holds
other parameters constant throughout, replicates both the experimental
manipulation and the control experiment and obtains data. Given the fact
that retroviral phenomena are expected to arise spontaneously, let alone
be induced by the "experimental manipulation" absolutely essential to
obtain "HIV", in retrovirology controls constitute the quintessential
element. A common thread in "HIV" research is lack of controls. On the
rare occasions when authors do claim to use controls they are improper
and the experiments not performed blind. For example, "controls" consist
of cells cultured from healthy individuals, not sick individuals matched
to AIDS patients with respect to "other parameters". To date there is
only one exception and this from un-cultured tissue. In 1988 O'Hara and
his colleagues from Harvard reported "HIV particles" in 18/20 (90%) of
patients with enlarged lymph nodes attributed to AIDS and the identical
particles in 13/15 (87%) of patients with enlarged lymph nodes not
attributed to AIDS and at no risk for developing AIDS. These data led
the authors to conclude, "The presence of such particles does not, by
themselves indicate infection with HIV".
Molecular cloning cannot be considered proof for the discovery of a
retrovirus. To the contrary, cloning of the retroviral genome can only
be performed if and only if one first has a stretch of nucleic acid
which a priori has been proven to be the genome of a retrovirus. For
this one must first obtain particles in a purified form, or at least in
a material where there is not even a remote possibility of the presence
of an RNA apart from that contained in such particles. Then the
experimenter must show:
1. That the particles have all the morphological characteristics of
retroviruses;
2. That the particles are infectious;
3. Contain only RNA and not DNA;
4. The RNA codes for the particles' proteins.
To date there is not one single study proving even one of these
conditions, not to mention all of them.
Let us assume that HIV, unlike every other retrovirus or virus is, as
you say, a "virus without any proteins, particles etc.", that is, merely
a naked stretch of nucleic acid. If a stretch of RNA or DNA (the
provirus) is the genome of a unique retroviral particle then the most
basic requirement is proof for the existence of a unique molecular
entity. That is, all cultures and cells deemed infected must:
1. have a full length of the HIV genome, whatever that length is.
2. the stretch of DNA provirus, the genomes, must be identical.
To date the two "HIV DNA" or HIV DNA" of the same length have been
reported. Montagnier and his colleagues reported the "HIV DNA" to be 9 ±
1.5 Kb" whereas Gallo and his colleagues reported that "The overall
length of the HTLV-III provirus is approximately 10 kilobases". In the
1984 Levy and colleagues' first study of the "HIV genome", the "broad
band (> 15Kb) represents provirus integrated into host cell DNA". In
1995, Pasteur Institute researchers reported that "The complete
9193-nucleotide sequence of the probable causative agent of AIDS,
lymphadenopathy-associated virus (LAV) has been determined. The deduced
genetic structure is unique; it shows, in addition to the retroviral
gag, pol, and env genes, two novel open reading frames we call Q and F".
In the same year, Gallo and his colleagues reported their results on the
"HIV" nucleotide sequences using clone BH10 but also added, "The
sequence of the remaining 182 bp of the HTLV-III provirus not present in
clone BH10 (including a portion of R, V5, tRNA primer binding site and a
portion of the header sequence) was derived from clone HXB2...Of note is
the presence of a fifth open reading frame (nucleotides 8,344-8991)
designated 3' orf, present in clone BH8 but truncated in BH10". They
concluded, "The complete nucleotide" sequence of two human T-cell
leukaemia type III (HTLV-III) proviral DNAs each have four long open
reading frames, the first two corresponding to the gag and pol genes.
The fourth open reading frame encodes two functional polypeptides, a
large precursor of the major envelope glycoprotein and a smaller protein
derived from the 3' terminus long open reading frame analogous to the
long open reading frame (lor) product of HTLV-I and -II...The HTLV-III
provirus is 9,749 base pairs (bp) long" In 1990 the HIV genome was said
to consist of ten genes. In 1996 Montagnier reported that HIV possesses
eight genes and Barre-Sinoussi, HIV has nine genes.
Man and chimpanzee DNA differ by less than 2% but variation in the
composition of the "HIV genome" (derived from analysis of "pieces"
measuring 2% to 30% of the presumed total) measures between 3-40%. For
comparison, two RNA containing viruses (polio and influenza, the latter
after 27 years of dormancy,) vary by less than 1% as do RNA molecules
self-assembled in test tubes denied the organising influence of living
cells.
Given that the DNA sequence determines the composition of a virus's
proteins, and the latter the physical, biochemical and biological
properties of a virus, how is it possible for such variation to
represent one and the same agent? For example, how is it possible that
HIV can induce the same antibodies which can be recognised in a
universal antibody test containing identical proteins? Since Peter
Duesberg reminds us, "there is a range, a small range, in which you can
mutate around without too much penalty, but as soon as you exceed it you
are gone, and you are not HIV any longer, or a human any longer...then
you are either dead or you are a monkey, or what have you", it is
evident that whatever the "HIV DNA genome" represents, it cannot be a
virus.
If the stretch of "HIV RNA" is the genome of a unique exogenous virus
which infects individuals with AIDS or those at risk, then this RNA (or
cDNA) should be present in fresh uncultured tissue from all these
individuals and in nobody else. Furthermore, if in these individuals
there is massive HIV infection, as some of the best known HIV experts
claim, Southern/Northern blot hybridisation should be more than
sufficient to detect it.
I have always found it extremely difficult to understand why neither
Montagnier's nor Levy's groups reported such experiments in 1984 when
they claimed to have identified, characterised and cloned the HIV
genome. Gallo did and reported "HTLV-III DNA is usually not detected by
standard Southern blotting hybridisation ...when it is, the signals are
often faint...the observation that HTLV-III sequences are found rarely,
if at all, in peripheral blood mononuclear cells, bone marrow, and
spleen provides the first direct evidence that these tissues are not
heavily or widely infected with HTLV-III in either AIDS or ARC". These
studies were confirmed by many other researchers. The finding that when
the results were positive the hybridisation bands were "faint", "low
signal" was interpreted as proof that HIV seropositive individuals
contain HIV DNA in small numbers of cells and at low copy numbers, as
interpretation which became generally accepted, although Gallo and his
colleagues had an alternative explanation: "Theoretically, this low
signal intensity could also be explained by the presence of virus
distantly homologous to HTLV-III in these cells". This alternative
explanation has been ignored by everybody, including Gallo. However, at
a 1994 meeting held in Washington sponsored by the US National Institute
of Drug Abuse, Gallo admitted "We have never found HIV DNA in the tumor
cells of KS...In fact we have never found HIV DNA in T-cells". Data
which has come to light since 1984 suggest that Gallo's and his
colleagues' alternative explanation may be a fact:
1. At present there is ample evidence showing that normal human DNA
contains sequences related to HTLV-I and HTLV-II;
2. As late as 1994, writing in "Harrison’s Principles of Internal
Medicine", Gallo (and Fauci) taught medical students, "...there are no
known human endogenous retroviruses". This means that by "virus
distantly homologous to HTLV-III" they could have meant none other than
the exogenous retroviruses Gallo claimed to have discovered earlier,
that is, HTLV-I and HTLV-II. However, at present even Gallo admits that
the human endogenous proviral sequences "comprise about one percent of
the human genome";
3. Some of the best known HIV experts including Montagnier, Blattner
and Gelderblom agree that the pol and gag genes "may be highly conserved
between subtypes of virus". In a paper published in 1996 by Reinhart
Kurth and his colleagues one reads: "Retrotransposons evolved in a
variety of organisms ranging from protozoa to human beings. In these
elements, RT genes are linked to genes that code for polyproteins with
the potential to self aggregate and to form core particles. These
proteins are the equivalents of the retroviral capsid proteins usually
designated group-specific antigens (gag)...They [retrotransposons] may
be either the derivative or predecessors of retroviruses. Retroviruses
differ from retrotransposons by the presence of at least one additional
coding region, the envelope (env) gene". In 1984, Gallo group reported
that the "HIV genome" hybridised with the "structural genes (gag. pol,
and env) of both HTLV-I and HTLV-II. Obviously, the finding of a
positive hybridisation "signal" at least with an "HIV" gag or pol probe
is no proof for the existence of the "HIV genome".
In fact, at present evidence also exists which shows the presence of
"HIV" sequences in non-infected tissues. A .few examples suffice to
illustrate this point:
1. Although it is no longer accepted that HIV is transmitted by or is
present in insects, in 1986 researchers from the Pasteur Institute found
HIV DNA sequences in tsetse flies, black beetles and ant lions from
Zaire and the Central African Republic.
2. DNA extracted from thyroid glands from patients with Grave's
disease hybridised with "the entire gag p24 coding region" of HIV".
3. In 1986 you published a paper entitled "Isolation of a retrovirus
from two patients with "common variable hypogammoglobulinaemia" (CVH).
Patients with CVH "are prone to certain bacterial and mycoplasma
infectious but not to opportunistic viral protozal, and fungal
infectious, such as one seen in patients with genetic defects in
cellular immunity ..Both patients sera were negative for HTLV-III [HIV]
antibodies". I found your paper very interesting for two reasons.
Firstly, for what you meant by retroviral isolation and secondly, your
Southern blots results. By isolation you meant: "Extensive syncytium
formation was seen to polybrene-treated co-cultures, which on electron
microscopy showed a retrovirus morphologically indistinguishable from
HTLV-III/LAV" (see figure) and animal lentiviruses. Supernatant from
this co-culture was positive for reverse transcriptase, and the cells
were positive by immunofluorescence with serum from a patient with AIDS
and with the anti-HTLV-III monoclonal antibodies, a-p24 and a-a-p19
(from Dr R C Gallo). Southern blots of restricted DNA from infected
cells were probed with ABH-10 (from Dr R C Gallo ), indicated that the
viral genome showed homology to HTLV-III/LAV but with restriction enzyme
sites distinct from the prototype isolates, HTLV-IIIB and LAV. In other
words in 1986 by isolation you did not mean propagation but detection of
totally non-specific phenomena. More importantly, while Gallo et al
using ABH-10 as a probe could not find in T-cells from AIDs patients
"HIV DNA" you found it in the T-cells of patients with CVH.
In the second half of the 1980, in order to rescue the concept of an
"HIV genome" extensive use was made of the polymerase chain reaction.
However, the results with this technique were not much better than with
Southern blot hydridisation, and like Gallo in 1984, were interpreted as
proof that "HIV infected" individuals contain "HIV DNA" in a small
number of cells. A most striking feature of the results "is the scarcity
or apparent absence of viral DNA in a proportion of patients" and when
found, the "signal is very low". In addition, the specificity of the PCR
has never been determined. In the only study in which an attempt was
made, using antibody tests as a gold standard (everyone should know that
these cannot be used as a gold standard), it was found that in "Seven
French laboratories with extensive experience in PCR detection of HIV
DNA", the concordance between HIV serology and "HIV DNA" varies between
40-100%. Even if the PCR were specific, with this technique one detects
only a small portion of the genome.
In the vast majority of cases the presence of the "HIV genome" is
proven by amplifying short" invariant regions" of a "viral gene",
usually of the gag gene. However, since it is accepted that a
significant proportion of the "HIV genomes" are defective, finding a
fragment of a gene is not proof of the existence of the whole gene and
even less so for the existence of the whole genome "HIV DNA" or HIV
RNA", a fact accepted by many HIV/AIDS researchers.
In yet another effort to rescue the "HIV genome", tests have been
developed which claim to measure the "HIV RNA" in plasma which in turn
is said to quantify the number of particles in blood, the "viral load" .
One does not need to enumerate the many problems of these tests, a
glance at the following table suffices to realise that such tests do not
exist.
HIV-1 STRAIN |
RT-PCR |
BDNA |
NASBA |
DJ258 |
<400 |
111,500 |
100,000 |
DJ263 |
<400 |
79,800 |
60,000 |
SF2 |
225,500 |
38,000 |
240,000 |
III-B |
54,000 |
17,000 |
360,000 |
ZAM18 |
78,300 |
70,000 |
66,000 |
ZAM20 |
178,800 |
125,800 |
420,000 |
UG270 |
179,800 |
29,200 |
170,000 |
UG274 |
320,000 |
41,400 |
32,300 |
CM241 |
18,800 |
72,800 |
35,000 |
CM235 |
4,700 |
52,000 |
15,000 |
163.3069 |
36,200 |
94,000 |
57,000 |
162.307 |
2,800 |
78,100 |
26,000 |
G98 |
254,700 |
269,000 |
<400 |
LBV21 |
184,500 |
295,000 |
<400 |
VI557 |
950,000 |
587,000 |
125,000 |
The three assays frequently used to quantify the "viral load" are
reverse transcription-polymerase chain reaction (RT-PCR), nucleic acid
sequence-based amplification (NASBA) and branched chain DNA (bDNA). To
assess the impact of the assays used and of "genetic variability in
HIV-1 RNA quantification", researchers from France "evaluated three
commercial kits by using a panel of HIV-1 isolates representing glades A
to H...These isolates were expanded in culture. Virus was collected by
ultracentrifugation and resuspended in HIV-seronegative plasma. To
standardize the quantities of virus to similar levels in each
preparation, the p24 antigen was determined and the volume adjusted so
that each specimen contained approximately 10pg of p24 antigen per ml".
Since all the samples had the same quantity of "HIV", of "HIV RNA", one
will expect all the number in the table to be similar if not identical.
Looking at the rows the only conclusion one can draw is that at least
two of the assays do not measure "HIV RNA". Similarly, from the columns
the only conclusion one can draw is that either no one of the test
measure "HIV RNA", and/or there is no such thing as "HIV RNA".
Rich et al state, "Plasma viral load ["HIV" RNA] assays are designed
for monitoring the effectiveness of antiretroviral therapies and for
measuring the quantity of virus in patients with confirmed HIV
infection, not for the diagnosis of HIV infection. Their performance in
patients who are not infected with HIV is unknown" and their use leads
to "Misdiagnosis of HIV infection".
The fact that patients where "viral load decreases to undetectable
level still develop AIDS" also means that either the test do not measure
"HIV RNA" or the cause of AIDS is not HIV.
In summary, unless one draws conclusion "beyond the validity of the
data", it is not possible to conclude that "Barré-Sinoussi et al Gallo
and Levy" have proven the isolation of a unique retrovirus, even by your
definition of isolation. Neither do the present data prove the existence
of a unique molecular entity "HIV DNA", which constitutes the genome of
a unique retrovirus, much less of a retrovirus which infects humans.
To answer the Continuum challenge regarding proof for the isolation
of HIV, and thus its existence, Edward King from AIDS Treatment Update
presented you the challenge. In the April 1996 where your comments were
published one reads: "Gene cloning techniques allow researchers to
extract the viral genes found in HIV-infected cells. When the complete
set of genes is re-introduced into healthy human cells in culture, the
cells produce HIV particles". There is no proof for the existence of the
"complete set of genes" that is of "HIV DNA" even in one cell of one
AIDS patient. The small number of "complete set of genes" reported so
far, all of which differ significantly from each other, are from
cultured immortal cell lines. Neither is there one single study which
proves that, "When the complete set of genes is re-introduced into
healthy human cells in culture, the cells produce HIV particles".
QUESTION 5
My question was:
You state: "When you have evidence of infection in culture,
purification is not particularly important". These researchers did not
know that their cultures were infected. This is what they were
attempting to establish. Surely you don't claim that electron
micrographs of some budding forms on the cell surface or some cell-free
particles in the culture supernatant which do not even have all the
morphological characteristics of retroviruses, are proof of infection?
Are you further arguing that, without isolation, that is, purification,
a scientist can obtain "HIV" proteins and RNA?
Your reply was:
Yes, I do claim that visualization of HIV by electron microscopy was,
in 1983/84, an important component of the collective data on virus
isolation. Taken together with virus propagation, reverse transcriptase
activity and enrichment of particles by isopycnic gradients, it
convinced me that HIV is a retrovirus. Even more so, it was Montagnier's
electron micrographs published in April 1984 and previously shown at
Cold Spring Harbor Laboratory in September 1983 that convinced me that
HIV was probably a lentivirus among retroviruses, as they resembled
particles of equine infections anaemia virus - a lentivirus first
propagated by inoculating a filtrate too small for bacteria to pass
through into horses and donkeys and causing disease. So yes, I am
definitely arguing that, disregarding your meaning of 'purification',
but with my meaning of 'isolation', you can make quite large amounts of
HIV proteins and smaller amounts of RNA.
My response:
In AIDS Treatment Update one reads: "other scientists have
highlighted the irrelevance of this insistence on purity if the HIV
particles themselves are clearly present, for example, it's like saying
that it is impossible to identify a German Shepherd dog by its unique
appearance, if it happens to be surrounded by a pack of poodles".
However, in the HIV cultures in addition to the myriad of other things
one could see a "zoo" of particles, looking more or less like one or
another type of retrovirus particles but no one having all the
morphological characteristics of retroviruses.
The correct analogy for HIV is a person with no knowledge of German
shepherds who possesses an aerial photograph of a zoo, expects to see
dogs (retroviruses) but all he sees is many objects some of which look
like animals (viruses) and decides that one of the objects is a dog, in
fact a dog with unique composition and behaviour without first showing
the object is (a) an animal; (b) the animal is a dog; (c) the dog is
unique.
It is interesting that you now claim in addition to electron
micrographs, "virus propagation, reverse transcriptase activity and
enrichment of particles by isopycnic gradients" are needed to convince
you that "HIV is a retrovirus". However:
1. "Virus propagation" presupposes you already know you have a virus,
therefore virus propagation can not be used to prove the existence of a
virus.
2. You agree that reverse transcriptase activity is present in all
cells (see my response to Answer 4). I will only add here that a decade
before Barré-Sinoussi and Jean Claude Chermann claimed proof for the
existence of HIV, they wrote:
"Since similar polymerase activity [reverse transcriptase activity]
has been found in normal cells, may be mainly ascribed to the cellular
enzyme. At the same time Gallo was proving the existence of RT activity
in PHA stimulated but not in unstimulated normal, non infected cells. In
his interview with Djamel Tahi, Montagnier initially insisted that this
activity was "truly specific of retroviruses," but later he said that it
was only "characteristic" of retroviruses. Yet while my medical students
and residents, and presumably their teachers, have been taught by
retrovirologists that reverse transcription is specific to retroviruses,
retrovirologists such as Montagnier and Gallo know differently.
3. "enrichment of particles by isopycnic gradients". Neither
Barre-Sinoussi, nor Gallo and Levy presented proof for any "isopycnic
gradients" including the 1.16 g/ml being "enriched" or even having one
retrovirus-like particle. According to Montagnier, such evidence was not
published because they (and in his view, Gallo's group as well) could
not find any retrovirus like particles in their "isopycnic gradients".
This fact alone proves beyond reasonable doubt they did not have a
retrovirus and that the RNA and proteins found at the 1.16g/ml band as
well as RT activity or any other effect induced by the material in the
band have nothing to do with HIV or any other retrovirus. To claim
otherwise is no different from me, as a surgeon, claiming to have a
patient’s gallstones isolated, that is, removed an in my hands, merely
because I have proven he or she has abnormal levels of liver enzymes in
his or her blood. May I repeat, to date nobody has published proof for
the existence in the "HIV" cultures, not to mention the 1.16 g/ml band
showing particles having all the morphological characteristics of
retrovirus particles.
You seemed to have misunderstood my question. I did not ask you what
amount of "HIV" proteins or DNA you can make. I am solely interested in
quality. Not quantity. Allow me to put the question differently: As far
as I know, one can claim that a protein or an RNA fragment belongs to an
object, for example either a human being or a virus particle, only if
proof exists that they come from the object. Do you disagree? In the
"isolate" (be it the culture supernatant or the 1.16 gm/ml band)
obtained by your "isolation" method, there is proof for the existence of
a myriad of non-retroviral objects containing proteins and RNA, but no
object which has even the morphological characteristic of retroviruses
not to mention the physical characteristics such as their density. How
it is possible then to prove that some of the proteins and some of the
RNA which banded at 1.16g/ml or present in the culture supernatant of
were "HIV" proteins and "RNA" as Montagnier and Gallo did?
QUESTION 6
My question was:
In your view is it scientifically valid to say on the one hand, as
Montagnier did, that the 1.16g/ml "material" did not have even particles
with the "morphology typical of retroviruses, while on the other hand,
asserting that the proteins and RNA were those of a retrovirus, HIV?
Your answer was:
Barré-Sinoussi et al published electron micrographs of early budding
forms of virus only, that were not immediately identifiable as
retrovirus particles. By September 1983, Montagnier's electron
micrographs looked more typical.
My response:
In the very first electron micrograph (published in May 1983),
Barré-Sinoussi, Montagnier et al not only immediately identified some
particles as retrovirus particles but they stated that: "Electron
microscopy of the infected umbilical cord lymphocytes showed
characteristic immature particles with dense crescent (C-type) budding
at the plasma membrane...This virus is a typical type-C RNA tumor
virus". In 1984 Montagnier, Barre-Sinoussi et al reported that their
virus was "morphologically similar to D particles such as those found in
Mason-Pfizer virus or the virus recently isolated from simian AIDS." (By
1984 researchers from the primate research centres in the United States
claimed the existence of AIDS in monkeys and that the cause of AIDS was
a type-D retrovirus similar to the Mason-Pfizer virus, a typical type-D
retrovirus and suggested that the monkey AIDS and these retroviruses
could be helpful in the study of human AIDS and "HIV"). In the same
year, in yet another publication, Montagnier et al claimed that the
"HIV" particles had "morphology similar to that of equine infectious
anaemia virus (EIAV), and D type particles". The EIAV and the visna
virus are neither type C nor type D retroviruses but lentiviruses, that
is, viruses which have different morphology and said to induce diseases
long after infection. (By the time this paper was published it was
realised that patients who had a positive "HIV" antibody test did not
develop AIDS immediately, that is, there was a delay between the
positive test and the appearance of AIDS). It is most astonishing that
the one and the same virus is able to change genus from a typical type-C
to a typical type-D particle and then to a completely different
subfamily, namely a typical lentivirus, apparently at will and according
to non-morphological criteria. These taxonomical differences imply that
if HIV was a newly discovered mammal, it could have been either human,
or a gorilla or an orang-utan.
All this aside, you have not answered my question. Even if Montagnier
had electron micrographs typical of retroviruses, they were all from the
culture. As Montagnier acknowledged, no retroviruses were seen at the
1.16g/ml band. What I am interested in is how can anybody take some
proteins and RNA from the 1.16g/ml band where there are no proof of any
retrovirus-like particles and call them "HIV proteins" and "HIV
RNA"?
QUESTION 7
My question was:
What possible justification can there be for (a) using these proteins
as antigens in an antibody test to prove infection of millions of people
by a deadly virus? (b) using this RNA to prove not only infection but
also to quantify the viral load?
Your answer was:
There are many different tests for HIV-specific antibodies. Today's
commercial test kits are based on oligopeptides and on proteins
manufactured from cloned HIV DNA. No biological test for anything is
100% specific and 100% sensitive, but today's HIV tests are as good as
tests for any other human viral pathogen. Likewise, today's PCR primers
are highly specific and sensitive for the major strains of HIV in
developed countries. Some 'outlier' strains, especially in Gabon and
Cameroon are not picked up quite as sensitively and therefore estimates
of viral load with these 'outlier' infections should be interpreted
cautiously.
My response is:
Could it be there is no scientific justification for taking some
arbitrary RNA and proteins from a material in which there is no proof
for the existence of even retrovirus-like particles and calling these
"HIV RNA" and "HIV proteins"?
Above I have discussed the PCR and the "viral load tests". Here I
would like to add:
1. If the "viral load" test measures the number of HIV particles in
the blood and if there is a high level of such particles as it is
claimed, then it should be no problem to detect such particles using the
electron microscope. Yet to date, there is not even one electron
micrograph proving that such particles do exist in the blood. The only
electron micrographs published to date are from lymph nodes. In your
1993 review article in Science entitled "How does HIV cause AIDS?" you
state, "It has long been known from electron microscope and
immunofluoresence studies (24) that HIV is found in massive amounts in
the lymph nodes, even in the asymptomatic phase of infection". In ref.
24 you cite three papers, two by Tenner-Racz et al and one by Armstrong
and Horne (the latter authors are from the Royal Perth Hospital where I
work). In none of these studies is there evidence for "immunofluoresence
studies" of particles. The immunofluoresence studies of Tenner-Racz are
of lymph nodes. However, others report similar findings in patients who
suffer from a number of non-AIDS related diseases, and also healthy
individuals. In the first paper Tenner-Racz et al examined the lymph
nodes of 9 patients with persistent generalised lymphadenopathy and two
with Kaposi's sarcoma. In fig.2 where their results appear, there are
three electron micrographs. In the first two there is one "virus-like"
particle in each. The third shows one "Profile suggesting budding". In
the second paper there is one electron micrograph showing one particle.
It is interesting that without any proof the "virus-like" particles in
the first study become "retrovirus particles" and in fact "AIDS-related
virus" in the second, although even to a non-electron microscopist it is
obvious that the particles lack the appearances of the putative "HIV"
particle. In the Armstrong and Horne paper there are a few more
particles which are sometimes referred to as "virus-like" and at other
times retrovirus-like and whose "morphology conforms" to the type-C
retrovirus, not lentivirus as HIV is supposed to be. (Armstrong has
retired from the hospital but I contacted Horne who confirmed these were
indeed type-C particles). More importantly, particles with morphology
ascribed to "HIV" are found in the lymph nodes of non-AIDS patients.
2. The absolutely necessary, but not sufficient condition which one
must satisfy before using "PCR primers" in any test is to prove such
primers originate from a retrovirus-particle. Since no such proof exists
the only conclusion a scientist can draw is that nobody knows what a
positive "HIV PCR" is detecting. That this is the case is confirmed by
the test manufacturers themselves. For example, Roche states in their
package insert, "The AMPLICOR HIV-1 MONITOR Test is not intended to be
used as a screening test for HIV-1 or as a diagnostic test to confirm
the presence of HIV-1 infection" (Roche Diagnostics, Branchburg, New
Jersey, Art. 07 5623 7). HIV/AIDS experts agree. For example, Rich et al
state, "Plasma viral load ["HIV" RNA] assays are designed for monitoring
the effectiveness of antiretroviral therapies and for measuring the
quantity of virus in patients with confirmed HIV infection [positive
antibody test], not for the diagnosis of HIV infection. Their
performance in patients who are not infected with HIV is unknown" and
their use leads to "Misdiagnosis of HIV infection". Thus clinicians like
myself are left wondering how to reconcile statements such as "HIV RNA
(cDNA) is the RNA of a unique retrovirus HIV" and "today's PCR primers
are highly specific and sensitive" with statements such as HIV PCR is
"not for the diagnosis of HIV infection" because this will lead to
"Misdiagnosis of HIV infection". It does not make any sense that the PCR
results are confirmed by the antibody tests whose specificity has never
be determined, and vice versa that the specificity of the antibody tests
is confirmed by PCR whose specificity is unknown.
3. In my view the antibody tests are the most important aspect in
HIV/AIDS. This is because:
(i) you acknowledge that there are "naturally existing viruses"
(endogenous, non-infectious viruses) and there is a very high frequency
of recombination of the retroviral genomes. "HIV" has only been
"isolated" from cultures. As far back as 1988 the CDC researchers
pointed out that "the culture technique determines the ability of
infected cells to produce virus in vitro but does not necessarily
indicate the status of virus expression in vivo." This means that even
if one isolates a retrovirus from the culture, and even the isolation is
according to our terms, such finding is no proof for the existence of
this virus in vivo. The limited number of in vivo electron microscopy
studies, failed to prove even the existence of retrovirus particles in
AIDS patients, much less of a specific retrovirus. The vast volume of
genomic studies, failed to prove the existence in even one single AIDS
patient of the full-length "HIV genome". The existence of HIV in vivo
then rests with the antigen/antibody reactions.
(ii) the antibody tests are the only tests routinely used to prove
"HIV" infection.
(iii) the only way the proponents of the HIV theory of AIDS can
defend their position is to reassert the correlation between AIDS and a
positive antibody test. In your 1990 Nature publication with Harold
Jaffe where you "argued against Duesberg's view" you wrote, "The
evidence that HIV causes AIDS is epidemiological and virological, not
molecular...HIV is the singular common factor that is shared between
AIDS cases in gay men in San Franciso, well nourished young women in
Uganda, haemophiliacs in Japan and children in Romania orphanages".
However, correlation does not prove causation and any "correlation"
between "HIV" infection and AIDS is man made. Prior to 1987, one "HIV
specific" WB band was considered proof of HIV infection. However, since
15%-25% of healthy, no risk individuals have "HIV specific" WB bands, it
became necessary to redefine a positive WB by adding extra and selecting
particular bands, otherwise at least one in every seven people would be
diagnosed infected with HIV. (Notwithstanding, in the MACS, one band
remained proof of HIV infection in gay men until 1990). On the other
hand, although AIDS began to decline in 1987,this trend was countered by
the addition of more and more diseases and, most recently, mere
laboratory abnormalities to each revision (1985, 1987 and 1993) of the
first, 1982 CDC definition. The net effect of these changes was to
maintain a correlation between "HIV" antibodies and "AIDS" amongst the
"risk" groups while the risk of an HIV/AIDS diagnosis outside these
groups remained slight. This was further accentuated by avoiding testing
outside the risk groups. However, when such studies were performed, for
example, (a) amongst 89,547 anonymously tested blood specimens from 26
US hospital patients meticulously chosen to be at no risk of AIDS,
between 0.7% to 21.7% of men and 0-7.8% of women aged 25-44 years were
found to be HIV WB positive. (It is estimated that approximately 1% of
men are gay. Also, at the five hospitals with the highest rates of HIV
antibodies, one third of positive tests were in women. Yet men vastly
outnumber women as AIDS patients). (b) the US Consortium for Retrovirus
Serology Standardization reported that 127/1306 (10%) of individuals at
"low risk" for AIDS including "specimens from blood donor centers" had a
positive HIV antibody test by the "most stringent" US WB criteria (see
below). Thus the correlation between "HIV" antibodies and AIDS, which
experts accept as the only proof that HIV causes AIDS, is not a
statistic related to the natural, unbridled activity of a virus but is
instead a contrivance generated by mankind. Let me repeat, correlation
does not prove causation, and the artificiality of this particular
"correlation" severely compromises any scientific analysis.
Although the "antigens" used in the HIV antibody tests are said to
originate in a unique retrovirus HIV, it is not important if one has a
single or "many different tests" or what is used as antigen. Antigens
used in serological tests for syphilis and infectious mononucleosis do
not originate in the causative agents. However it is absolutely
necessary to have tests which are specific. That is, well before
introducing the test into clinical practice, the laboratory scientist
must prove that the tests are positive only in individuals infected. The
specificity of the "HIV" antibodies for proving HIV infection can be
determined only by using "HIV" isolation (purification) as a gold
standard. This has not been done and presently cannot been done because
nobody has purified "HIV". That this is the core, and that there is no
proof that "There are many different tests for HIV-specific antibodies"
is acknowledged by some of the best researchers in HIV testing and
HIV/AIDS such as Philip Mortimer and William Blattner. Even if one uses
your definition of isolation and purification it is still not possible
for anybody to have determined the specificity of the Western blot, the
antibody test used to "confirm" all other antibody tests. This is
because the WB is not standardised. The criteria used to define a
positive test varied with time (the US Food and Drug Administration have
had three so far) and vary between countries or even between
laboratories in the same country.
Space does not permit for me to cite the vast volume of data
regarding the non-specificity of the 'HIV antibody tests" but I am sure
you know where to find the relevant data. Because of this I will limit
the discussion to the basic problems associated with the definition of
the "HIV" antigens and antibodies. Barré-Sinoussi et al and Gallo et al
took the proteins which banded at 1.16g/ml, the "purified" virus and
reacted then with the sera from AIDS patients and those at risk. The
proteins which were found to most often react with some of the sera
sometimes were said to be "HIV" proteins and the reacting antibodies,
"HIV" antibodies. From such a reaction it is not possible to determine
the origin of one reactant, much less of both.
Let us consider a very ideal situation. Firstly, 75% of the material
which bands at 1.16g/ml is in the form of particles ("enriched") which
possess all the morphological characteristics of retroviruses. The
remaining 25% is cellular microvesicles and other cellular, bacterial
and viral constituents. Secondly, all of the many antibodies present in
AIDS patients and those at risk are "monospecific", that is, they react
with none other than the inducing antigen. A reaction between a protein
in the 1.16g/ml band and an antibody in the sera proves that the protein
is present in the individual and it is immunogenic. But it does not
prove the origin of that protein, that is, whether the protein is
retroviral, viral, bacterial or cellular. If we come closer to reality,
that is if you accept the proven fact that there are no such things as
"monospecific" antibodies, that all antibodies are "polyspecific", that
is, they cross-react, then, from such a reaction one cannot ever prove
that the protein is present in the individual or that the antibody was
directed against it. The reality, the truth, is that:
(a) Barré-Sinoussi et al and Gallo et al did not prove that their
1.16g/ml band contained even one single particle with the morphological
characteristics of retroviruses.
(b) The AIDS patients and those at risk have a plethora of antibodies
including auto-antibodies, all potentially cross-reactive. This means
that the most likely origin of the proteins which band at 1.16g/ml is
cellular or maybe bacterial/viral and the least probable retroviral. The
same is true for the antibodies. That both the proteins and the
antibodies are not retroviral has been proven by well known HIV
protagonists. Let us assume that although there is no proof that
Barré-Sinoussi's and Gallo's 1.16 g/ml band did contain retroviral
particles, it did contain some disembodied retroviral proteins. In this
case, if one compares the proteins which band at 1.16g/ml from the "HIV"
infected cultures, the "purified" virus, with the proteins which band at
the same density from non-infected cultures, the "mock" virus, we would
expect the "purified" virus to reveal its "extra" proteins.
In the well known Bess and co-authors 1997 Virology paper, the
authors had three HUT-78 (H9) cultures, two infected and one uninfected
control. The proteins from the banded material from all cultures
including the control (column A), which they called "mock virus", were
compared using electrophoresis. They stated that the only difference
between the three strips was that the infected strips contained major
bands of p24. But these same bands, although weaker, are present in the
"mock" virus protein strip whereas, to be HIV proteins, requires them to
be present exclusively in the "infected" strips. When asked for proof
that p24 etc in the strips B and C were HIV proteins their answer was
that the labels were added for the reader's convenience at the
suggestion of the reviewers. So Bess and his colleagues have shown that
the same proteins are present in the pure HIV and "mock" virus.
In their effort to develop a vaccine, and because humans cannot be
injected with either HIV or "mock" virus, Bess and his colleagues first
injected macaques with the "mock" virus. (This is culture fluids from
the uninfected H9 clone of the human HUT78 cell line "purified" as it
would be to obtain "HIV" or "SIV"). After the initial immunisation, the
animals were given boosters at 4, 8 and 12 weeks. At fourteen weeks, the
monkeys were challenged with intravenous SIV prepared from the same
human cells as "mock" virus and then monitored for seroconversion with
the SIV Western blot. According to the authors, the animal immunised
with "mock virus" "did not seroconvert to viral proteins after
intravenous challenge with SIV", and "These results are the first
demonstration that immunisation with purified cellular protein can
protect from virus infection...It has recently been suggested that
immunisation with alloantigens might serve as a vaccine to protect
against HIV infection. Our demonstration...support this concept".
The underlying principle of immunisation is its specificity. That is,
to protect against microbe 'X', the person or animal must be exposed to
material from 'X' in order that the immune system generates specific
antibodies. For example, immunisation with hepatitis vaccine does not
protect against poliomyelitis. Since monkeys immunised with proteins
derived from uninfected human cells are protected from infection with
'SIV' prepared from the same uninfected human cells, "mock" virus and
"real" SIV must be identical. If such "mock" virus and "SIV" are one and
the same we would expect that when "SIV" is prepared in antigenically
different cells, for example, monkey cells, there will not be
"protection". This is in fact what Bess and his colleagues proved in
another experiment. The only logical explanation of these data is that
they reflect immune responses to cellular proteins. Thus SIV proteins,
and by inference, HIV proteins, are nothing else but cellular
proteins.
All data presented to date is consistent with the "HIV" proteins
being cellular. Using "HIV" antibodies as probes, "HIV" proteins have
been identified in the tissues of persistently HIV negative, healthy
individuals including blood platelet and skin cells, thymus, tonsil and
brain. As a mark of the bewildering status of the HIV theory, at least
to this clinician, while HIV proteins could not be found in the
placentas of 75 HIV positive pregnant women, they could be found in the
placentas of 25 healthy, HIV negative women.
Detection of p24 has been also reported in organ transplant
recipients. In one kidney recipient (the donor was negative for p24
antigen) who, 3 days following transplantation developed fever,
weakness, myalgias, cough and diarrhoea, all "Bacteriological,
parasitological and virological samples remained negative [including HIV
PCR]. The only positive result was antigenaemia p24, positive with Abbot
antigen kits in very high titers of 1000pg/ml for polyclonal and 41pg/ml
for monoclonal assays. This antigenaemia was totally neutalizable with
Abbot antiserum anti-p24...2 months after transplantation, all assays
for p24-antigen became negative, without appearance of antibodies
against HIV. Five months after transplantation our patient remains
asymptomatic, renal function is excellent, p24 antigenaemia still
negative and HIV antibodies still negative". Using two kits, the Abbot
and Diagnostic Pasteur, in one study, p24 was detected transiently in
12/14 kidney recipients. Peak titres ranged from 850 to 200 000 pg/ml
7-27 days post- transplantation. Two heart and 5/7 bone marrow
recipients were also positive, although the titres were lower and ranged
from 140-750 pg/ml. Disappearance of p24 took longer in kidney
(approximately 6 months) than in bone-marrow (approximately 4-6 weeks)
recipients".
When by 1988 it was realised that by Gallo et al, and thus by your
definition of isolation, the frequency of isolation was low, the
definition was changed. Since then the detection in the cultures of
proteins (since antibodies cross-react, the protein(s) may be other then
p24) which react with a p24 antibody is considered proof for
"isolation". However, in 1992, Jorg Shupbach, the principal author of
the third and co-author of the fourth of the 1984 papers published by
Gallo's group on HIV isolation, reported that the whole blood cultures
of 49/60 (82%) of "presumably uninfected but serologically indeterminate
individuals and 5/5 seronegative blood donors were found positive for
p24".
The non-specificity of the p24 antigen test is so obvious that it is
accepted by no less an authority on HIV testing than Philip Mortimer and
his colleagues from the UK Public Health Laboratory Service, "Experience
has shown that neither HIV culture nor tests for p24 antigen are of much
value in diagnostic testing. They may be insensitive and/or
non-specific".
Interestingly, so far the only evidence of an animal model for AIDS
has been obtained by allogenic stimulation, a procedure which leads to
the appearance of "type C particles". Since individuals belonging to the
AIDS risk groups are repeatedly subjected to alloantigenic insult, one
would expect these individuals to have a positive antibody test, and not
be surprised if they developed AIDS, without ever coming in contact with
a retrovirus, HIV.
Given these data it is no wonder that biotechnology companies such as
Abbott Diagnostics include in their package insert the statement: "At
present there is no recognised standard for establishing the presence or
absence of HIV-1 antibody in human blood".
That antibody tests cannot be used to diagnose infection with a given
retrovirus was known before the AIDS era. In 1974 Hans Gelderblom and
his colleagues wrote, "While the virus envelope antigens are primarily
virus-strain specific, the bulk of internal proteins of the virion with
molecular weight (mw) between 10,000 d and 30,000 d are group-specific
(gs) for viruses originating in a given animal species (gs-spec.
antigens). The major protein constituent of mammalian C-type
oncornaviruses with a molecular weight in the range of 30,000 d was
found to possess, besides gs spec. antigen, an antigenic determinant
that is shared by C-type viruses of many mammalian species including
monkeys and was thus termed gs interspecies (gs-interspec.) antigen".
The fact that the proteins with molecular weights between 10,000-30,000
and the antibodies which react with them are not specific for
retroviruses has been amply demonstrated in the AIDS era. A few
examples:
- According to the AID vaccine Clinical Trials Group, "The presence
of p24 band was common among low-risk, uninfected volunteers and
complicated the interpretation of the Western blot test results".
- If the p24 band in the WB is considered the result of specific
reactivity, then approximately 30% of individuals who are transfused
with HIV negative blood become infected with HIV as a result.
- A 40 year old, male, HIV antigen negative, heterosexual donor of
Rh negative blood was given six 5ml injections of donated Rh positive
serum, administered at 4 day intervals. His "wife and child were
seronegative on HIV ELISA". The donor serum "was shown to be negative
on HIV antibody and antigen ELISA". "Blood taken after the first
immunization was shown to be negative on HIV antibody ELISA and
immunoblot assay. After the second immunization a weak signal on
ELISA, slightly above the cut-off level, was monitored. After the
third immunization the signal was strong and immunoblot revealed
distinct interaction with p17 and p55 proteins. An even stronger
signal was monitored after the fifth immunization. Interaction with
p17, p31, gp41, p55 and some other proteins was evident".
- 11/208 (5%) of healthy blood donors and 10/50 (20%) of patients
with measles, mumps, herpes simplex, dengue and other viral illnesses
had either a p24 or p18 band on the HIV Western blot test.
- The "HIV proteins (p17, p24)" appear in the blood of patients
(previously negative for all HIV markers) following "transfusions of
HIV-negative blood and UV-irradiation of the autoblood".
- In 1991 Kion and Hoffman injected non-HIV-infected mice with
T-lymphocytes from another strain of non-HIV-infected mice. The
recipient mice developed antibodies to the HIV gp120 and p24 proteins.
- In 1991, Strandstrom and colleagues reported that 72/144 (50%) of
dog blood samples "obtained from the Veterinary Medical Teaching
Hospital, University of California, Davis" tested in commercial
Western blot assays, "reacted with one or more HIV recombinant
proteins [gp120--21.5%, gp41--23%, p31--22%, p24--43%]".
- Last year it was reported that 35% of patients with primary
biliary cirrhosis, 39% of patients with other biliary disorders, 29%
of those with lupus, 60% of patients with hepatitis B, 35% of
hepatitis C, all non-HIV, non-AIDS diseases, have antibodies to the
"HIV" p24 "core" protein;
Yet up till the end of 1987, by which time the vast majority of
haemophiliacs, gay men and blood recipients who are tested, anybody who
was tested and found to have a p24 band on the WB was deemed to be
infected with a deadly virus and continued to believe and behave so even
after the criteria were changed. In 1987 former US Senator Lawton Chiles
of Florida told an AIDS conference how twenty two blood donors were
informed they were HIV infected on the basis of an ELISA test whereupon
seven then committed suicide.
Before the AIDS era there was proof that the antibodies which reacted
with "retroviral envelope proteins", like those which reacted with the
"retroviral core proteins" were also non-specific. On the 31st of
January 1975 Science published a paper by Gallo and Gallagher entitled:
"Type C RNA Tumor virus isolated from cultured Human Acute Myelogenic
Leukaemic Cells". This type-C RNA tumour virus, retrovirus, became known
as HL23V. The evidence for the isolation of HL23V surpassed that of HIV
in at least two aspects. Unlike for HIV, Gallo and Gallagher, (a)
reported the detection of reverse transcriptase activity in fresh,
uncultured leucocytes; and (b) published an electron micrograph of
retrovirus-like particles banding at a sucrose density of 1.16g/ml. This
study was followed by several papers in Nature, including one entitled
"Infective transmission and characterization of a C-type virus released
by cultured human myeloid leukaemic cells", with you as co-author. In
this study you reported that the HL23V "C-type particles are infectious.
The virus may be propagated to high titre in several cell types and a
quantitative bioassay is described. The production of substantial titres
of virus has enabled us to characterise the virus in detail and to
compare it with and show that it closely resembles the C-type SSV and
its associated virus (SSAV) previously isolated from a fibrosarcoma of a
woolly monkey. But our data on the relative plating efficiency on human
and 1283 marmoset cells, the inhibition of reverse transcriptase
activity by specific antisera and the nucleic acid lybridisation studies
suggest that, though closely related, the HL23V-1 virus and SSV are not
identical....one interesting fact is that the SSV(SSAV) viruses were
obtained from a woolly monkey that has been a pet and was therefore in
close contact with a human population...If the HL23V-1 virus and SSAV do
indeed have a common origin, it is plausible that the monkey was
infected by the human virus". Gallo, you and your colleagues concluded,
"Preliminary serological studies indicate that antibodies to the virus
are widespread in the human population. Moreover, a closely related
virus has been isolated several times from normal human embryo cells by
Panem et al., and Nooter et al. described elsewhere in this issue the
isolation of an SSV-related virus from human leukaemic cells. We should
bear in mind, however that isolation of the virus from leukaemic cells
does not necessarily mean that the virus is aetiologically related to
the leukaemia. Tests are currently in progress to determine whether the
virus has oncogenic potential in animals".
The results of your serological studies which indicated that
"antibodies to the virus are widespread in the human population", were
published in 1977, where you concluded: "The serological studies
presented here an by others provide indirect evidence that the
infectious mode of transmission remains a real possibility in humans,
and suggests that infection with an oncornavirus may be extremely
widespread, though perhaps not typically pathogenic."
In 1980 two research group one from the Laboratory of Cellular and
Molecular Biology, National Cancer Institute, and another from the
Laboratory of Viral Oncology, Memorial Sloan-Kettering Cancer Center
found that the antibodies were 'directed against carbohydrate
structures' and concluded that "The results are consistent with the idea
that the antibodies in question are elicited as a result of exposure to
many natural substances possessing widely cross-reacting antigens and
are not a result of widespread infection of man with replication
competent oncoviruses" (retroviruses). In 1981 Gallo accepted the
evidence that the antibodies which reacted with the presumed viral
proteins of HL23V were not so directed "but against the carbohydrate
moieties on the molecule that are introduced by the host cell as a
post-transcriptional event, and which are therefore cell-specific and
not virus-specific." This discovery was of such monumental significance
that today nobody, not even Gallo, considers HL23V as the first human
retrovirus. Or even a retrovirus. While the demise of the HL23V
antibodies and thus of HL23V was based on the findings of two research
group, that the antibodies were against carbohydrate moieties, as we
have pointed elsewhere, there are numerous studies which show this is
also the case with the "HIV" antibodies:
1. "One half of the molecular weight of gp120 is represented by
oligomannosidic oligosaccharides...Polyclonal antibodies to mannan from
yeast also recognize the carbohydrate structure of gp120 of the AIDS
virus";
2. "The immunochemical determinants of the antigenic factors of
Candida albicans display a high identity with the glycoprotein
(gp) 120 of HIV-1: they contain a(1®2) and a(1®3) linked mannose
terminal residues";
3. Antibodies to the mannans of Candida albicans "block
infection of H9 cells by HIV-1" as well as the binding of lectins to
gp120;
4. Recognition of gp120 by antibodies to a synthetic peptide of the
same antigen was "partially abolished if it was absorbed with the total
polysaccharide fraction of C. albicans" while the antigen
recognition by antibodies to "gp120 from human T cell lymphotrophic
virus type IIIB", "was totally blocked". From these data the authors
concluded: "These results indicate that mannan residues of C.
albicans can serve as antigens to raise neutralizing antibodies
against HIV infection";
5. "Normal human serum contains antibodies capable of recognizing the
carbohydrate moiety of HIV envelope glycoproteins...from 100ml of human
serum approximately 200ug of MBIgG was recovered [MBIgG=mannan-binding
IgG]...MBIgG bound to HIV envelope glycoproteins gp160, gp120 and
gp41";
6. Kashala, Essex and their colleagues have shown that antibodies to
carbohydrate containing antigens such as lipoarabinomannan and phenolic
glycolipid that constitute the cell wall of Mycobacterium leprae,
a bacterium which "shares several antigenic determinants with other
mycobacterial species" cause "significant cross-reactivities with HIV-1
pol and gag proteins". This led the authors to warn that
among leprosy patients and their contacts there is a "very high rate of
HIV-1 false-positive ELISA and WB results", that "ELISA and WB results
should be interpreted with caution when screening individuals infected
with M. tuberculosis or other mycobacterial species", and
furthermore that "ELISA and WB may not be sufficient for HIV diagnosis
in AIDS-endemic areas of Central Africa where the prevalence of
mycobacterial diseases is quite high";
7. Not only mycobacteria (M. leprae, M.
tuberculosis, M. avium-intracellulare) but also the
walls of all fungi (Candida albicans, Cryptococcus
neoformans, Coccidioides immitis, Histoplasma
capsulatum, including Pneumocystis carinii), contain
carbohydrate (mannans). One hundred per cent of AIDS patients (even
those with "No candida clinically") have C. albicans antibodies,
leading researchers from St. Bartholomews and St. Stephen's Hospitals to
state: "It is possible that candida may act as a cofactor in the
development of overt AIDS in HIV infected individuals". It may also be
of interest to note that in gay men the only sexual act which is a risk
factor for seroconversion is passive anal intercourse (exposure to
semen) and that mannose is present in both sperm and seminal plasma.
8. Since antibodies to mannans react with the "HIV proteins" then, as
Essex and his colleagues15 have pointed out for mycobacterial
infection in Africa, one would expect the sera of all people infected
with fungi and mycobacteria to cross-react with the "HIV-1
glycoproteins" as well as to cause "significant cross-reactivities with
HIV-1 pol and gag proteins".
9. Researchers from the University of Rome injected healthy mice with
an E. coli lipopolysaccharide (LPS) and reacted their sera with
two synthetic peptides, one encompassing gp120 V3 loop of "HIV-1 MN" and
the other "representing a gp41 immunodominant epitope". The "LPS-treated
mice showed a significant antibody reactivity" with the two peptides. (V
Colizzi et al., personal communication).
10. In the same study, the authors reported data from the sera of
HIV-negative subjects with autoimmune disorders. Recombinant gp120 and a
panel of synthetic peptides derived from the amino acid consensus
sequences of the HIV gp120, gp41, p24 or several unrelated proteins were
tested by specific ELISA. "The first set of experiments performed on
four patients with Sjorgern syndrome (SjS) and four patients with
systemic lupus erythematosus (SLE) revealed a significant anti-gp120
reactivity compared to healthy HIV-negative controls. Moreover, such
binding could be almost completely inhibited by preincubation with free
gp120. A significant anti-p24 reactivity was observed in 18 out of 29
[62%] sera from SjS patients and in 13 out of 25 [52%] from SLE
patients, while anti gp41 was observed only in 3 out of 14 [21%] SjS and
in 2 out of 20 [10%] SLE affected patients. Similar analyses were
performed in the murine model of autoimmunity, showing that sera from
MRL/lpr mice were able to bind all HIV related peptides in age-dependent
manner. The analysis of a panel of HIV unrelated peptides showed that
SLE as MRL/lpr sera bind both HIV related and unrelated peptides while
SjS sera failed to do so". In other words, sera which contain
autoantibodies react with the principal "HIV proteins" gp120, gp41 and
p24.
11. The same authors also reported similar results from (i)
experiments where "Two month old male CBA mice were immunized for 6
weeks with 50x106 allogenic lymphoid cells obtained from
either BALB/c or B6 male mice"; (ii) "Sera from 62 polytransfused (at
least 10 tranfusions/year) patients with thalassemia".
12. Similarly, in 1991, Kion and Hoffman reported "Mice of the
autoimmune strains MRL-lpr/lpr and MRL-+/+ made antibodies against
gp120". Mice that have been exposed to T-lymphocytes from another murine
strain were shown to make antibodies against gp120 and p24 of HIV.
These data pose the following crucial questions:
1. Given the fact that individuals with fungal and mycobacterial
infections have antibodies which may react with "HIV proteins" in the
absence of "HIV", and that E. coli is an intestinal commensal and
a potential bacterium in all of us, how can one assert that:
(a) reactions between antibodies in the sera of AIDS patients and
proteins present in cultures derived from the tissues of AIDS patients
is proof that the reacting proteins are constituents of a unique
retrovirus HIV and the antibodies are specific to these proteins?;
(b) PCP, candidiasis, cryptococcosis, coccidioidomycosis,
histoplasmosis, tuberculosis or Mycobacterium
avium-intracellulare disease, that is, the vast majority of the
opportunistic infections (88% of AIDS cases diagnosed between 1988 and
1992 had one or more fungal or mycobacterial infections) which signify
AIDS are caused by HIV on the basis of a positive antibody test?
(c) a positive antibody test in individuals with fungal and
mycobacterial infections proves HIV infection?
Since:
(a) mice and patients with autoimmune diseases (SjS and SLE) and AIDS
patients share many clinical and immunological (autoantibodies)
manifestations;
(b) patients polytransfused with allogenic blood and mice injected
with foreign cells and foreign proteins develop "HIV antibodies" but are
not infected with HIV; why should gay men, IV drug users and
haemophiliacs, who are all exposed to foreign cells and/or foreign
proteins, not also develop "HIV antibodies" and not be infected with
HIV?
In my view your proof that HIV exists depends on your belief in a
nexus between a series of non-specific phenomena (particles, reverse
transcription, antigen/antibody reactions and PCR). I contend this does
not constitute proof any more than claiming a cigarette smoking patient
with weight loss, fever, sweats and coughing up blood has to be
suffering from a hitherto unknown and undiscovered variety of lung
cancer. I further argue that the epidemiological data, far from
"proving" these data are due to the effects of a unique retrovirus, do
exactly the opposite. Indeed, as I suggested in my introduction, these
data prompt an urgent reappraisal of how and why such non-specific data
have been interpreted as proving the existence of HIV. The mistake that
has been made is the assumption that finding antibodies that react with
culture proteins proves (a) the proteins are constituents of a
retrovirus; (b) the antibodies specifically react with these proteins;
(c) isolation of a virus. That this is not the case follows from an
appreciation of basic science and was, for retrovirologists, the lesson
of the HL23V era. It is uncanny that, by changing a few dates, nouns and
adjectives in the papers of the HL23V era, it is possible to arrive in
the HIV era.
In conclusion I thank you for your contribution and cordially invite
you to continue the debate in the spirit Robert Oppenheimer who wrote,
"One thing we find throughout the house [of science]: there are no
locks; there are no shut doors; wherever we go there are signs and
usually the words of welcome. It is an open house, open to all comers".
I also must ask that you refrain from comparisons between my views on
HIV isolation and David Irving’s on the Holocaust. I find the comparison
not only irrelevant but distasteful.
Yours sincerely,
VF Turner MD
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ADDITIONAL DATA RE PROTEASE INHIBITORS/HAART
*According to one Australian expert, the success of the protease
inhibitors is "the nail in the coffin" of dissident theories. In this
regard, the findings of a recent study are interesting. "To assess the
long-term effects of highly active anti-retroviral therapy, we examined
3O4 anti-retroviral-experienced patients who were placed on HAART for a
period of 18 months. The baseline CD4 count was 385 X lO6/1
and HIV RNA level was 3.2 log10 copies/ml. At baseline, 39
percent were classified as asymptomatic, 33 percent were symptomatic and
28 percent had an AIDS defining illness. The HAART regimens included 3-5
anti-retroviral agents at least one of which was a protease inhibitor.
After 18 months, 14 percent of the population remained asymptomatic, 10
percent of which had an undetectable viral load. 39 percent were
symptomatic and 47 percent of the population had an AIDS defining
illness. The average CD4 count after 18 months on HAART was 301.79 X
106/1 and HIV RNA level of 3.2 log10
copies/ml.
Christina M. Ramirez and Michael S. Gottlieb.
California Institute of Technology. Long-term Highly Active
Anti-Retroviral Therapy in an Anti-Retroviral Experienced Population.
Keystone Symposia HIV Vaccine Development: Opportunities and Challenges
and AIDS Pathogenesis. January 7-13th, 1999, Keystone, Colorado.
www.newsfile.com
ADDENDUM I
LETTER REJECTED BY NATURE
NO REASON GIVEN
26/2/99
Dear Sir,
Please find a small contribution to your Correspondence columns. We
look forward to an answer to this vitally important question.
Yours sincerely,
VF Turner
Department of Emergency Medicine
Royal Perth
Hospital
Perth, Western Australia
In news and views (Nature 4th February), Robin Weiss and Richard
Wrangham stated "the retrovirus that is the main cause of AIDS has been
a puzzle ever since it was discovered by Barre-Sinoussi and here
colleagues in 1983".
The senior, most quoted author of the 1983 Barré -Sinoussi et al
paper (1) entitled: "Isolation of a T-Lymphotropic Retrovirus from a
Patient at Risk for Acquired Immune Deficiency Syndrome (AIDS)"1 is Luc
Montagnier. To isolate HIV, the authors used the long accepted method
for retroviral isolation, that is, banding, (ultracentrifugation) in
density gradients. They claimed to have shown that the density of
1.16g/ml, the sucrose density at which retroviral particles band,
contained "purified, labelled virus", that is, the 1.16g/ml band
contained nothing else but retrovirus particles isolated from everything
else.
In an interview published late last year,2 given to the French
journalist Djamel Tahi, Montagnier was asked why they did not publish
electron micrographs proving that the 1.16g/ml band contained isolated
HIV particles. Montagnier answered: No such proof was published,
because, even after a "Roman effort", at the density of 1.16g/ml they
could see no particles with "morphology typical of retroviruses". He
gave similar answers to repeated questions, including "I repeat, we did
not purify", that is, isolate HIV.
Given the important and highly significant differences in the
consequences of these contradictory claims, it is a matter of urgency
for the other 11 authors of the 1983 study, and especially Barré
-Sinoussi and her co-workers from the Pasteur Institute, to either
confirm or refute Montagnier's statements?
Eleni Papadopulos-Eleopulos1 Valendar F.Turner2 John M.
Papadimitriou3 David Causer1 Bruce Hedland-Thomas1 Barry Page1 Charles
Geshekter4
(1) Department of Medical Physics, Royal Perth Hospital, Perth,
Western Australia. email vturner@cyllene.uwa.edu.au (2) Department of
Emergency Medicine, Royal Perth Hospital, Perth, Western Australia; (3)
Department of Pathology, University of Western Australia. (4) Department
of History California State University, Chico Chico, California USA
REFERENCES
1. Barré-Sinoussi, F., et al. Science 220, 868-71 (1983).
2. Tahi, D. Continuum 5, 30-34 (1998).
The complete Continuum reference is: "Did Luc Montagnier discover
HIV? Text of video interview with Professor Luc Montagnier at the
Pasteur Institute July 18th 1997" http://www.virusmyth.com/aids/data/dtinterviewlm.htm
ADDENDUM II
REJECTED BY NATURE
NO REASON GIVEN
HIV TRANSMISSION FROM CHIMPANZEES
In their letter to Nature (February 4th), Gao et
al claimed to have proven;
(i) the identification in a chimpanzee of the genome of a new simian
immunodeficiency virus (SIVcpz), SIVcpz US;
(ii) that their chimpanzee, Marilyn, as well as two of the other 3
chimpanzees in which to date a SIVcpz was reported, GAB1 and GAB2,
belonged to a chimpanzee subspecies, P.t. troglodytes;
(iii) "the natural range of P.t. troglodytes coincides
uniquely with areas of HIV-1 group M, N and O endemicity";
(iv) "HIV-1 infection of humans occurred as a result of cross-species
transmission of SIVcpz from P.t. troglodytes".
(v) "P.t. troglodytes is the primary reservoir for HIV-1";
A close analysis of the evidence on which they are basing their
claims raises several questions:
(a) Marilyn was "wild-caught in Africa (country of origin unknown),
exported to the United States as an infant".
Two of the chimpanzees, GAB1 and GAB2, originated in Gabon. GAB1, who
was 4 years old when reported HIV-1 positive, was caught when she was
"about 6 months" and was kept with another 49 wild-caught animals at the
International Centre of Medical Research (CIRM) in Gabon. GAB2, who was
also reported HIV-1 positive, was about 2 years old when she was shot in
the wild, kept in a village for 2 days and then was brought to CIRM,
"where it died of its wounds one week later".
In the 1989 study where the "HIV-1 seropositivity" of GAB1 and GAB2
was reported, the authors (which included one of Gao's co-authors,
Peeters) concluded: "...on examination, none of the people caring for
the animals and none of those living in the village showed antibodies to
HIV/SIV. Furthermore, the region where the chimpanzee was captured is
known to have a low seroprevalence rate...It has been suggested that
human AIDS retroviruses originated from monkeys in Africa. However, this
study and other previous studies on SIV do not support this suggestion".
In other words, by the time when "HIV infection" and AIDS already
reached their peak in the US, Europe and Australia, the number of
individuals proven HIV seropositive in Gabon were few if any.
In a 1990 study published in Nature by researchers from CIRM
and the Pasteur Institute, including Wain-Hobson, where the authors
described "the molecular cloning and sequencing" of SIVcpzGAB1, one
reads: "In Gabon, only 2 out of 83 chimpanzees tested were seropositive,
indicating that SIVCPZ is not widely dispersed in this
region...Of more than 250 chimpanzees caught over the last 15-20 years
in West Africa, none were seropositive. This might explain the absence
of naturally infected chimpanzees in captivity in the United States as
virtually all are of West African origin".
How is it possible to espouse "prevalence in the natural host
"geographic coincidence", and "plausible routes of transmission" as
evidence to substantiate the claims that HIV-1 originated in P.t.
troglodytes and that this sub-species is the natural reservoir for
HIV-1?
(b) The three P.t. troglodytes, GAB1, GAB2 and Marilyn were
said to be infected with HIV-1/SIVcpz on the basis of an antibody
test.
However given that:
(i) as Philip Mortimer points out, "…it may be impossible to relate
an antibody response specifically to HIV-1 infection".
(ii) when the blood was collected none of the animals was perfectly
healthy although none had AIDS.
(iii) the only way to prove the specificity of an antibody test is to
use the virus isolation as a gold standard. Although no effort has been
spared, no SIVcpz could be isolated either from GAB2 or Marilyn (see
comments below for GAB1).
How is it possible to claim proof for infection on the basis of an
antibody test?
(c)
(i) If GAB1 and Marilyn were infected then, given that the animals
were brought to the colony as infants where no other animals or humans
working there were infected and, according to Weiss, "Chimpanzees in
captivity are mostly taken from the wild before they become sexually
active and so rarely harbour SIV", how did these two chimpanzees become
infected?
(ii) Since the three chimpanzees found positive were all female, and
since HIV/SIV is acquired following sexual maturity, how did they become
infected?
(iii) If the animals were infected with a virus SIVcpz and this was
transmitted to humans, why was this not transmitted to any other of the
49 animals at CIRM where GAB1 was kept or to the 93 animals in the
colony where Marilyn was kept, not even to her 6 living offspring or her
mates? (By the age of 26 she had a total of 14 pregnancies).
(d) The additional "lines of evidence" that Gao et al use to
substantiate transmission are based on genomic studies. Gao et al
claimed to have shown that "All HIV-1 strains known to infect man,
including HIV-1 groups M, N and O, are closely related to just one of
these SIVcpz lineages, that found in P.t. troglodytes". Indeed,
if all these HIV-1 and SIVcpz strains represented one and the same
virus, then their genomes will have to be "closely related". In fact
they should represent a unique molecular entity. Even in the genomes of
RNA viruses, including influenza, which are considered to be most
variable, a 1% sequence difference is considered to represent "extreme
variability". This is because small genetic differences lead to
significant phenotypic differences. For example the difference between
the human and the chimpanzee genome is less than 2%.
In the 1989 study of SIVcpzGAB1 Peeters et al wrote: "Nucleic
acid hybridisation experiments appear to indicate that the virus is
different from HIV-1 and HIV-2". In the 1990 Nature paper by
researchers from CIRM and the Pasteur Institute, including Wain-Hobson,
one reads: "Several regions of the chimpanzee sequences were more than
50% divergent with respect to HIV-1BRU. Some parts of the
gag gene were almost as varied as the hypervariable regions in
env...The vpu gene found only in the type 1 viruses was
particularly different (64% divergent to HIV-1BRU)…It is also
apparent that the SIVCPZ genome was not simply a more
diverged HIV-1 isolate…It is not possible to conclude that
SIVCPZ was the precursor to HIV-1, if indeed infection ever
passed in that direction. Even given this premise the vpu data
indicates that SIVCPZ was not the immediate precursor".
In a 1994 study of the SIVcpzGAB2, Peeters and his colleagues wrote:
"The genetic distance between SIVcpz-gab [SIVcpz GAB1) and
SIVcpz-gab2 is 14.1%. Genetic distances to the HIV-1
genotypes A, B and D strains are 13.7 to 16.3%, whereas distances to
group O HIV-1 strains are 15.4 to 18.5%". Contrary to Gao, in 1994
Peeters and his associates concluded: "On the basis of their respective
distances to each other and to the HIV-1 strains SIVcpz-gab
and SIVcpz-gab2 can be assigned as representative for two
distinct genetic lineages of HIV-1-related chimpanzee lentiviruses". By
1993 it was reported that "in the A-G HIV-1 genotypes the
intra-genotypic gag distances averaged 7%, whereas the inter-genotypic
distances averaged 14%...The maximum level of variability in gag is
still well below that observed for the envelope region of HIV-1".
The HIV-1 group O has "65% similarity to HIV-1 and 56% similarity to
HIV-2 consensus sequences. The env gene of MVP-5180 [HIV-1 group
O] had similarities to HIV-1 and HIV-2 of 53 and 49%
respectively...Comparison of the MVP-5180 amino acid sequences with that
of the Gabon chimpanzee virus showed similarities of 70, 78 and 53% in
the gag, pol and env genes, respectively".
As far as the genomic differences between HIV-1 group N, on the one
hand, and group M and O on the other is concerned, it suffices to quote
from the 1998 study where its existence was first reported. "Proviral
DNA amplification with several sets of HIV-1 group M and O primers was
attempted on pelleted end-cultured cells. Amplification was negative
with eight different group M env, gag or pol
primers and five group O env or gag primers". How is it
possible to claim proof for the existence of a unique molecular entity
which constitutes the genome of a unique retrovirus HIV-1/SIVcpz?
(e) The only way to prove that an RNA (and its cDNA) is the genome of
a retrovirus is to demonstrate that it comes from a retrovirus particle
and such RNA codes for its proteins. This can be done only by obtaining
the particles separate from everything else, purifying, isolating
them.
In the 1989 study where Peeters et al reported the isolation
of SIVcpzGAB1, stimulated peripheral blood lymphocytes "from healthy
human donors" were cocultured with the same type of cells from the
chimpanzees. Supernatant from the coculture was centrifugated for 10
mins at 400.000g. Detection in the pellet of reverse transcriptase
activity, using An(dT)12-18 as template primer was considered
proof for SIVcpzGAB1 isolation. Such a method for viral isolation is no
different from claiming that elevations in serum liver enzymes proves
the existence of gallstones and moreover, that the gallstones have been
isolated from the patient and are in the surgeon’s hands separate from
everything else. The SIVcpzGAB1 "genome" was obtained either by
hybridising the RNA present in the pellet (they presented no proof that
the pellet contained even retrovirus-like particles), where one would
expect to find ample cellular RNA, with probes "from
HIV-1oyi, a Gabonese HIV-1 strain", or from
"SIVCPZ-infected human lymphocytes" again using
HIV-1oyi as a probe. The "genome" thus obtained was compared
with the genome of HIV-1BRU.
We could find no details as to how the HIV-1oyi "genome"
was obtained. HIV-1BRU is the "HIV-1" which according to
Weiss was "discovered by Barre-Sinoussi and her colleagues in 1983". The
senior author of the 1983 Barre-Sinoussi study was Luc Montagnier.
However, in 1997 Montagnier not only acknowledged that they did not
isolate HIV-1BRU, but their "pure" virus from where they
chose some RNA and called HIV RNA, did not even contain particles with
"morphology typical of retroviruses".
The only evidence ever presented as proving the existence of the
SIVcpzGAB2 genome was reported by Peeters and his associates and is as
follows: "From this chimpanzee we have been unable this far to isolate a
lentivirus, but some of the primary peripheral blood mononuclear cells
(PBMCs) have remained available in a frozen state. To investigate the
genetic relationship to the SIVcpz-gab isolate [SIVcpzGAB1],
proviral DNA was extracted from these primary PBMCs [no mention is made
how it was possible to extract the proviral DNA from the chimpanzee
DNA], and a 280-base pair (bp) fragment of the pol gene was
amplified by a nested polymerase chain reaction (PCR). Subsequently PCR
fragments were cloned and sequenced". No mention is made as to how they
obtained the PCR primer. Gao et al used "consensus sequences" as
primers and the following method: "Here we used the polymerase chain
reaction (PCR) to amplify HIV- or SIV-related DNA sequences directly
from uncultured (frozen) spleen and lymph-node tissue obtained at
autopsy in order to characterise the infection responsible for Marilyn's
HIV-1 seropositivity. Amplification and sequence analysis of subgenomic
gag (508 base pairs (bp)) and pol (766 bp) fragments
revealed the presence of a virus related to, but distinct from, known
SIVcpz and HIV-1 strains. Because virus isolation from the autopsy
tissues was unsuccessful, we used PCR to amplify and sequence four
overlapping subgenomic fragments that together comprised a complete
proviral genome, which we termed SIVcpzUS".
However,
(i) The specificity of the PCR for HIV has never been proven. The
only way to obtain such proof is to use virus isolation as a gold
standard. Even if one accepts the claims for SIVcpzGAB1 isolation, it is
agreed that although no effort has been spared SIVcpz could not be
isolated from the other two animals. This means that the PCR results
obtained for the genomes of SIVcpzGAB2 and SIVcpzUS are false.
(ii) Even if they were specific for retroviruses; given that:
(a) The genome of all human beings and animals contain retroviral
proviruses i.e. genomes of the endogenous retroviruses.
(b) There are homologies between the genomes of different
retroviruses especially in the gag and pol genes. In fact
according to Montagnier and Wain-Hobson the gag and pol
genes "are generally conserved among retroviruses".
(c) In not one of the studies which claimed proof for the existence
of the SIVcpz genomes did the authors use controls.
(iii) How is it possible to claim that the sequences detected in the
DNA "of SIVcpz-infected human lymphocytes", the PBMCs of GAB2 and in
Marilyn's "spleen and lymph-node tissue" were those of an exogenous
retrovirus which is transmitted from one chimpanzee to another and from
chimpanzee to humans and not those of an endogenous retrovirus?
(iv) Since there is no proof that the three chimpanzees ever came in
contact with HIV-1 infected humans or animals or that they transmitted
such a virus to other humans or animals, is it not more "plausible" to
conclude that if these animals did harbour a retrovirus, the retrovirus
was endogenous?
In analysing the "SIVcpz" molecular biology one cannot help
reflecting upon the words of Sir John Maddox, "Is there a danger, in
molecular biology, that the accumulation of data will get so far ahead
of its assimilation into a conceptual framework that the data will
eventually prove an encumbrance? Part of the trouble is that excitement
of the chase leaves little time for reflection. And there are grants for
producing data, but hardly any for standing back in contemplation".
Eleni Papadopulos-Eleopulos1 Valendar
F.Turner2 John M. Papadimitriou3 David
Causer1 Bruce Hedland-Thomas1 Barry
Page1 Charles Geshekter4 Etienne
DeHarven5
1Department of Medical Physics, Royal Perth Hospital,
Perth, Western Australia. email
bruce.hedland-thomas@rph.health.wa.gov.au 2Department of Emergency
Medicine, Royal Perth Hospital, Perth, Western Australia; 3Department of
Pathology, University of Western Australia. 4Department of History
California State University, Chico Chico, California USA 5Professor
(Emeritus) of Pathology, University of Toronto, Toronto,
Canada.
REFERENCES
1. Gao, F., et al. Nature 397, 436-441 (1999).
2. Peeters, M., et al. Aids 6, 447-51 (1992).
3. Huet, T., Cheynier, R., Meyerhans, A., Roelants, G. &
Wain-Hobson, S. Nature 345, 356-9 (1990).
4. Mortimer, P.P. Med. Internat. 56, 2334-2339 (1989).
5. Weiss, R.A. & Wrangham, R.W. Nature 397, 385-386 (1999).
6. Gilden, R.V. et al. Lancet i, 678-679 (1986).
7. Steinhauer, D.A. & Holland, J.J. Annual Review of Microbiology
41, 409-433 (1987).
8. Janssens, W., et al. AIDS Research and Human Retroviruses 10,
1191-2 (1994).
9. Louwagie, J., et al. AIDS 7, 769-780 (1993).
10. Gurtler, L.G., et al. Journal of Virology 68, 1581-5 (1994).
11. Simon, F., et al. Nature Medicine 4, 1032-7 (1998).
12. Toplin, I. Spectra , 225-235 (1973).
13. Tahi, D. Continuum 5, 30-34 (1998).
14. Lower, R., Lower, J. & Kurth, R. Proceedings of the National
Academy of Sciences of the United States of America 93, 5177-5184
(1996).
15. Wain-Hobson, S., Alizon, M. & Montagnier, L. Nature 313, 743
(1985).
16. Maddox, J. Nature 335, 11 (1988).
Reference 13 is also available at http://www.virusmyth.com/aids/data/dtinterviewlm.htm
ADDENDUM III
19/7/99
Dear Mr Ranieri
I have been thinking about your e-mail of 8 April, and I realise that
I cannot have made myself clear when I first responded to your query. So
let me take up the two points you made, even though nearly 2 months have
elapsed.
1. You understand that Dr Turner and colleagues have not questioned
whether other viruses besides HIV exist, and cause disease. You are, of
course, absolutely right that a scientist does not prove that a
particular virus exists by pointing to the existence of others. That is
the inverse of the point I was trying to make. It is precisely because
Val Turner and his colleagues in Perth have not queried the existence of
other viruses that I find it difficult to take their ideas on HIV
seriously. All the 'failings' they attirbute to HIV could equally well,
according to their own stringent criteria, be levelled against any virus
with a lipid envlope, eg small pox, influenza, measles, mumps or yellow
fever. The evidence is as good, or rather, in Dr Turner's eyes as bad,
as for HIV. No virologist in the 101 year history of the discovery of
viruses ever demanded for any virus the definition of 'purification'
that they and the magazine Continuum claim are the standard criteria of
virus isolation. So we end up talking at cross-purposes.
2. You find it difficult to accept that I can bow out of this
discussion. Let me explain by analogy. Do you expect today's physicists
to spend time in prolonged e-mail 'discussion' with members of the flat
earth society? Or do you think modern astronomers are merely speculating
to hold, as Copernicus, Keppler and Galileo bravely did, that the
planets orbit around the sun rather than the earth? Would you think it
irresponsible for an astronomer in 1999 to bow out of such a discussion
if a believer in a flat, planar universe was ignored and then claimed
his view was deliberately suppressed by the scientific orthodoxy of the
day? So who do you think is the Galileo on HIV - Turner or
Montagnier?
Now let's come back to my argument under item 1, that the evidence
the existence of viruses like measles or yellow fever is no better than
that for HIV. Even if as a non-scientific journalist you did not wish to
take sides, would you think it logical to hold that Jupiter, say, orbits
the earth while Mars, Venus, Saturn, etc, orbit the sun?
As to bowing out, perhaps I should not have bowed in my replying to
Dr Turner in the first place. Dr Turner e-mailed me after I co-authored
a short commentary in Nature on SIV in chipanzees being a possible
source of human HIV, based on other scientists' work. He contacted me,
to clarify my views on the existence of HIV and on a quote of
Montagnier, because his patients were anxious about it (do surgeons
treat AIDS?). After he said he passed his reply to you (are you his
patient?), [Ranieri is not a patient] he told me that any communication
he receives from me is his property. I now realise that Dr Turner was
spoiling for a fight. He knew my views already, but wished to engage in
argument in a manner that to me seems strange to open, cordial
scientific dispute. So I chose to bow out.
However, it would be difficult to accuse me of supposing the debate
about whether HIV causes AIDS. Together with Dr Harold Jaffe I have
stated my views succinctly and cogently nine years ago (Nature 345:
659-660, 1990). I have nothing more to add, or subtract, from what we
put on record then, except that what new, reliable data have come to
light since then confirm our views [Duesberg never took anyone to task
over the isolation/existence of HIV. The Perth group fare still the only
scientists to publish these arguments]. So why should I go on repeating
myself? If I were a radio or television producer, I think I might seek
to explore a programme on latter day flat-earthers as a sociological
phenomenon. What lies behind HIV and AIDS denial? Why do such people
feel persecuted by scientific orthodoxy when in reality they are simply
ignored? Why do quasi-fascist concepts keep slipping in, like the holy
grail of 'purification'? Doesn't HIV denial resemble holocaust denial?
First, I would take a vacation on the beach with the writings of two
Italian Nobel Laureates who had suffered under fascism: Enrico Fermi the
physicist who discovered nuclear fusion, and Primo Levi, the chemist who
won the Nobel Prize for Literature, but later took his own life. These
two thinkers would help me to place the issues of scientific controversy
and society in proportion. [Why not Giordano Bruno?]
If all that seemed too strong to my director of programmes for family
viewing, I would propose a documentary to mark the centenary of the
discovery of viruses, ie distinguishing them from other microbes. How
influenza killed more people in 1918/19 than all who died in the Great
War, whether it will happen again, why the Hong Kong chicken flu didn't
spread. How WHO conquered smallpox, controlled yellow fever and is on
its way to eradicating polio. Why HIV, cancer viruses and herpes viruses
cannot be eliminated in the same way. Where HIV came from and where it
may be going. Will new viruses appear? Should we be scared about Ebola
and Lassa fever spreading to Western Australia? Why did a virus related
to measles transfer from fruit-bats to horses and thence to humans in
Queensland three years ago, and in Malaysia three months ago. Are the
fruit-bats in the surburbs of Australian cities therefore a threat to
human health? There's plenty of good material for responsible,
entertaining and lively debate about viruses waiting for an enterprising
reporter.
Kind Regards
Robin A Weiss
ADDENDUM IV
Postscript March 13th 2002. Since Professor Weiss' comments I have read a few of Primo Levi's books myself. I found this prescient quote in one of them:
In countries and epochs in which communication is impeded, all other liberties soon wither; discussion dies by inanition, ignorance of the opinion of others becomes rampant, imposed opinions triumph; (the well known example of this is the crazy genetics preached in the USSR by Lysenko, which in the absence of discussions (his opponents were exiled to Siberia) compromised the harvests for twenty years. Intolerance is inclined to censor, and censorship increases the ignorance of the arguments of others and thus intolerance itself: a vicious circle that is hard to break.
Primo Levi The Drowned and the Saved 1988
(1) Dr. VF Turner Consultant Emergency Physician Royal
Perth Hospital Perth, Western Australia Voice +618
92242693 Fax +618 92247045 Email vturner@westnet.com.au |
(2) Robin A Weiss Professor of Viral Oncology Wohl Virion
Centre Windeyer Institute of Medical Sciences University
College London 46 Cleveland Street London W1P 6DB,
UK Tel: +44 207 679 9554 Fax: +44 207 679 9555 email: r.weiss@ucl.ac.uk |