Professor William Makgoba
sent the email below to the
MAKGOBA TO THE PERTH GROUP
I write to you to confirm that the preadsorption experiments or studies are part of the protocol that we fully endorse from the CDC, MRC, Harvey and yourselves. In the interest of efficiency and taking advantage of modern science I wish to suggest that we drop the issue of HIV isolation as you propose for the following reasons: the HIV has been fully sequenced and shown to have a unique genetic sequence that falls within the retroviral family; almost every lab in the world uses PCR and transfection experiments rather than sucrose density gradients for the isolation and culture of the virus. While this technology might have been useful 16 years ago, science and technology has moved on and every South African lab uses PRC and culture for viral isolation, culture and sequencing. There is no more definitive characterisation of an agent than DNA sequencing today. My plea would be to stick to the adsorption studies and combine them with the molecular beacon studies. From the results of these two we should reflect on the best way forward together. I hope you will find my request acceptable.
Looking forward to your response.
PERTH GROUP TO MAKGOBA
Thank you once again for your email of
We would also like to propose that you
and your colleagues contact Djamel Tahi in
Djamel Tahi email <email@example.com>
Telephone int + 336 624 90117
Professor Sam Mhlongo has a copy in French with transcripts in French and English.
RESPONSE TO THE REQUEST BY PROFESSOR WILLIAM MAKGOBA TO DELETE VIRUS ISOLATION FROM THE PRESIDENTIAL PANEL EXPERIMENTS
First, to avoid any misinterpretations, it may be advantageous to define and discuss the terms isolation, purification and transfection.
Isolation (Latin insulatus = made into an island), means the act of separating an object from all other material of any consequence. However, it is apparent that virologists do not use the term in this sense. Rather virologists regard isolation as the detection of one or several phenomena which they assert singly or collectively are virus specific and hence proof for the existence of a particular virus. For example, Barré-Sinoussi, Chermann and Montagnier detected reverse transcription of the template-primer An.dT15 in a culture of lymphocytes from a patient at risk of AIDS. This was interpreted by them as proof for the isolation of a virus. The detection of the same phenomenon in a sub-culture was considered proof for propagation of the same virus.
Purification means to obtain an object free from contamination. In the dictionary sense isolation = purification. It is highly significant that while purification may or may not play a role in the isolation of a particular virus (virological sense), purification is a completely separate and pivotal matter in defining which biochemical constituents are viral and which are not. For example, Montagnier and his colleagues claimed to have proven their “isolate” was a retrovirus because they detected reverse transcriptase (RT) activity in the 1.16 gm/ml portion of a sucrose density gradient and that this band was “purified, labeled virus”(1).
Transfection means the introduction of exogenous DNA into cells and its ability to replicate and express itself in these cells, that is, transcription of DNA into RNA and translation of RNA into proteins. With the present day technology any fragment of DNA can be transfected. However, transfection does not prove a DNA fragment has a viral or cellular origin.
Our point by point response to your proposal is as follows:
1. The isolation experiment, like the preadsorption experiments, “we fully endorse from the CDC, MRC, Harvey ". In fact at the press conference it was repeatedly stressed that we should start from basics, and there is nothing more basic than HIV isolation.
1. "HIV has been fully sequenced and shown to have a unique genetic sequence that falls within the retroviral family”.
(a) The first absolutely necessary but not sufficient step to "fully" sequence the genome of a retrovirus and prove it has a "unique genetic sequence" is to obtain the RNA fragment from a retroviral particle. Since the retroviral particle measures 100-120 nM in diameter the next best thing is to obtain the RNA from a mass of material which contains nothing else (or at least no other biological material) but particles with the morphology of retroviruses. Even in the 1950s, as the well known retrovirologist JW Beard stressed, it was known that to define a nucleic acid or protein as viral, the following steps are absolutely necessary; "(2) recovery (purification) of the particles in a given preparation that are homogenous with respect to particle kind; (3) identification of the particles and (4) analysis and characterisation of the particles for the physical, chemical [RNA, proteins], or biological properties desired". He also showed that even then there were shortcomings in the "identification, characterisation, and analysis" of the particles, because "the need for the critical fundamental analyses that are dependent on access to homologous material", was ignored by some researchers (2). In the early 1970s it was Barré-Sinoussi and Chermann who showed the need for obtaining material which contained nothing else but particles with “no apparent differences in physical appearances” (3, 4). Presently, the view that the characterisation of a protein or an RNA fragment as retroviral is absolutely contingent upon purification of the retroviral particles is not only ours but also that of such eminent HIV experts as Luc Montagnier, Robert Gallo, Julian Bess, Larry Arthur, Hans Gelderblom and Pablo Glushankof (5-13). Unfortunately, as most of the above mentioned scientists have acknowledged (Robert Gallo may be an exception), “access to homogenous material” has so far not been achieved.
(b) To date no one has proven that the “HIV” genome has unique genetic sequences. In fact the data is that such "genomes" vary between 3-40%. On the other hand, variations beyond 1% in the genomes of RNA viruses such as polio and influenza viruses are considered to represent "extreme variability" (14).
(c) To date there is not one single study proving the existence of the “HIV” genome even in one single cell of even one single AIDS patient. All “HIV” sequences are obtained from cell cultures of immortalised cell lines.
(d) If one assumes there is a unique “retroviral family” of genomes, finding such a sequence in a cell of an AIDS patient is not proof that it is the genome of a unique retrovirus, HIV. This is because, with perhaps the exceptions of Robert Gallo and Anthony Fauci, (15) all retrovirologists accept that all cells possess genomes of the “retroviral family”. In fact, 1% of our genome is considered to be constituted of such sequences (16, 17) (http://www.virusmyth.com/aids/data/epreplypd.htm).
3. "almost every lab in the world uses PCR and transfection experiments rather than sucrose density gradient for the isolation and culture of the virus".
(a) At present one can transfect any fragment of nucleic acid. The question is not whether nucleic acid can be transfected but what is the origin of that nucleic acid? The object of interest to us all is a retrovirus, that is, a particle of defined taxonomy composed of proteins, RNA and lipids. A retrovirus is not merely a fragment of RNA (DNA). At present there is not even one single study to prove that transfection of the “HIV DNA” leads to the appearance of HIV particles in the culture.
(b) PCR can be used to detect and hence prove infection with HIV and then if, and only if:
(i) there is proof that the primers and probes originate from a mass of unique, purified retroviral-like particles proven infectious. Although, the ultimate origin of all the primers and probes employed in the "HIV PCR" test is sucrose density gradient banded material, to date no proof exists that the bands are purified retroviral particles. However, there is ample proof that the banded material, at best, consists of cellular fragments containing both RNA and DNA (5, 6). In the material from which Luc Montagnier and colleagues obtained their “HIV” nucleic acid they could not find even particles with “the morphology typical of retroviruses”.(18). The analogy we use when writing for non-scientists is that of a paternity suit. Proof of origin of primers and probes is no different from that which courts of law require to resolve disputed paternity. The child’s and the alleged father’s DNA are defined solely on the basis that specimens are taken from their respective bodies. If the HIV PCR test were a paternity suit the judge would pronounce the father guilty without proof that the DNA tested against that of the child came from the father or even a human being.
(ii) There is proof that the PCR specifically amplifies the sequences of interest and nothing else. This is not the case with the "HIV" PCR (19-21). In fact in the view of some retrovirologists, one should "NOT use" PCR to confirm HIV infection; (22) "the false-positive and false-negative rates of PCR that we determined are too high to warrant a broader role for PCR in either routine screening or in the confirmation of diagnosis of HIV infection" (21). According to researchers from the Massachusetts School of Medicine “Plasma viral [RNA] load tests were neither developed nor evaluated 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” (23). Manufacturer Roche state the following in their packet insert: “The Amplicor HIV-1 [RNA] Monitor test is not intended to be used as a screening test for HIV or as a diagnosis test to confirm the presence of HIV infection” (Roche Diagnostic Systems, 06/96, 13-08088-001. Packet Insert).
4. "While this technology might have been useful 16 years ago, science and technology has moved on and every South African lab uses PCR and culture for viral isolation".
(a) As long as one is able to obtain purified particles it is not important what "science and technology" one uses. However, today, like 16 years ago, there are only two methods to isolate (purify) retroviral particles: banding in density gradients and filtration, the former being accepted by nearly all retrovirologists as being preferable;
(b) The reaction between antibodies directed against a protein, p24 and antigens present in cultures is considered by "every South African lab" as proving HIV isolation. Under no circumstance can a reaction between antibodies to a protein and culture antigens (which may not be the same protein or indeed any protein) be considered proof for isolation or purification of a particle said to be composed of at least ten proteins, RNA and lipids. Such a reaction can be used only to prove the detection of HIV in a culture and then, if and only if there is proof that:
(i) the p24 protein is an HIV protein. Luc Montagnier was the first to claim proof that p24 is an HIV protein, a claim accepted by nearly everybody. According to Montagnier, "analysis of the proteins of the virus demands mass production and purification. It is necessary to do that" .(18). (It goes without saying that this step is also fundamental to obtaining the HIV RNA). Donald Francis, a researcher who with Gallo played a significant role in developing the HIV theory of AIDS, agrees. "The problem here is that such methods [antibody and nucleic acid tests] can be developed only if we know what we are looking for. That is, if we are looking for a known virus we can vaccinate a guinea pig, for example, with pure virus...Obviously, though, if we don't know what virus we are searching for and we are thus unable to raise antibodies in guinea pigs, it is difficult to use these methods...we would be looking for something that might or might not be there using techniques that might or might not work" (24) (italics ours). However, in July 1997 Montagnier admitted that he and his colleagues did not purify HIV. In the material from which p24 originated Montagnier and his colleagues could not find even particles with the morphology of retroviruses (18).
(ii) the antibodies to p24 react only with p24 and nothing else in the culture. However, the non-specificity of the p24 antigen test is obvious and 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" (25). Similarly, Jörg Schüpbach, principle author of one (9) of the four Science, May 1984 papers published by the Gallo group, has published data using detection of p24 to “isolate” HIV from cultures of whole unfractionated blood. He reported positive results in 49/60 (82%) of "presumably uninfected, but serologically indeterminate" individuals and in 5/5 "seronegative blood donors" (26). When Djamel Tahi proposed that “an antibody reaction with p24 is considered non specific”, Montagnier replied “It is not sufficient for diagnosing HIV infection…No protein is sufficient” (18).
In 1983 Montagnier and his colleagues claimed to have
proven the existence of HIV by obtaining "purified labeled
virus”, from a patient BRU (1). In 1997, in
response to the question “Why no purification?”, he
stated: "I repeat we did not
purify" (18). In 1997 when Montagnier was asked if Gallo achieved
the purification that Montagnier asserted necessary to define the HIV proteins
he replied: "Gallo? I don't know if he really purified. I don't believe so". And even if Gallo did purify, because he used
an immortalised cell line, which was infected with HTLV-I, (27, 28) the purified material would contain a mixture of
retroviral particles. Or
as Montagnier described it, "a real soup". On the other hand, as far as Gallo is
concerned, Montagnier's data did not prove "true isolation" (7). In a review (29) of Luc Montagnier's book, “Virus”, the eminent HIV expert Jaap Goudsmit wrote:
"While early independent isolations were made by Montagnier, Gallo,
and Levy in the years 1983 and 1984, Montagnier, Chermann and Barré-Sinoussi, were without any doubt the first to
identify the virus that was later shown by epidemiological research to be the
cause of AIDS". In his own
book: "Viral Sex" published in 1997, Goudsmit
wrote: "The BRU lymph node was
first cultured in early January 1983 and, on January 15, it shed an enzyme absolutely unique to the lentivirus group. The BRU virus grew slowly and with difficulty,
but its identity and activity were reported in the
One must also add that:
(a) Reverse transcriptase (RT), the “enzyme absolutely unique to the lentivirus group” is not even specific for retroviruses. RT is found in all cells including bacteria (31, 32) and hepatitis B virus. Nowadays, knowledge of its presence in the latter virus (which also infects T-lymphocytes (33)) is even available in the popular press to readers contemplating investment in biotechnology companies (34).
(b) HIV researchers do not distinguish between reverse transcriptase (the enzyme) and reverse transcription (the process). The latter is also nonspecific. For example, all three cellular DNA polymerases reverse transcribe. In fact, in 1975, an International Conference on Eukaryotic DNA polymerases defined DNA polymerase γ [gamma] as the cellular enzyme which "copies An.dT15 with high efficiency but does not copy DNA well" (35). An.dT15 is the synthetic primer-template in universal use to prove the presence of HIV RT and in some cases to prove “isolation” of HIV (36).
(c) Mitogenically, normal, non-HIV-infected human lymphocytes (37) as well as spermatozoa (38) reverse transcribe. Mitogenic stimulation of cell cultures is an absolute requirement in all HIV research to prove HIV “isolation” (39).
(d) The description Montagnier gave to BRU’s virus which he, Montagnier claimed to have isolated, was not a “lentivirus” but “a typical type-C RNA tumor virus”, a taxonomically distinct retrovirus which is not a Lentivirus (1).
1. The results of the antibody and PCR tests affect millions of people around the world.
2. It is absolutely necessary although not sufficient to obtain material from affected patients which contains nothing but particles with the morphological characteristics of retroviruses. This is the only way to characterise the proteins used in the antibody test as well as the primers and probes in the PCR tests as being HIV.
3. To date nobody has published evidence proving the purification of particles with the morphological characteristics of Lentiviruses (or even retroviruses) from cultures of fresh tissues from AIDS patients.
Then either HIV does not exist or the experiments need to be repeated and properly controlled. Therefore the experiments we have outlined in our proposal which directly address the problem should be performed.
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MAKGOBA TO THE PERTH GROUP
Thank you for the response. I cannot accept the arguments as framed. PCR is highly specific. DNA sequences have been compared through out the world for homology. Retroviruses have unique sequences perculiar to them. There is abundant evidence for this. There is no need or merit to focus on an interview that has been distorted by people. Some of us have spoken and visited Luc's lab to confirm for ourselves that indeed such a virus has been isolated and can be identified by its unique sequence. Perhaps the Perth group should do the experiment and prove every one wrong rather than entrench yourself in an untenable scientific position that is
against the best scientific data. For myself, I cannot see any South African scientist who would accede to your proposal. You may have to prove it yourselves. The panel did not accept this proposal. Peter Duesberg, an eminent retrovirologist has identified and sequenced an HIV. [We contacted Peter Duesberg who denied he has performed any such experiment]. Perhaps it might be better to address this matter with him, rather than keep playing the Luc Montagnier record. It was my hope that we should start from "doable studies"; build trust over time amongst each other and move to the difficult issues. This was my proposal. It is for me better to start from somewhere than to have the whole thing collapse and not being done. In that way you gain nothing out of this excercise. Our country will continue to work on the evidence that HIV causes AIDS. To turn this around requires not stubbornness but pragmatism that is built by small increamental steps. I am afraid your position only will lead to a collapse and all the effort and good intentions will come to nothing.
I hope you can reconsider your positions again.