Department of Medical Physics,Royal Perth Hospital, Western Australia 6001,
Valendar F. Turner, John Papadimitriou, Barry Page, David Causer, Helman Alfonso
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In his rapid response "For some people it’s April Fool’s day all year round" (3 April 2003) with regards to the Padian study, Peter Flegg wrote "I explicitly stated that "there was a low transmission rate" in her study…". It must be pointed out that in the prospective study of 175 HIV–discordant couples, tested every six months, not one of the 175 non-infected men or women became HIV positive. That is, the prospective Padian study did not prove a low transmission rate but a zero transmission rate, that is, no transmission.
As far as Peter Flegg being perplexed about the thinking of "dissidents", does he expect all scientists to think alike? Science develops by a diversity of views and by questioning existing paradigms. While we agree with the main views held by other "dissidents", that is, there is no proof that HIV cause AIDS, there are also differences. We also agree with some views held by the "HIV" experts including the following:
Peter Flegg also wrote we claim that HIV "spreads through anal intercourse". In fact, in one of our rapid responses we wrote "It is also our view that a positive antibody test ("HIV") is acquired by the same means, that is, passive anal intercourse."( 24th March 2003) In another rapid response we wrote "all the presently available data prove it is only passive anal intercourse which is the risk factor for the acquisition of a positive antibody test." (1st April 2003.) This means that either HIV is the only unidirectionally spread STD in the history of medicine or whatever causes the positive antibody test cannot be an infectious agent.
It is true that in some of our articles, to present our argument we have assumed the existence of HIV in order to show how such an assumption leads to untenable claims for the HIV theory of AIDS.
Peter Flegg wrote that we have been "asserting that HIV does not exist". Peter Flegg may have interpreted our published material in this manner but it is important to state that we have never said HIV does not exist. Rather, from the very beginning when Montagnier and Gallo published their studies in 1983/1984, we have consistently argued that neither their data nor anybody else’s prove that HIV exists. (This issue can be resolved by performing experiments. This is exactly what we argued for and what was accepted by both sides at the Presidential AIDS Panel Meetings held in July 2000, Johannesburg, South Africa http://aidsmyth.addr.com/panelreport.htm). Some of the reasons can be found in what follows.
Following the appearance of AIDS in 1981 many aetiological factors were proposed. In May 1983 Montagnier announced the discovery of a retrovirus, now known as HIV, from lymphatic tissue of a gay man with lymphadenopathy. One year later Gallo reported data which "suggests that HTLV-III [HIV] is the primary cause of AIDS". By 1986 the scientific community accepted the Gallo assertion that the same data was "clearcut evidence" that HIV is the causative agent of the clinical syndrome. Even today the five Science papers published by these French and American groups are still widely regarded as proving beyond all reasonable doubt that HIV exists and is the cause of AIDS.
However, not all scientists accepted these findings. In 1987 Peter Duesberg
published an invited paper in Cancer Research on retroviruses and cancer
in which he also questioned the role of HIV in AIDS. At the same time one of us
(EPE) also challenged the theory including the data claimed to prove the
existence of HIV. Papadopulos-Eleopulos also proposed an alternative,
non-infectious aetiology and treatments based on this hypothesis. Since then our
group has published papers addressing every facet of the HIV theory including a
detailed examination of HIV isolation and the HIV genome. Here we confine
ourselves largely to addressing the data published by Montagnier and Gallo in
their 1983/84 Science papers. Genomic data are only briefly discussed
because the existence of HIV and the HIV theory of AIDS were universally
accepted before such data were available. For a comprehensive discussion on
genomic data the reader is referred to reference 19.
A virus possesses two characteristic properties. The first is anatomical, that is, being a microscopic particle of individual morphology, the second, the ability to generate identical progeny by synthetic processes obligatorily occurring within living cells. It is the latter attribute which defines a particle with the appearances of a virus, that is, a viral-like particle, as infectious and thus a virus. The three subfamilies (0ncovirinae, Lentivirinae and Spumavirinae) of Retroviridae (Retroviruses) are "enveloped viruses with a diameter of 100-120 nm budding at cellular membranes. Cell released virions contain condensed inner bodies (cores) and are studded with projections (spikes, knobs)". The retroviral particles contain RNA and the enzyme reverse transcriptase (RT), an RNA dependent DNA polymerase which catalyses the synthesis of DNA contrary to the central dogma of biology, that is, in a direction "reverse" from DNA to RNA. According to retrovirologists, such DNA is then integrated into existing cellular DNA as a "provirus". Retroviral particles share the property of concentrating (banding) at a density of 1.16 gm/ml when centrifuged at high speeds in sucrose density gradients, a fact long used in their purification.
All retrovirologists agree that to prove the existence of a new retrovirus one must isolate it. However, the term "virus isolation" is beset with semantic difficulties and ambiguities. The dictionary meaning of "isolation" derives from the Latin insulatus (made into an island) and refers to the act of separating an object from all other matter that is not that object. "Purification" means to obtain something free from impurities. In this context isolation is the same as purification. Because virus particles are small it is not possible to obtain a single, isolated particle. The next best thing is to obtain a mass of particles separate from everything else. Until the early 1980s, for the isolation of animal retroviruses as well as the "first" human retrovirus HL23V, by isolation retrovirologists meant purification. On the other hand, nowadays both basic and specialised texts rarely define "isolation" and when they do such attempts are non-illuminating. For example, Levy defines isolation as a "sample of a virus from a defined source", and White as the ability to "identify a totally unforeseen virus, or even discover an entirely new agent". Encompassed as "virus isolation" are listed methods of culturing specimens in tissue and chick embryo cells, as well as live animals, following by documentation therein of cytopathic and pathological effects, haemoabsorption, immunofluorescence, antigen/antibody reactions and "characterisation of the viral genome". HIV experts, including Luc Montagnier and Robin Weiss define "virus isolation" as "propagating them [viruses] in cells in culture" and see www.theperthgroup.com/aids/vftweiss.html. However, if "virus isolation" is to "take a sample of a virus from a defined source", or "propagating them in cells in culture", then first one must have prior proof that a virus exists in "a defined source" or "in cells in culture". One cannot know that a virus exists or define its constituents without purification (isolation) of the putative viral particles.
There are several reasons why this is mandatory:
To prove that the retrovirus-like particles are infectious, that is, they are a virus
The finding of particles with the appearances of retroviruses, is not proof that such particles are retroviruses and even less proof a particle is a particular retrovirus. Particles bearing the morphological characteristics of retroviruses are ubiquitous. In the 1970s such particles were frequently observed in human leukaemia tissues, cultures of embryonic tissues and "in the majority if not all, human placentas". Type-C retroviral particles are present in "fish, snakes, worms, pheasant, quail, partridge, turkey, tree-mouse and agouti" as well as in "tapeworms, insects...and mammals". Gallo was well aware of this problem as far back as 1976 when he wrote: "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". In other words, it is not possible to claim a particle is a retrovirus merely by appearances. To prove that retrovirus-like particles observed in a culture are a virus one must isolate the particles, characterise their proteins and RNA and introduce the particles into a secondary culture. If any particles are released in the secondary culture they too must be isolated and proven that their proteins and RNA are the same as those from the primary culture. In such experiments one must not ignore the use of legitimate controls and in doing so take in consideration an important difference between retroviral and other infectious agents.
When one finds an infectious agent, for example a virus or a bacterium, either in vitro or in vivo, one may be assured that the agent has been introduced into the culture or animal from outside. Retroviruses are the exception. This is because normal human and animal genomes contain information which, under the appropriate conditions, leads to the synthesis of retroviral RNA and proteins, or even to the assembly of retroviral particles, that is, to the expression of endogenous retroviruses. And although as late as 1994 both Gallo and Fauci taught "there are no known human endogenous retroviruses", it is known that at least 1% of the human DNA is retroviral DNA and that endogenous retroviruses are present "in all of us". Furthermore, new endogenous retroviral genomes may arise from rearrangements of existing retroviral genomes, cellular DNA or both, caused by many factors, including pathogenic processes. The expression of endogenous retroviral genomes may arise spontaneously and may be significantly accelerated and the yield increased by conditions which induce cellular activation. According to the eminent retrovirologist George Todaro, "the failure to isolate endogenous viruses from certain species may reflect the limitations of in-vitro cocultivation techniques". Endogenously produced retroviruses are morphologically and biochemically indistinguishable from exogenous retroviruses. Because of this, the finding of identical retrovirus in serially "infected" cultures/cocultures is not proof that the cells are infected with exogenous retrovirus. One method which may assist resolve but will not prove whether cells acquire virus from the outside (exogenously acquired retrovirus, infectious retrovirus) and have not assembled a retrovirus from information already existing in normal cells (endogenous retrovirus), is to conduct control cultures/cocultures in parallel with test cultures/cocultures. The only difference between test and control cultures should be the introduction of tissue assumed infected into the test cultures. In every other respect control cultures must be dealt with identically. For example:
To determine their biological effects
Without recourse to pure particles it is impossible to determine whether effects are due to virus particles or contaminants including "chemical stimulants", a caveat stressed as far back as 1911 by the Peyton Rous, the father of retrovirology.
In 1911 Rous induced malignancy in chickens by injections of cell-free filtrates obtained from a muscle tumour. Similar experiments were repeated by many researchers and the tumour inducing filtrates became known as filterable agents, filterable viruses, Rous agents, Rous virus and ultimately retroviruses. However, Rous himself expressed doubts that the agents which caused the tumours were infectious in nature. Indeed he warned, "The first tendency will be to regard the self-perpetuating agent active in this sarcoma of the fowl as a minute parasitic organism. Analogy with several infectious diseases of man and the lower animals, caused by ultramicroscopic organisms, gives support to this view of the findings, and at present work is being directed to its experimental verification. But an agency of another sort is not out of the question. It is conceivable that a chemical stimulant, elaborated by the neoplastic cells, might cause the tumour in another host and bring about in consequence a further production of the same stimulant".44
To characterise the viral proteins
The only way to prove that a protein is a constituent of an object is to obtain it from that object, or when the object is very small as is the case of viruses, from material consisting of purified virus particles. If the material contains impurities which are proteins or contain proteins, it is not possible to determine which are viral and which are not. Yet only after the viral proteins are characterised is it possible to employ them as antigens in antibody tests.
To characterise the viral genome
As for viral proteins the only way to prove that a stretch of RNA is viral it is to obtain it from material which contains nothing else but virus particles. If the material contains impurities the impurities must not include RNA. Then and only then can the RNA and its complementary DNA (cDNA) be used as probes and primers for genomic hybridisation and PCR studies.
To act as a gold standard for the antibody tests
The reaction of a virus or viral protein with an antibody present in a patient's serum does not prove that the antibody is induced by or directed against the virus or a viral protein. That is, it does not prove the reaction is specific. This is because there are significant obstacles which hinder the interpretation of antibody/antigen reactivity including non-specific stimulation, cross-reactivity or both. Cross reactivity results from antibody molecules, even monoclonal antibodies, interacting not only with the inducing antigen but also with other antigens. Indeed, there are instances where "cross-reactive antibodies may have higher affinity with antigens other than the inducing antigen. Even antigens that differ for most of their structure can share one determinant, and a monoclonal antibody recognising this site would then give a 100% cross-reaction. An example is the reaction of autoantibodies in lupus with both DNA and cardiolipin...It should be emphasised that sharing a "determinant" does not mean that the antigens contain identical chemical structures, but rather that they bear a chemical resemblance that may not be well understood, for example, a distribution of surface charges". Since polyclonal antibodies are composites of monoclonal antibodies these facts apply equally, if not more so, to polyclonal antibodies. These facts have been extensively exploited in clinical medicine for the diagnosis of diseases such as syphilis and infectious mononucleosis. In these diseases, T. pallidum and Epstein-Barr virus cause the appearance of antibodies reactive with ox-heart proteins and sheep and horse red blood cells. However, this does not mean that patients are "infected" with ox-heart, or horse red blood cells and the diseases are induced by these agents. The only way to determine the specificity of an antibody/antigen reaction is to use an independent method, a gold standard to prove the presence or absence of the antigen. The only possible gold standard for a test to prove a virus infection is the virus in question. That is, virus isolation/purification.
METHODS FOR THE ISOLATION/PURIFICATION OF RETROVIRUS-LIKE PARTICLES
Up till the 1950s retroviruses were isolated/purified by filtration although this method was less than satisfactory. With the development of the electron microscope, apparently, for some retrovirologists, the detection of retrovirus-like particles was deemed sufficient to prove the existence of a retrovirus. However, other scientists including the well-known retrovirologist, JW Beard, recognised that cells, including uninfected cells, under various conditions, were responsible for the generation of a heterogenous array of particles some with the appearances of retroviruses. Beard stressed: "identification, characterisation, and analysis are subject to well-known disciplines established by intensive investigations, and the possibilities have by no means been exhausted. Strangely enough, it is in this field that the most frequent shortcomings are seen. These are related at times to evasion of disciplines or to their application to unsuitable materials. As was foreseen, much of the interest in the more tedious aspects of particle isolation and analysis has been diverted by the simpler and undoubtedly informative processes of electron microscopy. While much can be learned quickly with the instrument, it is nevertheless clear that the results obtained with it can never replace, and all too often may obscure, the need for the critical fundamental analyses that are dependent on access to homogenous materials" (italics ours).
By the 1970s there was general agreement that "Virions of RTV [retroviruses] have a characteristic buoyant density, and centrifugation to equilibrium in density gradients is the preferred technique for purification of RTV". The method of banding in density gradients is not ideal either. Substances other than retroviruses may band at the same density. This is why at a meeting held at the Pasteur Institute in 1972, Francoise Barre-Sinoussi and Jean-Claude Chermann stressed that to claim purification of retrovirus-like particles using sucrose density gradients it is absolutely necessary to prove, using the electron microscope, that the 1.16 gm/ml band contains nothing else but particles with "no apparent differences in physical appearances".
THE PHENOMENA CLAIMED TO PROVE THE EXISTENCE OF HIV
In May 1983 Luc Montagnier, Francoise Barre-Sinoussi, Jean-Claude Chermann and colleagues published a paper in Science entitled, "Isolation of a T-Lymphotropic retrovirus from a patient at risk for Acquired Immune Deficiency Syndrome (AIDS)". This is the paper which, since the resolution of the polemics between Montagnier and Gallo concerning allegations of misappropriation by the latter of the French virus sent to the US by the Pasteur Institute, is accepted as being the study which proved the existence of HIV. There it was shown that mitogen stimulated lymph node cell cultures from a gay man (BRU) with lymphadenopathy were able to transcribe the synthetic RNA primer-template An.dT15. From this data Montagnier and his colleagues concluded that BRU’s lymph node cells were infected with a retrovirus. The finding of the same activity in the supernatant of a coculture consisting of the same cells with stimulated lymphocytes from a healthy individual was considered proof for virus transmission as well as isolation. In another experiment supernatants from the cocultures were added to two, three day old, stimulated umbilical cord lymphocytes cultures. "Electron microscopy of the infected umbilical cord lymphocytes showed characteristic immature particles with dense crescent (C-type) budding at the plasma membrane". Supernatant from the culture was banded in sucrose density gradient and the 1.16 gm/ml band was shown to transcribe An.dT15. The proteins in the 1.16 gm/ml band as well as the proteins of a cellular extract were separated according to their molecular weight using "denaturing buffer and electrophoresed on 12.5 percent polyacrylamide-SDS slab gel". When the strips were incubated with human sera many proteins from the cellular extracts were found to react with serum from BRU, another gay man as well as a "healthy donor". In the strips containing the proteins from the 1.16 gm/ml band three proteins including a p25 (‘p’ for protein, 25 for its molecular weight in thousands) were found to react. They also reported that the p25 did not react with antibodies to HTLV-I. The material banding at 1.16 gm/ml was claimed to be "purified, labelled, virus" although no electron microscopic data were presented. The authors concluded: "A retrovirus belonging to the family of recently discovered human T-cell leukaemia viruses (HTLV), but clearly distinct from each previous isolate, has been isolated from a Caucasian patient with signs and symptoms that often precede the acquired immune deficiency syndrome (AIDS). This virus is a typical type-C RNA tumor virus, buds from the cell membrane, prefers magnesium for reverse transcriptase activity, and has an internal antigen (p25) similar to HTLV p24".
Robert Gallo and his associates did not consider the Montagnier group data as proving "true isolation". As late as 1997, in a book published by one of the best known HIV experts, Jaap Goudsmit, one reads: "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 enzyme is not even specific to retroviruses much less to Lentiviruses (see below)]...The BRU virus grew slowly and with difficulty, but its identity and activity were reported in the May 20, 1983 issue of Science...The Pasteur Group was widely acclaimed but very worried. In the world of virology, finding a new virus is not enough: You must propagate and isolate the organism for analysis by other virologists. The French had not yet isolated their new lentivirus".
Why then did: (a) Gallo, who reviewed the Montagnier manuscript, recommend its publication? (b) all the HIV experts including Gallo and Goudsmit (on page 24 of his book one reads: "BRU was the first strain to be isolated") accept that the first isolation of HIV and thus of its existence was proven in the May 1983 Science paper?
A year later, in May 1984, Gallo, Popovic and their colleagues published four papers in Science in which they claimed "isolation" of another retrovirus from AIDS patients. However, in addition to the use of a leukaemic cell line, the only difference between the Montagnier and Gallo groups’ data were quantitative. In the first paper entitled "Detection, Isolation and Continuous Production of Cytopathic Retroviruses (HTLV-III) from Patients with AIDS and pre-AIDS", (HTLV-III=HIV), experiments were described in which "concentrated culture fluids harvested from short-term [mitogenically stimulated] cultures of T-cells" from patients with AIDS or pre-AIDS were cultured with a mitogenically stimulated leukaemic cell line HT and highly selected clones obtained by culturing HT with irradiated cells of a healthy donor. The data presented as proof of isolation of HIV were: (a) RT activity in cell free supernatants and the 1.16 gm/ml band; (b) reaction in the cultures with "Rabbit antiserum to HTLV-III" and "Patient serum (E.T.)"; (c) EM showing the presence of retroviral-like particles in the cultures.
An enquiry conducted by the National Institutes of Health Office of Scientific Integrity found that the HT cell line was cultured not with concentrated fluids (supernatant) originating from individual AIDS patients, but with concentrated fluids pooled initially from individual cultures of three patients and ultimately from the individual cultures of ten patients. In evidence given to this enquiry the reason given was because none of the supernatants "individually was producing high concentrations of reverse transcriptase". In other words, Gallo and his colleagues did not regard the levels of RT from individual cultures as proof that individual specimens contained a retrovirus. The Gallo investigation found the pooling of specimens to be "of dubious scientific rigor". One scientist described the procedure as "really crazy". Most importantly how could Gallo use the reaction with rabbit antiserum to prove HIV isolation (purification) when, to obtain rabbit antiserum rabbits must first be injected with pure HIV? It is inexplicable how Gallo and his colleagues could already possess "Rabbit serum to HTLV-III" before they had proved the existence of a novel virus. However, if the antiserum was manufactured "from rabbits infected repeatedly with disrupted HTLV-III", that is, the material banding at 1.16 gm/ml, one would expect to obtain antibodies to all the proteins constituting this material even if the proteins were cellular and not viral.
In the second paper entitled "Frequent Detection and Isolation of Cytopathic Retroviruses (HTLV-III) from patients with AIDS and at risk of AIDS", Gallo and his colleagues claimed to have "isolated" HTLV-III (HIV) from 26/72 (36%) of AIDS patients. In this paper isolation was defined as "more than one of the following": "repeated detection of a Mg2+-dependent reverse transcriptase activity in supernatant fluids; virus observed by electron microscopy (EM) [retrovirus-like particles in the culture]; intracellular expression of virus-related antigens detected with antibodies from seropositive donors or with rabbit antiserum to HTLV-III; or transmission of particles". Transmission of particles was defined as "detected by RT assays or by electron microscopic observation, to fresh human [umbilical] cord blood, bone marrow, or peripheral blood T lymphocytes", cultured with supernatants from the "infected" cultures. (It can be seen that the Gallo group method permitted "isolation" of a retrovirus without evidence for either particles or RT).
In the third paper, proteins from the 1.16 gm/ml band which they claimed was "purified" HIV, as well as the proteins from the "infected" cells, were "lysed and fractionated by electrophoresis on a 12% polyacrylamide slab gel in the presence of SDS. The protein bands were electrophoretically transferred to a nitrocellulose sheet" and reacted with different sera. In other words Gallo used a technique which is known as Western blot (WB) antibody test. Many proteins from the cellular extract were found to react with sera from both patients and healthy individuals. They also reported that two proteins from the 1.16 gm/ml band, p24 and p41, reacted with patient sera. For this and no other reason it was claimed that "these molecules are the major components of the virus preparation. p24 and p41 may therefore be considered the viral structural proteins". As far as morphology is concerned, the Gallo group reported that the HIV particle "is produced in high numbers from infected cells by budding from the plasma membrane. A possible unique feature of this virus is the cylindrical shaped core observed in many mature virions...HTLV-III is a true member of the HTLV family". (HTLVs are type C retroviral particles and not Lentiviruses as HIV is claimed to be).
In the fourth paper, instead of separating the proteins which banded at 1.16 gm/ml and then incubating them with patient sera, the mixture of all proteins was used, that is, they performed a test known as enzyme-linked immunosorbent assay (ELISA). "To understand the molecular nature of the antigens recognised by ELISA", they also performed WB, with some of the sera. They reported that: "Serum samples from 88 percent of patients with AIDS and from 79 percent of homosexual men with signs and symptoms that frequently precede AIDS, but from less than 1 percent of heterosexual subjects, have antibodies reactive against antigens of HTLV-III. The major immune reactivity appears to be directed against p41, the presumed envelope antigen of the virus…The data presented here and in the accompanying reports suggest that HTLV-III is the primary cause of AIDS". Two years later Gallo wrote that "The results presented in our four papers provided clearcut evidence that the aetiology of AIDS and ARC was the new lymphotropic retrovirus, HTLV-III"67 (italics ours).
As mentioned, apart from the quantitative difference, the Gallo group experiments are no different from those performed by Montagnier and his colleagues. It follows then, that if Montagnier's group data do not prove "true isolation" neither did Gallo's nor anybody else’s because to date everybody has repeated (for the vast majority only part thereof) the same experiments as these two groups. It is of pivotal significance that neither group reported the use of valid controls (see above) nor did they prove they had obtained purified retrovirus-like particles. The question then is, do the data obtained by the Montagnier and Gallo group, that is RT, particles in culture and antigen/antibody reactions prove the isolation of a unique, or even a retrovirus, any retrovirus?
Without doubt, if by isolation one expects proof of purification, then the detection of an enzyme, or retrovirus-like particles in a culture, or proteins either in the cells or the 1.16 gm/ml band which react with antibodies present in human or animal sera, do not comply. To argue otherwise one must define the detection of cardiac or hepatic enzymes in the blood of patients suffering chest pain or jaundice as proof for isolation of the human heart or liver. Likewise, if an antibody/antigen reaction is proof for isolation of a virus then antibody reactivity to the protein βHCG is proof for isolation of the human placenta. The assertion that the detection of a retrovirus-like particle in a culture proves isolation is no different from asserting that the detection of a fish-like creature in the ocean is the same as having a definite fish in your frying pan. The detection of RT, retrovirus-like particles and antibody/antigen reactivity can only be considered proof for the detection of a retrovirus, and then if, and only if, there is prior knowledge that the three phenomena are specific to retroviruses.
To claim that the detection of this phenomena proves the existence of a new retrovirus one must have proof that at least one of the three phenomena is different from that observed in all other known retroviruses.
At present some of the leading HIV researchers consider RT as being the "sine qua non" of retroviruses and regard the detection of reverse transcription in lymphocyte cultures from AIDS patients not only as proof of the presence of such viruses but of HIV itself including HIV isolation and quantification. However, according to some of the best known retrovirologists including its discoverer, as well as the Nobel Laureate and former Director of the US National Institute of Health Harold Varmus, reverse transcriptases are present in all cells as well as bacteria and viruses. "Reverse transcriptase (RT) was first discovered as an essential catalyst in the biological cycle of retroviruses. However, in the past years, evidence has accumulated showing that RTs are involved in a surprisingly large number of RNA-mediated transcriptional events that include both viral and nonviral genetic entities". Even if RT were a property only of viruses it is not specific to retroviruses. According to Varmus, "Reverse transcription was assigned a central role in the replication of other viruses [hepatitis B and cauliflower mosaic viruses] and in the transposition and generation of other kinds of eukaryotic DNA". "The hepatitis B viruses (HBVs) are small DNA viruses that produce persistent hepatic infections in a variety of animal hosts and replicate their DNA genomes via reverse transcription of an RNA intermediate. All members of this family contain an open reading frame (ORF), "P" (for pol), which is homologous to retroviral pol genes" [pol=polymerase]. "Hepatitis B virus (HBV) resembles retroviruses, including HIV, in several respects. In particular, both viruses contain reverse transcriptase, and replicate through an RNA intermediate". Because of this, it has been suggested that hepatitis B infection should be treated with the same antiretroviral agents as HIV infection.
At present, evidence exists which shows that although the major target organ for hepatitis B virus is the liver, cells other than hepatocytes "including peripheral blood lymphocytes and monocytes, may become infected with HBV". Lymphocyte stimulation in general and PHA (an agent employed in the majority of cultures with tissues from AIDS patients), is associated with the production of hepatitis B virus from peripheral blood lymphocytes in patients infected with HBV including "viral replication in chronic hepatitis B infection of childhood". Hepatitis B virus infection is widespread in the AIDS risk groups.
In the early 1970s Gallo proved that cultures of leukaemic cells transcribe the An.dT15 template-primer as does material banding at 1.16 gm/ml originating from "PHA stimulated (but not unstimulated normal human blood lymphocytes". Reading the Gallo 1984 Science papers the impression is gained that the leukaemic HT cell line and thus its clones, including H9 which Gallo used, was a new cell line developed in Gallo's laboratory. However it is now known that the HT cell line is the HUT78 cell line which originated from a patient with adult T cell leukaemia, a disease which Gallo claims is caused by the retrovirus, HTLV-I. A year earlier Gallo claimed to have proven that the HUT78 cells are infected with HTLV-I. If this is the case, the Gallo group would have detected RT in their cultures even if the enzyme is specific to retroviruses and the cultures were not infected with HIV. Other researchers reported RT activity in normal non-infected spermatozoa.
It must also be pointed out that the presence of HIV reverse transcriptase was detected by both Montagnier and Gallo, (and all HIV experts since) indirectly, that is, by detecting the transcription of the template-primer An.dT15. However, at least in 1984 if not 1983, Montagnier and his colleagues knew that in the 1970s there was proof that "Among a number of template primers, (rA)n.(dT)12-18 has been most frequently employed since RT shows high activity with this template primer. However, the problem is that the cellular DNA polymerases (pol b and pol g ) also effectively utilize the same template primer". In fact, in 1975, an International Conference on eukaryotic DNA polymerases defined DNA polymerase γ as the cellular enzyme which "copies An.dT15 with high efficiency but does not copy DNA well". One year earlier Gallo wrote: "Under appropriate reaction conditions DNA polymerase β can efficiently transcribe poly (A) primed by oligo (dT)". Thus it is possible to detect transcription of An.dT15 even when no RT, either viral or cellular, is present, especially under the conditions used to prove the existence of HIV. (Both Montagnier and Gallo accept that the phenomena detected in cultures which are said to prove HIV isolation cannot be detected unless the cells are chemically stimulated). Nowadays the non-specificity of RT is broadcast even in the popular press to readers contemplating the purchase of shares in biotechnology companies.
In conclusion, even if one accepts Montagnier and Gallo's groups’ definition of isolation, given that RTs and reverse transcription are nonspecific to retroviruses, their detection even in an unlimited number of consecutive cultures/co-cultures cannot be considered proof for isolation and propagation of a retrovirus.
Montagnier and his colleagues reported HIV initially as a type C particle, then as a type D particle and then as a Lentivirus. In 1984 Gallo and his colleagues reported HIV as a type C particle. However, in 1985 Gallo wrote: "A possible unique feature of the virions is the cylindrical core observed in many presumably mature virions. Virions having this type of core have been frequently reported for certain type D retroviruses, and in some instances, for type C retroviruses". Jay Levy reported HIV as a type D particle. Others at the University of California wrote that "AIDS virus isolated show morphologic characteristics of type C, type D and Lentiviruses". According to Anthony Fauci and others" "T-cells and macrophages handle the virus very differently. In the T-cell, virus buds out of the external plasma membrane of the cell. In the monocyte/macrophage cultures it buds into membrane-bound vesicles inside the cells". The latter is a description of a type A, retroviral particle. Thus the leading HIV experts have described HIV as a member of two subfamilies and three genera of Retroviridae. These taxonomical differences imply that if HIV was a newly discovered mammal, it could have been either human, a gorilla or an orang-utan. By consensus at present HIV is regarded as a Lentivirus. This agreement was reached when it was realised that in "HIV" positive individuals AIDS did not appear soon after "infection", although the designation Lentivirus is a morphological description.
"HIV infected cultures" contain in addition to the particles with the morphology attributed to HIV many other "viral particles". For example: Hockley and his colleagues from the Electron Microscopy and Photography Section and Division of Virology at the National Institute for Biological Standards and Control in the United Kingdom describe a profusion of particles which they divide broadly into three groups, mature, ring-like and small with spikes. The mature particles "were approximately spherical in shape and 100 to 150nm in diameter. The outer lipid membrane was frequently broken or absent in places, and there was no evidence of surface spikes...A few mature particles were found that were larger than average and appeared to contain a double nucleoid...in the preparation of HIV there were always many vesicles with granular contents in which it was not possible to recognise a distinct nucleoid". Also, "The ring-like particles had a more consistently spherical shape and were larger (140nm in diameter)" and the small particles "were unusually spherical but sometimes slightly angular in shape and 65 to 90nm in diameter" and had spike-like projections on their surface. Hans Gelderblom who has done most of the EM studies in HIV/AIDS research reported that although Lentiviruses and thus HIV is considered to have a cone shaped core, he and his colleagues found centrosymmetric and tubular cores as well. The caption to one photograph reads: "Virions can be seen having either elongated, 'baton-like' tubular cores 30-35nm in diameter or containing more than one core. Tubular and regular cone-shaped cores can coexist within one virion". The text states: "Rarely, tubular core structures reminiscent of batons with a diameter of 30-35nm and a length of 150-250nm are observed". Lekatsas and other virologists from Pretoria and Johannesburg reported: "We used the characteristic cylindrical structure in the core as an identifying characteristic for the virus to distinguish it from cellular debris and also noted that it may vary considerably in its dimensions and morphological features. We have found two basic virus particle sizes, 90nm and 120nm, both present in large numbers. The larger particle bears no surface projections while the smaller particle is rarely 'naked' and usually bears projections". The US CDC reported: HIV particles are "usually round and have a diameter of about 85-95nm...Virus with bar-shaped nucleoids and particles with a tear-drop shape are commonly seen in HTLV-III/LAV infected lymphocytes, sometimes ring-shaped particles without dense nucleoids are also seen". The question then arises if the particles with the "unique" morphology considered to be HIV represent an exogenous retrovirus originating from tissues of AIDS patients or those at risk, then what is the origin and role of the many non-HIV particles and which, if any, of the "HIV" or non-HIV particles band at 1.16 gm/ml? That is, which have the density characteristic of retroviruses? Retrovirus-like particles have been found in non-HIV-infected cord blood lymphocytes cell cultures and in cells used for HIV "isolation" such as and H9 (HUT-78), CEM, C8166 and EBV transformed B-cells. Retrovirus-like particles antigenically related to HIV have been found in cultures of salivary gland extracts from patients with Sjorgen's syndrome. In the only EM study, either in vivo or in vitro in which suitable controls were used and in which extensive blind examination of controls and test material was performed, particles indistinguishable from "HIV" were found in 18/20 (90%) of AIDS as well as in 13/15 (87%) of non-AIDS related lymph node enlargements. This led the authors to conclude: "The presence of such particles do not, by themselves indicate infection with HIV".
It is of pivotal significance to note that:
Hans Gelderblom and his colleagues have estimated that immediately after being released from the cell membrane "HIV particles" possess an average of 0.5 knobs per particle which are rapidly lost, but also pointed out that "it was possible that structures resembling knobs might be observed even when there was no gp120 [knobs] present, i.e. false positives". Yet all HIV experts agree that the infectivity of the HIV particles is determined by the gp120 (knobs). Thus, according to Montagnier et al, "The gp120 is responsible for binding the CD4 receptor" while for Matthews and Bolognesi, "First gp120 binds to the CD4 receptor on an uninfected cell; then gp41 becomes anchored in the adjoining membrane; next the two membranes begin to fuse, and the virus spills its contents into the cell". Callebaut et al state, "The human immunodeficiency virus (HIV) infects lymphocytes, monocytes, and macrophages by binding to its principal receptor, the CD4 molecule, through the viral envelope glycoprotein gp120. The V3 loop of gp120 is critical for HIV infection". Others are in complete agreement.
The general agreement that gp120 (knobs) is absolutely necessary for HIV particles to be infectious and the fact that this protein (knobs) is not present in the cell free particles leads to one unavoidable conclusion, that is, the "HIV" particles are not infectious, they are not viral particles. Furthermore,
The only conclusion one can draw from the electron microscopy data is that the reported particles are non-HIV or even retroviral specific which means that the detection of such particles, be it in an unlimited number of consecutive cultures/cocultures, is not proof for isolation, no matter how isolation is defined.
It has already been mentioned that the mere interaction between an antigen such as a protein or a virus and an antibody does not prove any more than a chemical relationship. In all other aspects the reaction may be totally nonspecific. To prove the specificity of the reaction one must employ an alternative, independent method of proving the existence of the antigen, that is, one must use what is generally known as a gold standard. The only possible gold standard for the HIV antibody tests (the antigen/antibody reaction) is HIV itself, that is, HIV isolation. No matter how one defines isolation, obviously the definition cannot include the antibody/antigen reaction (the antibody test, neither the WB nor the ELISA) because, by doing so, not only does one not have an independent analysis, the test becomes its own gold standard and is thus rendered meaningless. Reading the 1984 Gallo Science papers it appears that Gallo always defined isolation as "more than one of the following: reverse transcriptase activity either in the supernatant or the 1.16 gm/ml band; retrovirus-like particles in the culture or reaction between proteins (either in the cultured cells or the 1.16 gm/ml band) with antibodies present in the patients sera or "antiserum to HTLV-III". However, in an interview Gallo gave to Huw Christie, the editor of the British magazine Continuum, at the 1998 Geneva AIDS Conference, Gallo said, "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. That's all I can tell you. It was an experimental tool when we added it and for us it worked well because we could isolate the virus when we did it". In other words:
In an interview which Montagnier gave to the French Journalist Djamel Tahi in 1997, he stated: "analysis of the proteins of the virus demands mass production and purification. It is necessary to do that". Indeed, the only way to prove that a protein is a viral constituent is to obtain it from material which contains nothing else but viral particles. Instead Montagnier’s and Gallo's groups incubated the proteins which banded at 1.16 gm/ml with sera from AIDS patients. The proteins which were found to react more often with the sera were said to be HIV proteins (although neither group published any evidence for the existence of retrovirus-like particles, pure or impure, at the 1.16 gm/ml band); and the antibodies to be HIV antibodies. However, even if the antigen/antibody reaction is 100% specific, that is, antibodies react only with inducing antigens and with no other, from such a reaction it is impossible to determine the origin even of one reactant much less the origin of both the antigen and the antibody. Let us consider differing scenarios:
We know that it is not possible to take a protein from the 1.16 gm/ml mass and know from which component it originates. Let us then follow the same steps as Gallo and Montagnier. Take the mass of material banding at the 1.16 gm/ml, disrupt the proteins and electrophoretically position the proteins in a strip according to their molecular weights. Using this technique we are unable to state from which component of the 1.16 gm/ml band mass a protein on the strip derives. Next we incubate the proteins in the strip with patient sera and discover that some of the proteins react with antibodies present in the sera. Even if the antibody/antigen interaction is 100% specific such a reaction does permit us to define the origin of the proteins. Yet from such a reaction and without any proof that the antibodies specifically reacted, Montagnier and Gallo defined the origin of both the proteins and the antibodies as "HIV".
As mentioned, Montagnier found three proteins in the 1.16 gm/ml band which reacted with antibodies present in his patients’ sera. These were p25, p80 and p45. Montagnier concluded that his patients were infected with a retrovirus which "contains a major p25 protein, similar in size to that of HTLV-I", but made no comment in regard to the p80 protein. Regarding p45 he wrote: "The 45K [p45] protein may be due to contamination of the virus by cellular actin".
In 1997 Montagnier said that the protein he detected in 1983 had a molecular weight of 43,000 and was actin. (The molecular weight of actin is neither 45,000 nor 43,000 but 41,000. However since Montagnier and Gallo determined the molecular weights of the proteins by their migration in an electrophoretic strip, and because the migration may be influenced by other factors, for example, by the protein's charge, it is possible that these slight differences in the molecular weight are simply the result of experimental variation).
When Gallo and his colleagues used the cellular proteins as antigens in the WB they reported: "The most prominent reactions were the antigens of the following molecular weights: 65,000, 60,000, 55,000, 41,000 and 24,000. Antigens with molecular weights of approximately 88,000, 80,000, 39,000, 32,000, 28,000 and 21,000 gave less prominent reactions…A large protein with a molecular weight of approximately 130,000 and a protein of 48,000 were also detected". In another experiment the "antigens from virus purified from the culture fluids", that is, the 1.16 gm/ml band, were incubated with different sera. They found an "Extensive" reaction of the AIDS patients sera with p24 and p41 and concluded: "these molecules are the major component of the virus-preparation. P24 and p41 may therefore be considered viral structural proteins…Furthermore, an antigen with a molecular weight of approximately 110,000 was detected in the virus preparation but was below limit of detection in the cells. Also, p39 was present in the virus preparation…Occasionally an additional set of antigens was recognised by a serum but their relation to the antigens described above is unclear".
Between 1983-87 the detection of antibodies in patient sera which reacted with p24 or p41 (Montagnier considered reaction with p24 and Gallo with p41 HIV specific) was considered proof that the patient was infected with HIV. In the same period of time it became obvious that a significant number of individuals at no risk of AIDS had antibodies which reacted with these proteins. Since 1987 most of the proteins which Gallo found to react either in the cell extracts or the 1.16 gm/ml band are now considered HIV proteins and laboratories require the presence of antibodies which react with more than one protein before the patient is considered infected with HIV. The number and identity of antibody/protein (Western blot) bands required vary from continent to continent, from country to country and even between and within laboratories in the same country. Thus it is possible for the same patient to be HIV seropositive in New York for example, but not in Africa or Australia.
Furthermore, for many years evidence exists which shows that p24 is not "HIV" specific123,124, p41 is the cellular protein actin,58,125,126 p32 is class II histocompatability DR protein126,127 and p120 / p160 are oligomers of p41.128
Even if proof exists that the "HIV" proteins do indeed belong to a unique exogenous retrovirus, it cannot be assumed that antibodies that react with them are diagnostic of "HIV" infection. To prove the specificity of the antibody tests it is mandatory to:
To date nobody has published such studies using any definition of "HIV" isolation.
The inescapable conclusion is that the antibody/antigen reaction is not HIV specific and thus cannot be used to prove HIV isolation, no matter how isolation is defined.
MORE RECENT DEVELOPMENTS
By 1997 some of the best known HIV experts accepted that:
In that year, in Virology, two studies were published, one by a USA team, principal author Julian Bess, and the other by a Franco-German group, principal author Pablo Gluschankof, with the first electron micrographs of "purified HIV". While in the Gluschankof et al studies the EMs were from the 1.16 gm/ml band, in the Bess et al band they were from pooled bands. The authors of both studies claimed that their "purified" material contained some particles with the appearances of retroviruses and in fact were HIV particles. But they admitted that their material predominantly contained particles which were not retroviruses but "budding membrane particles frequently called microvesicles" or "mock virus". Indeed the caption to the Gluschankof et al electron micrograph reads, "Purified vesicles from infected H9 cells (a) and activated PBMC (b) supernatants", not purified HIV. In further experiments the supernatant from non-infected cultures was also banded in sucrose gradients. Both groups claimed that the banded material from these cultures contained only microvesicles, "mock virus" particles, but no HIV. Both the "HIV" particles and the mock-virus particles possessed membranes. In the USA study the "HIV-1 particles" were differentiated from the microvesicles "by the electron dense cores", whereas in the other study the "HIV" particles were "identified by the relatively homogenous diameter of about 110 nm, the dense cone-shaped core, and the "lateral bodies". However, in the arrowed particles which are said to be HIV it is difficult if not impossible to locate any which have cone-shaped cores or bilateral, "lateral bodies". In fact no particle in any study has the principle morphological characteristic of retrovirus, a diameter of 100-120 nM and surface spikes, knobs. In the Franco-German study the average "HIV" particle diameter is 136 nM and no particle had a diameter less than 120 nM. In the USA study the corresponding dimensions are 236 nM and 160 nM. In other words, the American "HIV" is twice the diameter of the European "HIV", and all other "HIV" particles. The US authors did not note or explain this discrepancy and "were much more focused on showing the mixture of particles in the preparations as opposed to their actual diameters". The diameter of the microvesicles "range in size from about 50 to 500 nm". Both the "HIV" and the "mock" virus particles contained RNA. The RNA of the latter had contained mRNA which is known to be rich in adenine and which, according to Gallo is specific to retroviral RNA. According to Gluschankof et al, "The vesicles contain large amounts of protein and nucleic acids which are unstructured and thus are transparent by electron microscopy", that is why many, but not all, appear "empty" by electron microscopy, while the nucleic acids in the "HIV" particles are structured and for this reason they appear to have an "electron dense cores". However, according to a leading retrovirologist, John Bader, the core density can be changed by external conditions, that is, the culture conditions. It is well known that a structural virus particle or virus-like particle can become "unstructured" in the presence of reducing agents. The possibility cannot be excluded then, that the apparent morphological differences between the two types of particles may be due to nothing more than the difference in the redox of the microenvironment in which they are assembled, released or both. It is significant that actin polymerisation (or actin/myosin interaction) "mediates HIV budding" and release. Evidence also exists that:
The minimum absolutely necessary but not sufficient condition to claim that what are called "HIV-1 particles" are a retrovirus and not cellular microvesicles is to show that the sucrose density fractions obtained from the "infected" cells contain proteins which are not present in the same fractions obtained from non-infected cells. However, Bess et al have shown this is not the case. The only difference one can see in their SDS-polyacrylamide gel electrophoresis strips of "purified virus" and "mock virus" is quantitative, not qualitative. This quantitative difference may be due to many reasons including the fact that there were significant differences in the history and the mode of preparation of the non-infected and "infected" H9 cell cultures, in addition to the "infection".
A similar finding was reported by the same authors a few years earlier. However, while in both studies the proteins of molecular weight "near 42 kDa" (42,000) are labelled as "Actin" and "in the 30- to 40-kDa range" as "HLA DR", all the proteins with molecular weight higher than approximately 42,000 and lower than approximately 30,000 are left unlabelled in the earlier paper. In the 1997 study, three proteins of molecular weight lower than 30,000 are labelled as p24CA, p17MA, and p6/p7NC and are said to represent "major bands of viral proteins". However, also according to the authors, "these labels were added when one of the reviewers asked for them. He felt it would help orient readers when looking at the figure - the reviewer is correct. We did not determine the identities of the bands in this particular gel".
In their earlier study the researchers from the USA presented further evidence that the "viral proteins" were nothing more than cellular proteins. In their efforts to make an HIV vaccine they immunised macaques with, amongst other antigens, "mock virus", that is, sucrose density banded material from the supernatants of non-infected H9 cell cultures. After the initial immunisation the monkeys were given boosters at 4, 8 and 12 weeks. The animals were then challenged with "SIV" propagated either in H9 cells or macaque cells. When the WBs obtained after immunisation but before "SIV" challenge were compared with the WBs post-challenge, it was found that challenge with "SIV" propagated in macaque cells had some additional bands. However, the WBs obtained after the challenge with SIV propagated in H9 cells were identical with the WBs obtained after immunisation but before challenge. In other words, the protein immunogens in the "virus" were identical with the immunogens in the "mock virus". Since both the "mock virus" and "purified" virus contain the same proteins, then all the particles seen in the banded materials including what the authors of the 1997 Virology papers call "HIV" particles must be cellular vesicles. Since there is no proof that the banded, "purified virus", material contains retrovirus proteins and thus retrovirus particles then there can be no proof that any of the banded RNA is retroviral RNA. When such RNA (or its cDNA) is used as probes and primers for hybridisation and PCR studies, no matter what results are obtained, they cannot be considered proof for infection with a retrovirus, any retrovirus.
In the interview which Montagnier gave to Djamel Tahi he was asked why they did not publish an electron micrograph of the 1.16 gm/ml band to prove that the band represented "purified" virus, as they claimed. He replied that the reason for this was that even after "Roman effort" in their "purified" virus they could not see any particles with the "morphology typical of retroviruses. They were very different. Relatively different". When Montagnier was asked if Gallo isolated HIV he replied: "I do not believe so". If there were no retrovirus-like particles in Montagnier’s "purified" virus, then obviously Montagnier and his colleagues could not claim to have isolated a specific exogenous retrovirus, HIV.
THE "HIV GENOME"
To claim that the stretch of RNA (cDNA) is the genome of a unique retroviral particle, HIV, the most basic requirement is proof for the existence of a unique molecular entity "HIV RNA" ("HIV DNA") that is, a unique fragment of RNA (DNA) identical in both composition and length in all infected individuals. The claim that a stretch of RNA (cDNA) is a unique molecular entity which constitutes the genome of a unique retrovirus can be accepted if and only if it is shown that the RNA belongs to particles with the morphological, physical and replicative characteristic of retroviral particles. Proof of this can only be obtained isolating the particles, that is, by obtaining them separate from everything else (purifying them). In 1984 both Gallo’s and Montagnier’s groups reported finding polyadenylated (adenine rich) RNA (poly(A)-RNA) in the 1.16 gm/ml band material obtained from "infected" cultures. The RNA was claimed to be HIV RNA that is, the HIV genome, and its complementary DNA, the HIV provirus. However,
Furthermore, despite the ample evidence to the contrary, for most retrovirologists the finding of a novel nucleic acid in a cell can be due to nothing else but an infectious agent. Half a century has passed since the Nobel Laureate Barbara McClintock discovered the phenomenon of transposition which can lead to the appearance of new genotypes and phenotypes. According to McClintock, the genome can be restructured not only by transposition but also by other means as well. In her Nobel lecture of 1983, she said, "rapid reorganisation of genomes may underline some species formation. Our present knowledge would suggest that these reorganizations originate from some "shock" that forced the genome to restructure itself in order to overcome a threat to its survival...Major genomic restructuring most certainly accompanied formation of new species". The "genomic shock" which leads to the origin of new species may be "either produced by accidents occurring within the cell itself, or imposed from without such as virus infections, species crosses, poisons of various sorts, or even altered surroundings such as those imposed by tissue culture. We are aware of some of the mishaps affecting DNA and also of their repair mechanisms, but many others could be difficult to recognize. Homeostatic adjustments to various accidents would be required if these accidents occur frequently. Many such mishaps and their adjustments would not be detected unless some event or observation directed attention to them...Unquestionably, we will emerge from this revolutionary period with modified views of components of cells and how they operate, but only however, to await the emergence of the next revolutionary phase that again will bring startling changes in concepts".
As we have mentioned elsewhere9,19 an exogenous retrovirus is only one possible explanation for the finding of novel nucleic acids in AIDS patients. Other explanations are:
A basic principle of molecular biology is that the primary sequence of RNA faithfully reflects the primary sequence of the DNA from which it is transcribed. However, in the 1980s RNA editing, "broadly defined as a process that changes the nucleotide sequences of an RNA molecule from that of the DNA template encoding it", was discovered. In the process a non-functional transcript can be re-tailored, producing a translatable mRNA, or modify an already functioning mRNA so that it generates a protein of altered amino acid sequences. Sometimes editing is so extensive that the majority of sequences in a mRNA are not genomically encoded but are generated post-transcriptionally producing the "paradoxical situation of a transcript that lacks sufficient complementarity to hybridize to its own gene!". According to Nancy Maizels and Alan Weiner from the Department of Molecular Biophysics and Biochemistry at Yale University, "the central dogma has survived hard times. The discovery of reverse transcriptase amended but did not violate the central dogma of how genes make proteins; introns qualified the conclusion that genes are necessarily collinear with the proteins they encode; somatic rearrangement of lymphocyte DNA called stability of eukaryotic genomes into doubt...and catalytic RNA challenged the pre-eminence of proteins and breathed new life into the ancient RNA world". However, the discovery of RNA editing "could come close to dealing it a mortal blow".
Thus the finding of novel RNAs in human cells, especially those of AIDS patients and those at risk, can no longer be regarded as incontrovertible proof that the RNA has been exogenously introduced by a putative HIV or any other infectious agents. That this may be the case has of late been accepted by Luc Montagnier. In a written testimony dated February 2nd 2000, to the US House of Representatives Committee on Government Reform, Subcommittee on National Security, Veterans Affairs and International Relations, in support of the work of his colleague, Howard B Urnovitz, (Montagnier is on the scientific advisory board of a publically traded biomedical company whose director is Urnovitz), Montagnier wrote: "I have reviewed Dr Urnovitz's published research and the testimony prepared for presentation to this Committee and strongly advise that future research on Gulf War Syndrome should include the study of the detected genetic material".
Urnovitz and his colleagues presented evidence of the existence, in Persian Gulf War veterans, of "novel", "nonviral" RNAs, "possibly induced by exposure to environmental genotoxins". They concluded: "The patterns of the occurrence of RPAs [polyribonucleotides] in the sera of GWVs [Gulf War Veterans] and healthy controls are sufficiently distinct to suggest possible future diagnostic applications…Our studies of patients with active multiple myeloma suggest that patients with individual chronic multifactorial diseases may have unique RPAs in their sera. Validated tests for such putative surrogate markers may aid in the diagnosis of such diseases or in the evaluation of responses to therapeutic modalities".
In the same year that Montagnier claimed HIV isolation, 1983, in a paper entitled "Expression of novel transposon-containing mRNAs in human T cells" researchers from the USA and Canada reported "Using an HERV-H LTR probe, 6 and 4.5 kb transcripts were detected by Northern blot analysis which were induced in normal peripheral T cells after treatment with phytohaemagglutin" (HERVº human endogenous retrovirus). Phytohaemagglutin has been and continues to be used not only by Montagnier but by virtually every retrovirologist who claims proof for HIV isolation.
Since Montagnier agrees with Urnovitz that novel, nonviral RNAs appear in the Gulf War Veterans, then why should the existence of novel RNAs:
When hard pressed all the HIV experts will ultimately accept the non-specificity of retroviral-like particles, reverse transcription and antibody/antigen reactions. That this is the case is illustrated is exemplified by the fact that at the beginning of the AIDS era they were the first to report isolation of HIV from individuals at no risk of AIDS.166-168 In 1985 Weiss and his colleagues reported the isolation of a retrovirus from mitogenically stimulated T-cell cultures of two patients with common variable hypogammaglobulinaemia. This retrovirus "was clearly related to HTLV-III/LAV" (HIV); evidence included positive western blot with AIDS sera and hybridization with HIV probes.168 In 1984 Montagnier and his colleagues reported that "activated lymphocytes from a healthy blood donor were spontaneously releasing a virus similar to LAV1 in culture."169 With the method presently used for "HIV" isolation (reaction of p24 with proteins in culture) Schupbach and his associates using cultures of whole blood reported positive results in 49/60 (82%) of "presumably uninfected but serologically indeterminate" individuals and in 5/5 (100%) "seronegative blood donors".170
In 1983 Luc Montagnier and his colleagues and in 1984 Robert Gallo and his colleagues claimed to have proven the existence of HIV by purifying retroviral particles, that is, by obtaining a mass of particles isolated from everything else and showing that the particles are infectious. A critical analysis of their evidence shows that neither group presented proof of isolation of a novel retrovirus from AIDS patients. The phenomena they interpreted as HIV are all non-specific and were known to be so long before the AIDS era. In fact, given the origin of the cells and the culture conditions one would be expect to find all these phenomena even if the cultures are not infected with a retrovirus. In 1997 Montagnier himself stressed that to prove the existence of a unique retrovirus purification is absolutely necessary and admitted that he had not presented such proof, and in his view, neither had Gallo. Recognition of these facts may prove the first step in solving the problem of AIDS.
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Competing interests: None declared