The Final Act

SHOULD HIV-AIDS CRITICS QUESTIONTHE EXISTENCE OF HIV?

There is a time in the affairs of men, which, taken at the flood, leads on to fortune; Omitted, all the voyage of their life is bound in shallows and in miseries. On such a full sea are we now afloat; And we must take the current when it serves, or lose our ventures.
-- Julius Caesar , Act V, Scene III

About the authors, biophysicist Eleni Papadopulos-Eleopulos and the phycicians who sit with her on the faculty of the University of Western Australia's medical school.

In recent months AIDS reappraisers have debated the wisdom of taking up the issue of HIV isolation as an argument in our fight against mainstream AIDS science. Those who advise against the isolation discussion list among their reasons:

(i) It will provide HIV/AIDS protagonists additional ammunition with which to discredit us;
(ii) It makes little difference if people are being killed in the name of a non-existent or a merely harmless virus;
(iii) It is an "existentialist" discussion.

From the very beginning (1, 2), we in the Perth group have presented the evidence which may well prove to be the most significant argument against the HIV model of AIDS. If we are correct that the data fail to prove beyond reasonable doubt the existence of HIV then, in terms of a putative exogenous retrovirus, there can be no "observations to provide strong support for the official theory." No one inside or outside the dissident ranks can escape from this point. Although our group has published many scientific and epidemiological reasons for doubting the existence of HIV, we largely avoided making pronouncements such as, "HIV does not exist." We held back for two reasons:

(i) To avoid having our papers rejected by academic journals, since disputing the existence of HIV seems even more outrageous than disputing a causal role for it in AIDS;

(ii) To avoid splitting the beleaguered coalition of HIV-AIDS critics, since its most visible leader, UC-Berkeley biologist Peter Duesberg, steadfastly asserts that the evidence supporting HIV's existence is so overwhelming that disputing it damages the coalition's collective credibility.

We thought we could avoid these problems because we thought we could deconstruct the infectious AIDS model without questioning or refuting the existence of HIV.

WHY AIDS CRITICS MUST DEBATE HIV'S EXISTENCE

In 1996, Peter Duesberg forced our hand. In responding to an of-fer of a cash prize for a paper proving the existence of HIV offered by the British AIDS dissident publication Continuum, his article directly challenged our research (3). Thus, we then had no choice but to openly defend our position (4). We believe now, that despite the political penalties, HIV-AIDS critics must question the isolation -- and thus the existence of -- HIV for the following four reasons:

(i) Since 1996 it has become clear that AIDS dissidents are divided on the isolation issue, and already some of the best known HIV proponents realize it. We can no longer pretend otherwise.

(ii) Even the best formal and direct efforts by Duesberg and even some HIV proponents have failed to identify data that refute our conclusions about HIV's isolation and existence -- the reason Continuum has awarded no one its prize. Though many HIV-AIDS critics (even some who agree with us in private) ignore the isolation issue in their public discourse, they do so only by accepting half-truths.

(iii) "HIV" is the main obstacle -- indeed, the only obstacle -- in deconstructing the HIV model of AIDS.

(iv) Demonstrating that HIV has not been isolated is not an "existentialist" debate. In fact we consider it the strongest argument we can muster.

If AIDS critics accept that no proof establishes the existence of HIV, then the construction "AIDS" -- so premised on the existence of HIV that its proponents call it "HIV disease" -- collapses immediately, and all the so-called "HIV tests" are automatically unmasked as being useless. If, on the other hand, AIDS critics accept the existence of HIV, their debate with HIV-AIDS could continue endlessly, no matter how courageously they fight and no matter how many sacrifices they make. In this regard, Peter Duesberg's unprecedented professional and personal ordeal provides a wise and timely lesson to us all.

WHY HIV ISOLATION IS NECESSARY

The word "isolation" appears frequently in scientific papers and in debate concerning HIV and indeed in virology in general. For example, Montagnier's 1983 and two of Gallo's 1984 Science papers contain the word in their titles as well as the text. Use of this word signals the reader that the paper presents data proving that a virus exists. Authors of the first such report can claim to have discovered the particular virus. Scientific readers must consider whether the data presented as "isolation" do indeed justify this claim.
Thus, the first absolutely necessary, but not sufficient, step in proving the existence of a retrovirus is to isolate retrovirus-like particles. That is, obtain such particles separate from everything else. In other words, purify objects that at least look like retroviruses when viewed by an electron microscope. There are many reasons why isolation is necessary, including the following:

1. To prove that the retrovirus-like particles are infectious, that is, the particles are a virus .
Finding a retrovirus either in vitro or in vivo is not proof that it is exogenous -- that is, it originated from outside the cell -- or that it is infectious. Furthermore, 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." (5) In other words, it is not sufficient to claim a particle is a retrovirus merely on appearances. To prove that retrovirus-like particles observed in a culture are virus particles one must isolate (purify) the particles, characterize their proteins and RNA, and introduce them into a secondary culture, preferably containing cells of a different type than the primary culture. If any particles are released in the secondary culture, isolate them and prove that their proteins and RNA are exactly the same as those of the particles isolated from the primary culture. In these types of experiments, one must not ignore the pivotal importance of proper controls.

2. To determine their biological effects.
For this, only samples of pure virus-like particles will do. Otherwise you cannot determine if the effects result from the possible viruses or from contaminants. As Peter Duesberg has pointed out, Koch's second postulate -- the microbe must be isolated from the host and grown in pure culture -- "was designed to prove that a given disease was caused by a particular germ, rather than by some undetermined mixture of non-infectious substances." Ironically, in 1911 Peyton Rous, the discoverer of retroviruses, issued the same caveat for retroviruses. (6)

3. To characterize the viral proteins.
The only way to prove that a protein is a viral protein is to obtain it from that object or, when the object is very small, as is the case of viruses, from material which contains nothing else but virus particles. If the material contains impurities which also have proteins, it is not possible to determine what is viral and what is not. Only after the viral proteins are characterized they can be used as antigens in the antibody tests.

4. To characterize the viral genome.
As for viral proteins, the only way to prove that a stretch of RNA is viral is to obtain it from material which contains nothing else but virus particles. If the material contains impurities, the impurities must not be constituted from RNA. Then, and only then, can the RNA and its cDNA be used as probes and primers for genomic hybridization and PCR studies.

5. To use it as a gold standard.
Just because a virus or viral protein reacts with an antibody present in a patient's sera, this does not prove that the antibody is directed against the virus or its proteins. That is, that the reaction is specific. To determine the specificity of an antibody reaction, one must use the virus as a gold standard. HIV-AIDS proponents such as Donald Francis, MD, agree. Speculating on a viral cause for AIDS in 1983, Francis wrote, "One must rely on more elaborate detection methods through which, by some specific tool, one can 'see' a virus. Some specific substances, such as antibody or nucleic acids, will identify viruses even if the cells remain alive. The problem here is that such methods 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." [italics ours] (7) The only way to perform hybridization and PCR studies is to use the viral RNA or its cDNA or small fragments of it, as probes and primers. However, as with antibodies which react with viral proteins, a positive result, especially a positive PCR result, does not guarantee that what is detected is viral RNA. To determine the specificity of the PCR, the virus must be used as a gold standard.

HIV "ISOLATION"

All retrovirologists agree that one of the principal defining physical characteristics of retroviruses is their density. In sucrose density gradients they band at a density of 1.16g/ml. Using the method of sucrose density banding in 1983 Francoise Barre-Sinoussi, Luc Montagnier, and their colleagues claimed to have isolated a retrovirus. That is, they claimed to have obtained material which contained nothing else but "purified labeled virus" which now is known as HIV. Robert Gallo's group in 1984 reported similar claims. It goes without saying that if the material constituted pure HIV, then all the proteins present in such material must represent HIV proteins. Instead, they defined as HIV proteins those that they found to react most often with sera from subjects with AIDS or at risk for AIDS. They further defined the reacting antibodies as HIV-specific. Since then, the reaction of these proteins with antibodies has been considered proof of HIV infection. Again, if their material constituted pure HIV, then all the nucleic acids present in their material must represent the HIV genome. Instead, they arbitrarily chose only some fragments of adenine-rich RNA as constituting -- when pieced together -- the HIV genome. (No one has ever claimed to identify a complete HIV genome, which must exist if HIV exists). Since then, these fragments have been used as probes and primers for hybridization and PCR studies, including the determination of "viral load."

The most significant problem in accepting the Montagnier-Gallo claims is the fact that both failed to publish an electron microscope picture of the "pure" HIV. Thus, neither justified their claim of purity, that the material contained nothing else but isolated, retrovirus-like particles, "purified labeled virus." In 1997 Montagnier responded to the French Journalist Djamel Tahi, who asked why his group never published such pictures. Amazingly, Montagnier replied, this was because in what his group called "purified" HIV there were no particles with the "morphology typical of retroviruses." When Tahi asked him if the Gallo group purified HIV, Montagnier replied: "I don't know if he really purified. I don't believe so." (8) If this is the case, then the 1983 Montagnier findings and the 1984 Gallo findings prove beyond all reasonable doubt that they did not have any retrovirus, much less a unique retrovirus, and that the proteins and the RNAs in their "purified" material could not have been of retroviral origin.

In the same year, 1997, the journal Virology published two papers presenting the first electron micrographs of "purified HIV" obtained by banding the supernatant of "infected" cultures in sucrose density gradients (9, 10). The authors of both studies claimed that their "purified" material contained some particles which looked like retroviruses, which they labeled as HIV particles. But they admitted that their material predominantly contained particles which were not viruses but "mock virus", that is, "budding membrane particles frequently called microvesicles."

Furthermore, "The cellular vesicles appear to be a heterogeneous population of both electron-lucent and electron-dense membrane-delineated vesicles ranging in size from about 50 to 500nm." Many but not all of these mock viruses "appear empty by electron microscopy." According to the authors of these studies, one of the reasons that some of the "mock virus" particles appear to have no core "might be that the vesicles contain large amounts of protein and nucleic acid which are unstructured." They showed that the microvesicles "are a major contaminant" of the "purified" HIV. Indeed, the caption to one of the electron micrographs reads, "Purified vesicles from infected H9 cells (a) and activated PBMC (b) supernatants," not purified HIV.

In a further experiment, the supernatant from "non-infected" cultures was also banded in sucrose gradients. The authors claimed that the banded material from these cultures contained only microvesicles, "mock virus" particles but no particles with the morphology of HIV. The mock virus particles contain both DNA and RNA, including poly-A rich mRNA.

No reason is given, other than morphological, for why some of the particles in the fractions from the "infected" cells are virus particles and the others "mock virus." As far as morphology is concerned, none of the particles have all the morphological characteristics attributed to HIV, or even retroviruses. For example, the mean diameters of the particles published by Bess are two and a half times the diameter of any known retrovirus particle.

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.

That the "viral" proteins are nothing more than cellular proteins was further demonstrated by Arthur, Bess and their associates. In their efforts to make an HIV vaccine, they immunized macaques with, amongst other antigens, "mock virus," that is, sucrose density-banded material from the supernatants of non-infected H9 cell cultures. (11) After the initial immunization, 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 immunization 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 immunization, 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 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 hybridization and PCR studies, no matter what results are obtained, they cannot be considered proof for infection with a retrovirus, any retrovirus.

RA Editor Paul Philpott has said, "I think the points that most effectively refute the HIV model have not been taken up as the principal weapons of our most visible advocates." We think the clear failure to demonstrate the existence of HIV represents the most effective point of all.