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    Scientific Papers > Duesberg Defends Challenges.... Part 1


Duesberg Defends Challenges to the Existence of HIV:
Article 1 of 2 for Continuum

-adapted from Continuum, vol 4 (2) pages 8-9, July/August 1996

In 1983, Montagnier et al. have isolated a retrovirus, now termed Human immunodeficiency virus (HIV), from a patient with lymphadenopathy and proposed that HIV may cause AIDS. Antibody against this virus has since been found in many, but not all AIDS patients (Duesberg, 1993) and in 17 million healthy people (World Health Organization, 1995).

Eleni Papadopulos, Val Turner, John Papadimitriou, David Causer, Bruce Hedland-Thomas & Barry Page (Papadopulos-Eleopulos et al., 1995) and Stefan Lanka (Lanka, 1995) maintain that the very existence of HIV is dubious because (i) HIV has not been properly isolated and thus could not have been properly identified. According to Papadopulos et al. "HIV has never been isolated as an independent particle separate from everything else." (Papadopulos-Eleopulos et al., 1995) ; and (ii) because antibodies against HIV are not specific (Papadopulos-Eleopulos, Turner and Papadimitriou, 1993) . They submit that the following evidence is not "specific" for HIV: identifying in the growth medium of infected human cell cultures either the existence of virus-like particles with the electron microscope, or of reverse transcriptase associated with such particles, or of certain HIV antigens or proteins associated with particles, because each of these could be cellular materials or could be from cell-borne (endogenous) retroviruses other than HIV. Indeed, each of these criteria could reflect another retrovirus, and some of theses criteria, eg. particels and proteins, could reflect non-viral material altogether.

However, the Papadopulos-Lanka challenge, that HIV does not exist, fails to explain (i) why virtually all people who contain HIV DNA also contain antibodies against Montagnier"s HIV strain -the global standard of all "HIV tests"- and (ii) why most, but certainly not all people who lack HIV DNA contain no such antibodies. The presence of HIV-reactive antibodies in some uninfected people reflects an inherent limitation of tests for antibodies against viruses and other microbes. Since even the simplest microbes display thousands of antibody docking sites, termed epitopes, antibodies against a given microbe may cross-react with an otherwise unrelated microbe if the two share some epitopes.

In view of the current controversy about the identification of HIV, the British AIDS magazine Continuum has offered in its Jan./Feb. issue a "Missing virus! 1000 Reward" for prove of the isolation of HIV, and the reward was reposted "105 days later ... we"re still waiting" in the March/April, 1996 issue (Christie, 1996a; Christie, 1996b; Continuum, 1996) . The stakes have since been raised considerably by a private reward of 10,000 from Alex Russell from The DMS Watson Library, University College London (letter from Alex Russell to Peter Duesberg, 2/28/96), and have now been raised even further by a 25,000 reward from James Whitehead from The International AIDS Freedom Network (IAFN), London (letter from Fred Cline, San Francisco Representative of the IAFN, April 2, 1996; and letter from Alex Russell to Fred Cline, undated, April 1996).

Here I take up these challenges. I will argue that HIV exists, and has been properly identified as a unique retrovirus on the grounds that (i) it has been isolated - even from its own virion structure - in the form of an infectious, molecularly cloned HIV DNA that is able to induce the synthesis of a reverse transcriptase containing virion, and (ii) that HIV-specific, viral DNA can be identified only in infected, but not in uninfected human cells. I will base my case for the isolation of HIV on the most rigorous method available to date, ie. molecular cloning of infectious HIV DNA, rather than only on the much less stringent, traditional "rules for isolation of a retrovirus ... discussed at the Pasteur Institute, Paris, in 1973" that were stated criteria of isolation in the Continuum"s Missing virus reward (Continuum, 1996) . Indeed I will show that molecular cloning of infectious HIV DNA exceeds the criteria of the old "Pasteur rules".

(i) Isolation of HIV: The existence of the retrovirus HIV predicts that HIV DNA can be isolated from the chromosomal DNA of infected cells. This prediction has been confirmed as follows: Full-length HIV-1 and HIV-2 DNAs have been prepared from virus-infected cells and cloned in bacterial plasmids (Fisher et al., 1985; Levy et al., 1986; Barnett et al., 1993) . Such clones are totally free of all viral and cellular proteins, and cellular contaminants that copurify with virus banded in sucrose gradients at a density of 1.16 g/ml according to the "Pasteur rules". Indeed, these clones are even free of genomic HIV RNA. Infectious HIV-1 and HIV-2 DNA clones productively infect human cells to initiate HIV replication (Fisher et al., 1985; Levy et al., 1986; Barnett et al., 1993) . Such infected ("transfected") cells contain HIV-specific DNA, and produce particles that contain reverse transcriptase, HIV-specific antigens (Fisher et al., 1985; Levy et al., 1986) , have diameters of 100 nm under the elctron microscope (Fisher et al., 1985) , as expected for retroviruses, and produce infectious virus that is neutralized by antisera from certain AIDS patients (Fisher et al., 1985; Levy et al., 1986; Barnett et al., 1993) . Since infectious HIV DNA has been isolated from infected human cells that is free of HIV"s own proteins and RNA as well as from all cellular macromolecules, HIV isolation has passed the most vigorous standards available today. In other words these infectious DNA clones meet and exceed the isolation standards of the traditional "Pasteur rules". Isolation of infectious HIV DNAs is theoretically the most absolute form of isolation - it is the equivalent of isolating the virus" soul, its genetic code, from the virus" body, the virus particle. Thus HIV isolation based on molecular cloning exceeds the old standards defined as "Pasteur rules" by Continuum.

(ii) Identification of HIV. The existence of HIV predicts that infected cells contain a unique, virus-specific DNA of 9150 nucleotides that cannot be detected in DNA of uninfected human cells. The probabilities that cellular DNA and other viral DNAs would contain the same sequence of 9150 nucleotides is 1 in 49150, or 1 in 104500 - extremly close to zero!
Because of the outrageous interest in HIV as the hypothetical cause of AIDS, many investigators have sought specific HIV DNA in humans with and without AIDS in an effort to confirm the rather unreliable HIV antibody-test (Duesberg, 1993; Papadopulos-Eleopulos, Turner and Papadimitriou, 1993).

Because only 1 in 100 T-cells are ever infected in humans, virtually all such studies use Kary Mullis" polymerase chain reaction, a technique that is designed to amplify a DNA-needle into an DNA-haystack. Such efforts have confirmed the existence of HIV-specific DNA in most (not all) antibody-positive persons with and without AIDS - but not in the DNA of antibody-negative people. For example Jackson et al. have tested blood cells of 409 antibody-positives including 144 AIDS patients and 265 healthy people. In addition 131 antibody-negatives were tested. HIV-specific DNA subsets -defined in size and sequence by HIV-specific primers (start signals for the selective amplification) - were found in 403 of the 409 antibody-positives, but in none of the 131 antibody-negative people (Jackson et al., 1990).

In conclusion: HIV has been isolated by the most rigorous method science has to offer. An infectious DNA of 9.15 kilo bases (kb) has been cloned from the cells of HIV-antibody-positive persons, that -upon transfection- induces the synthesis of an unique retrovirus. This DNA "isolates" HIV from all cellular molecules, even from viral proteins and RNA. Having cloned infectious DNA of HIV is as much isolation of HIV as one can possibly get, it is like isolating the fifth symphony from an orchestra hall by recording it on a CD. The retrovirus encoded by this infectious DNA reacts with the same antibodies that crossreact with Montagnier"s global HIV standard, produced by immortal cell lines in many labs and companies around the world for the HIV-test. This confirms the existence of the retrovirus HIV.

The uniqueness of HIV is confirmed by the detection of HIV-specific DNA sequences in the DNA of most antibody-positive people. The same DNA is not found in uninfected humans, and the probability to find such a sequence in any DNA sample is 1/49500 - which is much less likely than to encounter the same water molecule twice by swimming in the Pacific ocean every day of your life.

The existence of an unique retrovirus HIV provides a plausible explanation for the good (not perfect) correlation between the existence of HIV DNA and antibodies against it in thousands of people that have been subjected to both tests. The Papadopulos-Lanka challenge fails to explain this correlation.

Ergo: The Papadopulos-Lanka challenge is rejected. HIV exists and has been isolated. Therefore I gratefully accept the rewards posted for providing the evidence that HIV exists, and was isolated from Continuum, Alex Russell and James Whitehead.

Barnett, S. W., M. Quiroga, A. Werner, D. Dina and J. A. Levy, 1993. Distinguishing features of an infectious molecular clone of the highly divergent and noncytopathic human immunodeficiency virus type 2 UC1 strain. J. Virol. 67: 1006-1014.
Christie, H., 1996a. 105 days ... we"re still waiting. Continuum, March/April, p5.
Christie, H., 1996b. Letter to Peter Duesberg.
Continuum, 1996. The Jody Wells Memorial Prize; Missing Virus! 1,000 Reward. Continuum 3: ?
Duesberg, P. H., 1993. The HIV gap in national AIDS statistics. Biotechnology 11: 955-956.
Fisher, A. G., E. Collalti, L. Ratner, R. C. Gallo and F. Wong-Staal, 1985. A molecular clone of HTLV-III with biological activity. Nature (London) 316: 262-265.
Jackson, J. B., S. Y. Kwok, J. J. Sninsky, J. S. Hopsicker, K. J. Sannerud, F. S. Rhame, K. Henry, M. Simpson and H. H. Balfour Jr., 1990. Human immunodeficiency virus type 1 detected in all seropositive symptomatic and asymptomatic individuals. J. Clin. Microbiol. 28: 16-19.
Lanka, S., 1995. HIV reality or artefact? Continuum, April/May, p4-9.
Levy, J. A., C. Cheng-Mayer, D. Dina and P. A. Luciw, 1986. AIDS retrovirus (ARV-2) clone replicates in transfected human and animal fibroblasts. Science 232: 998-1001.
Papadopulos-Eleopulos, E., V. F. Turner and J. M. Papadimitriou, 1993. Is a positive Western blot proof of HIV infection? Biotechnology 11: 696-707.
Papadopulos-Eleopulos, E., V. F. Turner, J. M. Papadimitriou, D. Causer, B. Hedland-Thomas and B. A. P. Page, 1995. A critical analysis of the HIV-T4-cell-AIDS hypothesis. Genetica 95: 5-24.
World Health Organization, 1995. The current Global Situation of the HIV/AIDS Pandemic. Geneva, Jan.
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