Distinguishing between true and "official" HIV infection 20 June 2003
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Eleni Papadopulos-Eleopulos,
Biophysicist
Department of Medical Physics, Royal Perth Hospital, Western Australia,
Valendar F Turner, John Papadimitriou, Barry Page, David Causer, Helman Alfonso

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Re: Distinguishing between true and "official" HIV infection

Distinguishing between true and “official” HIV infection

Brian Foley concedes no tests are 100% perfect. Presumably his caveat applies to the HIV antibody tests. This means that before antibody test parameters are verified a scientist must acknowledge the possibility that an unknown proportion of antibody positive patients are not HIV infected. This leads to two questions: what is the proportion and how do we find out? Both are highly significant because it is most undesirable to introduce the test into routine clinical practice before this parameter is ascertained. We will ignore false negatives because physicians and patients most often want to know whether or not an already positive test means HIV infection. Brian Foley correctly identifies at least one reason why a positive test does not necessarily indicate HIV infection. Namely "the patient has elevated levels of some antibody or antibodies that bind to HIV proteins". He then arrives at an explanation of how appearing and disappearing Western blot bands argue the need for "both patients and doctors need to use other data besides the ELISA and WB tests and human intelligence to interpret that data". This is followed by an example to illustrate how "both patients and doctors" might apply themselves to this task. (Do patients really play a role in interpreting their own tests?) The crux of Foley’s interpretation revolves around the time of appearance and the numbers of "weak" or "strong" WB bands. Apparently "two weakly positive western blot bands" may or may not be HIV but "a strong ELISA positive plus 3 or 4 strongly reactive western blot bands" definitely are HIV. Is this system ex cathedra or does Brian have proof? If he has proof what is it? Notwithstanding, is he aware that in the MutiCenter AIDS Cohort Studies, up till 1990 one “strong” WB band was proof of HIV infection? 1 Were “the one strong band” members of this cohort of 5000 gay men “truly HIV-infected” or not? We should also note that Brian’s criteria are yet another set we could add to the list of global criteria for a positive Western blot (see the chart at the end of this posting and in reference 13).

Not surprisingly, patients are about as interested in the arcane and ludicrous distinctions between being "officially" or "unofficially" infected as they are in being "partially pregnant". They just want to know if they’re infected. Is the patient one of the lucky ones whose "elevated levels of some antibody or antibodies that bind to HIV proteins" are not HIV antibodies? Given that AIDS patients are characterised by hypergammaglobulinaemia, that is, they have "elevated levels of some antibody or antibodies", because of antibody cross-reactivities it is more than likely that many and perhaps 100% of the antibodies which "bind to HIV proteins" are not HIV antibodies at all. 2-4 Note this would not negate a correlation between a positive test and AIDS. That is a completely different matter and we trust Brian is cognisant of the distinction. Correlations between diseases and non-specific tests are well recognised and are of great clinical utility. Both in diagnosis, prognosis and predicting the success or failure of treatments. For example, the humble temperature measurement. Or, more attuned with antibody tests, measurements of the erythrocyte sedimentation rate (ESR). 5

As mentioned, Brian’s criteria involve arbitrary appearances and numbers of “weak” and “strong” ELISAs and WB bands. It goes without saying this is not proof of antibody test specificity. If Brian were handed a small object resembling an apple, does giving him a ton of the same but larger three weeks later prove it truly is an apple? The problem with Brian’s, as with all other criteria, is that as real physicians, talking to real patients outside laboratories, we might get stuck with a patient who wants to know the trick of how HIV experts including the CDC and the WHO 6 distinguish between "real" HIV antibodies and the “pretenders”. Obviously one cannot tell merely by inspecting the test tube or paper strip. All we see there is a colour change. Evidence that a reaction has taken place. To cut to the chase and as we have argued elsewhere, 2-4 7-13 the only way to document what proportions of reactions are and are not HIV is to compare the presence or absence of reactions with the presence or absence of HIV itself. That is, against an HIV isolation gold standard. This has never been reported but nonetheless was and always will remain a fundamental requirement.

At the risk of being repetitive, 13 in regard to HIV isolation it is well worth considering what this would mean if an attempt were made to apply Montagnier’s and Gallo’s method to verification. Their method of HIV isolation means this: When cells from AIDS patients are cultured with mitogens and other chemical agents, after a few weeks the cultures (a) reverse transcribe artificial primer-template RNAs; (b) contain a few particles bearing some resemblance to retroviral particles; (c) contain some proteins which react with antibodies present in the serum of AIDS patients. These are the data Montagnier and Gallo presented in Science in 1983/84 as proof of the existence of HIV. Yet (a) reverse transcription is a property of all cells, healthy or unhealthy; 14 15 Normal, mitogenically stimulated “non-HIV-infected” lymphocytes reverse transcribe; 16 17 (b) retroviral-like particles are ubiquitous. 18 As Gallo himself pointed out in 1976, "virus‑like particles morphologically and biochemically [which reverse transcribe] resembling type‑C virus but apparently lacking the ability to replicate, have been frequently observed" in human leukaemias. 19 Particles identical to HIV are found in the majority of normal, human placentas. 20 Particles indistinguishable from HIV are found as frequently (90%) in the enlarged lymph nodes of patients without AIDS or immune deficiency as those with AIDS; 21 (c) as Dr. Foley reminds us, antibodies react with many different substances, not just those which induce their formation. 22 A reaction does not confer “ownership” of either antibody or protein to a unique retrovirus, or indeed to any other object. If this were scientifically valid one could claim that acid curdling milk proves the milk is from a cow and the acid from a lemon. It is precisely because antibodies are not monogamous that their reactions must be verified against an independent gold standard.

In 1997 Montagnier, the discoverer of HIV, admitted he could not find particles “with the morphology typical of retroviruses” in EMs of his “purified virus”. He also stated that to obtain its proteins HIV must be purified but he had not purified HIV. In his opinion neither had Gallo. 11 13 23-25 Neither has anybody else since. Which means that the “HIV” proteins originate from material which has never been proven to contain retroviral particles. Since reverse transcription is a property of all cells and retroviral-like particles are ubiquitous, any attempt to use the Montagnier and Gallo method of isolation to validate the antibody tests would amount to using an antibody test as its own gold standard.

Dr. Foley can inspect the passing parade of ELISAs and Western blot strips from now till eternity but he will not be able to discern which particular antibodies are causing the colour change. All could be due to a retrovirus HIV for whose existence scientific proof may one day be forthcoming. Or none could be due to HIV. Eighteen years after the tests were introduced it is clear, at least in the risk groups, that a positive antibody test is related to AIDS. However, what is very clear is we still don’t know if the positive test is caused by a retroviral infection. It remains inexplicable why scientists have ignored the need to prove the specificity of the HIV antibody tests for HIV infection. If the antibody tests do not prove infection with a retrovirus then a vast amount of the effort expended in combating AIDS is based on an arrant misapprehension. This can only be viewed as a gross waste of resources.

REFERENCES

1. Phair J, Jacobson L, Detals R, Rinaldo C, Saah A, Schrager L, et al. Acquired Immune Deficiency Syndrome Occuring Within 5 Years of Infection with Human Immunodeficiency Virus Type-1: The Multicenter AIDS Cohort Study. Journal of Acquired Immune Deficiency Syndromes 1992;5:490-496

2. Papadopulos-Eleopulos E, Turner VF, Papadimitriou JM. Is a positive Western blot proof of HIV infection? Bio/Technology 1993;11:696-707

3. Papadopulos-Eleopulos E, Turner VF, Papadimitriou JM, Causer D. HIV antibodies: Further questions and a plea for clarification. Current Medical Research and Opinion 1997;13:627-634

4. Papadopulos-Eleopulos E, Turner VF, Papadimitriou JM, Causer D, Page B. HIV antibody tests and viral load - more unanswered questions and a further plea for clarification. Current Medical Research and Opinion 1998;14:185-186

5. Papadopulos-Eleopulos E, Turner VF, Papadimitriou JM, Alfonso H, Page BAP, Causer D, et al. High rates of HIV seropositivity in Africa-alternative explanation. International Journal of STD and AIDS 2003

6. National Institute of Allergy and Infectious Diseases. Focus on the HIV-AIDS Connection, 2001. www.niaid.nih.gov/news room/focuson/hiv00/default.htm

7. Papadopulos E, Turner V. Presentation to the Presidential Panel AIDS Advisory Meeting 3 & 4 July Johannesberg, South Africa. 2000. www.theperthgroup.com/aids/pretoria2.doc

8. Papadopulos-Eleopopulos E, Turner VF, Papadimitriou JM, Hedland-Thomas B, Causer D, Page B. Chart of the global variation in the criteria for a positive HIV Western blot, 1993.www.theperthgroup.com/aids/WBCHART.pdf

9. Papadopulos-Eleopulos E, Turner VF, Papadimitriou JM. Has Gallo proven the role of HIV in AIDS? Emergency Medicine [Australia] 1993;5:113-123

10. Papadopulos-Eleopulos E, Turner VF, Papadimitriou JM, Causer D. The Isolation of HIV: Has it really been achieved? Continuum 1996;4:1s-24s.www.virusmyth.net/aids/data/epreplypd.htm

11. Papadopulos-Eleopulos E. A critical analysis of the evidence for the existence of HIV and the HIV antibody tests: Presentation via satellite to the XIIth International AIDS Conference, Geneva. 1998 (June 28th). www.virusmyth.net.aids/perthgroup/geneva

12. Papadopulos-Eleopulos E, Turner VF, Papadimitriou JM, Alfonso H, Page BAP, Causer D, et al. Mother to Child Transmission of HIV and its Prevention with ATZ and Nevirapine. Perth: The Perth Group, 2001.

13. Papadopulos-Eleopulos E, Turner VF, Papadimitriou JM, Alfonso H, Page BA, Causer D. A critical analysis of the evidence for the existence of HIV. Electronic British Medical Journal 2003. http://bmj.com/cgi/eletters/326/7387/495#31507

14. Varmus H. Reverse Transcription. Scientific American 1987;257:48-54

15. Varmus HE. Reverse transcription in bacteria. Cell 1989;56:721-724

16. Gallo RC, Sarin PS, Wu AM. On the nature of the Nucleic Acids and RNA Dependent DNA Polymerase from RNA Tumor Viruses and Human Cells. In: Silvestri LG, editor. Possible Episomes in Eukaryotes. Amsterdam: North-Holland Publishing Company, 1973:13-34.

17. Tomley FM, Armstrong SJ, Mahy BWJ, Owen LN. Reverse transcriptase activity and particles of retroviral density in cultured canine lymphosarcoma supernatants. British Journal of Cancer 1983;47:277-284

18. Frank H. Retroviridae. In: Nermut MV, Steven AC, editors. Animal Virus and Structure. Oxford: Elsevier, 1987:253-256.

19. Gallo RC, Wong-Staal F, Reitz M, Gallagher RE, Miller N, Gillespie DH. Some evidence for infectious type-C virus in humans. In: Balimore D, Huang AS, Fox CF, editors. Animal Virology. New York: Academic Press Inc., 1976:385-405.

20. Panem S. C Type Virus Expression in the Placenta. Current Topics in Pathology 1979;66:175-189

21. O'Hara CJ, Groopman JE, Federman M. The ultrastructural and immunohistochemical demonstration of viral particles in lymph nodes from human immunodeficiency virus-related and non-human immunodeficiency virus-related lymphadenopathy syndromes. Human Pathology 1988;19:545-9. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi? cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3371979

22. Nossal GJV. Antibodies and Immunity. Harmondsworth, UK: Penguin Books Ltd, 1971.

23. Tahi D. Did Luc Montagnier discover HIV? Text of video interview with Professor Luc Montagnier at the Pasteur Institute July 18th 1997. Continuum 1998;5:30-34. www.virusmyth.net/aids/data/dtinterviewlm.htm

24. Weiss R, Turner, VF. Email debate on the existence of HIV, 1999. www.virusmyth.net/aids/perthgroup/papers2.html

25. Turner VF, Weiss R. Email debate with Professor Robin Weiss on the existence of HIV. 1999. www.theperthgroup.com/aids/vftweiss.html

GLOBAL VARIATION OF THE CRITERIA DEFINING A POSITIVE HIV WESTERN BLOT

AFR=AFRICA;1 AUS=AUSTRALIA;2 FDA=US FOOD AND DRUG ADMINISTRATION;3 RCX=US RED CROSS;3 CDC=US CENTER FOR DISEASE CONTROL;3 CON=US CONSORTIUM FOR RETROVIRUS SEROLOGY STANDARDIZATION;3 GER=GERMANY; UK=UNITED KINGDOM; FRA=FRANCE; MACS= US MULTICENTER AIDS COHORT STUDY 1983-1992. * Bands not in electrophoretic order

NOTES:

I. “The Association of Public Health Laboratories now recommends that patients who have minimal positive results on the WB, eg p24 and gp160 only, or gp41 and gp160 only, be told that these patterns have been seen in persons who are not infected with HIV and that follow-up testing is required to determine actual infective status”.4

II. In February 1993 the US Food and Drug Administration relaxed their criteria in order to “reduce the number of HIV-1 seroindeterminate Western blot interpretations”, that is, to increase the number of HIV positive individuals.5

1. WHO. (1990). Acquired Immunodeficiency Syndrome (AIDS). Proposed criteria for interpreting results from Western blot assays for HIV-1, HIV-2 and HTLV-I/HTLV-II. Weekly Epidemiological Record 65:281-298.

2. Healy DS, Maskill WJ, Howard TS, et al. (1992). HIV-1 Western blot: development and assessment of testing to resolve indeterminate reactivity. AIDS 6:629-633.

3. Lundberg GD. (1988). Serological Diagnosis of Human Immunodeficiency Virus Infection by Western blot Testing. Journal of the American Medical Association 260:674-679. (Data presented in this paper reveal that when the FDA criteria are used to interpret the HIV Western blot less than 50% of US AIDS patients are HIV positive whereas 10% of persons not at risk of AIDS are also positive).

4. Mylonakis E, Paliou M, Greenbough TC, Flaningan TP, Letvin NL, Rich JD. Report of a false-positive HIV test result and the potential use of additional tests in establishing HIV serostatus. Archives of Internal Medicine 2000;160:2386-8.

5. Keinman S, Busch MP, Hall L, et al. (1998). False-positive HIV-1 test results in a low -risk screening setting of voluntary blood donation. Journal of the American Medical Association 280:1080-1083.

Each horizontal band on the left represents a protein with which an antibody can react. Serum from a patient is added to a strip and the strip developed. Where there have been antibodies reacting a coloured band develops. The number and location determining a positive test, for the same virus, varies all over the world.

This gives rise to the incongruity where, for example, an individual positive in New York City on the CDC criteria would not be positive in Sydney, Australia. Or an Australian positive with p41, p32, p24 and p18 bands would not be positive in Africa. Or an African positive with a p41 and p120 band would not be positive in Australia, parts of the US or Europe. Confusion over antibody reactivity is confirmed in diagnostic laboratory manuals. The Genelabs Diagnostic HIV BLOT 2.2 Western blot Assay Instruction Manual advises, “Specific guidelines for interpretation may differ depending on the local policies, GENELABS recommends following the accepted policy to be in accordance with local regulations”. This is followed by seven different criteria for defining a positive Western blot issued by “different international regulatory bodies”. Genelabs also append, “We recommend the following guidelines for the interpretation of the Genelabs Diagnostic HIV BLOT 2.2” and list an eighth set of criteria for a positive Western blot (Packet Insert, 1998). This means that “different international regulatory bodies” or “local policies”, and not the presumed pathogen determine patterns of antibody reactivity said to prove a retroviral infection. Manufacturer Bio-Rad advises “Each laboratory performing Western blot testing should develop its own criteria for band interpretation. Alternatively, band interpretation may be left to the clinician" (Bio-Rad Laboratory Manual 1993).

Competing interests: None declared