Date: Thu, 9 Mar 1995 14:05:46 -0600
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From: sma at wubios.wustl.edu
Subject: Does HIV exist?
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diseases. Even if the virus could have been isolated from all patients,
given the nature of retroviruses and the method used for HTLV-III isolation
(cultures, mitogenic stimulation, co- cultivation) the possibility cannot be
excluded that the virus did not exist in vivo (in AIDS patients), and that
it was a provirus whose expression was facilitated by the culture
conditions. The only method used to prove HIV infection in vivo was the
antibody tests. Such a test can only be used only after its specificity has
been proven by use of the only possible gold standard, the virus itself.
This has not been done.
Furthermore, the antibody test used by Gallo was ELISA, at present known to
be non-reproducible and non-specific. In a study of 1.2 million healthy
military applicants conducted by Colonel Donald Burke and his colleagues,63
it was found that although approximately 1% of all individuals had an
initial positive HIV ELISA, only 50% of repeat ELISAs were positive. Of the
latter, only approximately one third were associated with two subsequent
positive WBs. In Russia, in 1990, out of 20,000 positive ELISAs "only 112
were confirmed" using the WB as a gold standard. In 1991, of approximately
30,000 positive ELISAs, only 66 were confirmed.64
Nowhere in the four Science papers was HTVL-III cytotoxicity mentioned. The
only reference to any cellular abnormalities or pathology in general is in
the first paper where one reads: "The virus positive cultures consistently
showed a high proportion of round giant cells containing numerous nuclei
(Fig. 1a). These cells resemble those induced by HTLV-I and -II except that
the nuclei exhibit a characteristic ring formation". (Fig. 1a is a "light
microscopic examination of clone H4/HTLV-III"). The H4 clone was obtained
from the HT cell line "using irradiated mononuclear cells from peripheral
blood of a healthy blood donor as a feeder". At present, it is known that
the HT cell line and thus H4 are HUT78, derived in 1980 from a patient with
mature T4-cell leukaemia,65,66 However, other cell lines derived from
patients with the same clinical syndrome are known to exhibit similar
morphologies including multinucleated giant cells.67 Thus the cellular
morphological characteristics observed in the first paper may have been an
intrinsic property of the HT cell line, or the result of the culture
conditions, or both, and not due to HTLV-III.
Finally, Gallo and his colleagues did not provide any data on the
immunological status of those individuals from whom viral isolation was
attempted, and no data was presented proving that:
1. HTLV-III (HIV) is both a necessary and sufficient cause of T4- cell
2. T4-cell depletion is both necessary and sufficient for the appearance of
the AIDS indicator diseases.
CONCLUSIONS. The data and arguments that have been presented by Gallo and
his colleagues do not constitute proof of HIV isolation or an unambiguous
role for HIV in the pathogenesis of AIDS. Although some researchers
currently use methods of "viral isolation" essentially the same as that
described by Gallo's group, most use less rigorous methods including
singleton detection of p24 (by antibody techniques), or RT. Notwithstanding,
with all of these techniques, including that described by Gallo and his
colleagues, which itself seen to be greatly problematic, HIV cannot be
"isolated" from 20%-70% of HIV positive and AIDS patients.68,69 Thus we are
faced with a problem of considerable importance. The HIV antibody tests,
both ELISA and WB, the only routinely used tests proving the existence in
vivo of HIV, have yet to be verified against the only suitable gold
standard, viral isolation. The available evidence suggests that this long
overdue but most basic requirement of test evaluation is likely to prove an
immense problem, and while the HIV antibody tests are useful prognostic
markers in the high risk groups, their use as diagnostic and epidemiological
tools for HIV infection is questionable.
Footnote 1. In the Western Blot test, proteins are electrophoretically
separated according to molecular weight and charge. The separated proteins
are then transferred on to nitrocellulose strips by a process known as
electroblotting. When sera are added and the strips developed, coloured
bands appear representing sites of protein/antibody reactions. Each band is
designated by a small "p" for the protein followed by its molecular weight
Footnote 2. In the ELISA (Enzyme Linked Immunosorbent Assay), unseparated
proteins are attached to a solid base such as the walls of plastic tubes or
microplates. The serum being tested is incubated in these containers where
antibody is fixed to the solid phase antigens. After washing,
enzyme-labelled anti-human immunoglobulin is added and also incubated. The
containers are again washed and a substrate specific for the enzyme is
introduced. The resulting colour change is proportional to the amount of
antibody present and is read by eye, or with a spectrophotometer.
ACKNOWLEDGEMENTS We wish to thank all our colleagues and especially Udo
Sch=81klenk, Barry Page, Bruce Hedland-Thomas, David Causer, Richard Fox,
John Peacock, David Prentice, Ronald Hirsch, Patricia Shalala, Keith Jones,
Alun Dufty, June Rider Jones, Coronary Barrow, Dorothy Davis, Julian Smith,
Mark Strahan, Vincent Turner, Wallace Turner, Gary James and Graham Drabble
for their continued support and assistance.
1. Essex M, McLane MF, Lee TH, et al. Antibodies to Cell Membrane Antigens
Associated with Human T-Cell Leukemia Virus in Patients with AIDS. Science
2. Gallo RC, Sarin PS, Gelmann EP, et al. Isolation of Human T- Cell
Leukemia Virus in Acquired Immune Deficiency Syndrome (AIDS). Science
3. Gallo RC. The First Human Retrovirus. Sci Am 1986; 255:78-88.
4. Marx JL. Human T-Cell Leukemia Linked to AIDS. Science 1983;220:806-809.
5. Barr=82-Sinoussi F, Chermann JC, Rey F, et al. Isolation of a
T-Lymphotrophic Retrovirus from a patient at Risk for Acquired Immune
Deficiency Syndrome (AIDS). Science 1983;220:868-871.
6. Popovic M, Sarngadharan MG, Read E, et al. Detection, Isolation,and
Continuous Production of Cytopathic Retroviruses (HTLV-III) from Patients
with AIDS and Pre-AIDS. Science 1984;224:497-500.
7.Gallo RC, Salahuddin SZ, Popovic M, et al. Frequent Detection and
Isolation of Cytopathic Retroviruses (HTLV-III) from Patients with AIDS and
at Risk for AIDS. Science 1984;224:500-502.
8. Sch=81pbach J, Popovic M, Gilden RV, et al. Serological analysis of a
Subgroup of Human T-Lymphotrophic Retroviruses (HTLV-III) Associated with
AIDS. Science 1984;224:503-505.
9. Sarngadharan MG, Popovic M,Bruch L, et al. Antibodies Reactive to Human
T-Lymphotrophic Retroviruses (HTLV-III) in the Serum of Patients with AIDS.
10. Culliton BJ. Gallo Inquiry Takes Puzzling New Turn. Science
11. Culliton BJ. Inside the Gallo Probe. Science 1990;248:1494-1498.
12. Hamilton DP. What Next in the Gallo Case? Science 1991;254:944-945.
13. Palca J. Draft of Gallo Report Sees the Light of Day. Science
14. Cohen J. HHS: Gallo Guilty of Misconduct. Science 1993:259:168-170.
15. Gallo RC, Sarin PS, Kramarsky B. et al. First isolation of HTLV-III.
16. Rous P. A Sarcoma of the Fowl transmissible by an agent separable from
the Tumor Cells. J Exp Med 1911;13:397-411.
17. Weiss R,Teich N, Varmus H, Coffin J. RNA Tumor Viruses. Cold Spring
Harbor Laboratory. Cold Spring Harbor, New York;1982.
18. Temin HM, Baltimore D. RNA-Directed DNA Synthesis and RNA Tumor Viruses.
Adv Vir Res 1972;17:129-186.
19. Toplin I. Tumor Virus Purification using Zonal Rotors. Spectra 1973;No.
20. Bader JP. Reproduction of RNA Tumor Viruses. In: Fraenkel-Conrat H,
Wagner RR, eds. Comprehensive Virology Vol.4. New York: Plenum Press,
21. Sinoussi F, Mendiola L, Chermann, JC, et al. Purification and partial
differentiation of the particles of murine sarcoma virus (M. MSV) according
to their sedimentation rates in sucrose density gradients. Spectra 1973;No.
22. Maddox J. More on Gallo and Popovic. Nature 1992;357:107-109.
23. Lee MH, Sano K, Morales FE, et al. Sensitive Reverse Transcriptase Assay
to Detect and Quantitate Human Immunodeficiency Virus. J Clin Microb
24. 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, ed. Possible Episomes in Eukaryotes. Amsterdam: North-Holland
Publishing Company, 1973:13-34.
25. Whitkin SS, Higgins PJ, Bendich A. Inhibition of reverse transcriptase
and human sperm DNA polymerase by anti-sperm antibodies. Clin Exp Immunol
26. Varmus H. Reverse Transcription Sci Am 1987;257:48-54.
27. Hart DJ, Gogu SR, Agrawal KC, et al. Inhibition of RNA-dependent DNA
polymerases (reverse transcriptase) of normal cells by activated
azidothymidine:a possible basis for drug toxicities? In: Vol I, Abstracts
VII International Conference on AIDS,Florence, 1991:110.
28. Gallo RC, Wong-Staal F, Reitz M, et al. Some Evidence For Infectious
Type-C Virus in Humans. In: Baltimore D, Huang AS, Fox CF, eds. Animal
Virology. New York: Academic Press Inc.,1976:385-407.
29. Panem S, Prochownik EV, Reale FR, et al. Isolation of Type C Virions
from a Normal Human Fibroblast Strain. Science 1975;189:297-299.
30. Panem S, Prochownik EV, Knish WM, Kirsten WH. Cell Generation and Type-C
Virus Expression in the Human Embryonic Cell Strain HEL-12. J Gen Virol
31. Panem S. C Type Virus Expression in the Placenta. Curr Top Pathol
32. Sarngadharan MG, Robert-Guroff M, Gallo RC. DNA Polymerases of Normal
and Neoplastic Mammalian Cells. Biochim Biophys Acta 1978;516:419-487.
33. Klatzmann D, Barr=82-Sinoussi F, Nugeyre MT, et al. Selective Tropism of
Lymphadenopathy Associated Virus (LAV) for Helper-Inducer T Lymphocytes.
34. Montagnier L. Lymphadenopathy-Associated Virus:From Molecular Biology to
Pathogenicity. Ann Int Med 1985;103:689-693.
35. Gendelman HE, Orenstein JM, Martin MA, et al. Efficient Isolation and
Propagation of Human Immunodeficiency Virus on Recombinant
Colony-Stimulating Factor 1-Treated Monocytes. J Exp Med 1988;167:1428-1441.
36. Gelderblom HR, =99zel M, Hausmann EHS, et al. Fine Structure of Human
Immunodeficiency Virus (HIV), Immunolocalization of Structural Proteins and
Virus-Cell Relation. Micron Microsc 1988;19:41-60.
37. Lecatsas G, Taylor MB. Pleomorphism in HTLV-III, the AIDS virus. S Afr
Med J 1986;69:793-794.
38. Hockley DJ, Wood RD, Jacobs JP, et al. Electron Microscopy of Human
Immunodeficiency Virus. J Gen Virol 1988;69:2455-2469.
39. Dourmashkin RR, O'Toole CM, Bucher D, Oxford JS. The presence of budding
virus-like particles in human lymphoid cells used for HIV cultivation. In:
Vol I, Abstracts VII International Conference on AIDS,Florence,1991:122.
40. O'Hara CJ, Groopmen JE, Federman M. The Ultrastructural and
Immunohistochemical Demonstration of Viral Particles in Lymph Nodes from
Human Immunodeficiency Virus-Related Lymphadenopathy Syndromes. Hum Pathol
41. Henderson LE, Sowder R, Copeland TD, et al. Direct Identification of
Class II Histocompatibility DR Proteins in Preparations of Human T-Cell
Lymphotropic Virus Type III. J Virol 1987;61:629-632.
42. Lundberg GD. Serological Diagnosis of Human Immunodeficiency Virus
Infection by Western Blot Testing. JAMA 1988;260:674-679.
43. Edwards VM, Mosley JW and the Transfusion Safety Study Group.
Reproducibility in Quality Control of Protein (Western) Immunoblot Assay for
Antibodies to Human Immunodeficiency Virus. Am J Clin Pathol 1991;91:75-78.
44. Pinter A, Honnen WJ, Tilley SA, et al. Oligomeric Structure of gp41, the
Transmembrane Protein of Human Immunodeficiency Virus Type 1. J Virol
45. Zolla-Pazner S, Gorny MK, Honnen WJ. Reinterpretation of Human
Immunodeficiency Virus Western Blot Patterns. NEJM 1989;320:1280-1281.
46.Damsky CH, Sheffield JB, Tuszynski GP, et al. Is there a role for Actin
in Virus Budding? J Cell Biol 1977;75:593-605. 47. Stanislawsky L, Mongiat
F, Neto VM, et al. Presence of Actin in Oncornaviruses. Biochem Biophys Res
48. Stricker RB, Abrams DI, Corash L, et al. Target Platelet Antigen in
Homosexual Men with Immune Thrombocytopenia. NEJM 1985;313:1375-1380.
49. Matsiota P, Chamaret S, Montagnier L. et al. Detection of Natural
Autoantibodies in the serum of Anti-HIV Positive-Individuals. Ann Inst
50. Wong-Staal F, Gallo RC. Human T-lymphotropic retroviruses. Nature
51. Genesca J, Jett BW, Epstein JS, et al. What do Western Blot
indeterminate patterns for Human Immunodeficiency Virus mean in EIA-negative
blood donors? Lancet 1989;II:1023-1025. 52. Ranki A, Johansson E, Krohn K.
Interpretation of Antibodies Reacting Solely with Human Retroviral Core
Proteins. NEJM 1988;318:448-449.
53. Delord B, Ottmann M, Schrive MH, et al. HIV-1 expression in 25 infected
patients:A comparison of RNA PCR, p24 EIA in Plasma and in situ
Hybridization in mononuclear cells. In: Vol. I, Abstracts VII International
Conference on AIDS,Florence, 1991:113.
54. Todak G, Klein E, Lange M, et al. A clinical appraisal of the p24
Antigen test. In:Vol. I, Abstracts VII International Conference on
55. Mager DL, Freeman JD. Human Endogenous Retroviruslike Genome with Type C
pol Sequences and gag Sequences Related to Human T-Cell Lymphotropic
Viruses. J Virol 1987;61:4060-4066. 56. Banki K, Maceda J Hurley E, et al.
Human T-cell lymphotropic virus (HTLV)-related endogenous sequence, HRES-1
encodes a 28-kDa protein: A possible autoantigen for HTLV-I gag- reactive
autoantibodies. Proc Natl Acad Sci 1992;89:1939-1943.
57. Horwitz MS, Boyce-Jacino MT, Faras AJ. Novel Human Endogenous Sequences
Related to Human Immunodeficiency Virus Type 1. J Virol 1992;66:2170-2179.
58. Aaronson SA, Todaro GJ, Scholnick EM. Induction of murine C- type
viruses from clonal lines of virus-free BALB/3T3 cells. Science
59. Hirsch MS, Phillips SM, Solnik C, et al. Activation of Leukemia Viruses
by Graft-Versus-Host and Mixed Lymphocyte Reactions In Vitro. Proc Nat Acad
60. Toyoshima K, Vogt PK. Enhancement and Inhibition of Avian Sarcoma
Viruses by Polycations and Polyanions. Virol 1969;38:414-426.
61. Todaro GJ, Benveniste RE, Sherr CJ. Interspecies Transfer of RNA Tumour
Virus Genes: Implications for the search for "Human" Type C Viruses. In:
Baltimore D, Huang AS, Fox CS, eds. Animal Virology. New York: Academic
62. Connor S. AIDS: Science Stands On Trial. New Scientist 1987; Feb
63. Burke DS, Brundage JF, Redfield RR, et al. Measurement of the False
Positive Rate in a Screening Program for Human Immunodeficiency Virus
Infections. NEJM 1988;319:961-964.
64. Voevodin A. HIV screening in Russia. Lancet 1992;339:1548.
65. Gazdar AF, Carney DN, Bunn PA, et al. Mitogen Requirements for the In
Vitro Propagation of Cutaneous T-Cell Lymphomas. Blood 1980;55:409-417.
66. Rubinstein E. The Untold Story of HUT78. Science 1990;248:1499-1507.
67. Poiesz B, Ruscetti FW, Mier JW, et al. T-cell lines established from
human T-lymphocytic neoplasias by direct response to T-cell growth factor.
Proc Natl Acad Sci 1980;77:6815-6819.
68. Chiodi F, Albert J, Olausson E, et al. Isolation Frequency of Human
Immunodeficiency Virus from Cerebrospinal Fluid and Blood of Patients with
Varying Severity of HIV Infection. AIDS Res Hum Retrovirol 1988;4:351-358.
69. Learmont J, Tindall B, Evans L, et al. Long-term symptomless HIV-1
infection in recipients of blood products from a single donor. Lancet