<citation>Papadopulos-Eleopulos et al comment on the "seminal" Gallo papers:

By Eleni Papadopulos-Eleopulos, Valendar F. Turner, John M. Papadimitriou
Excerpt from an unpublished draft.

Original Publication
HEAL Toronto

For a published examination of these "seminal" papers see:
HAS GALLO PROVEN THE ROLE OF HIV IN AIDS?
Has Gallo Proven the Role of HIV in AIDS?
Eleni Papadopulos-Eleopulos, Valendar F. Turner, John M. Papadimitriou
Emergency Medicine 1993;5:5-147

... in May 1984, Gallo, Popovic and their colleagues published four papers in Science in which they claimed to have isolated another retrovirus from AIDS patients. The Gallo virus was called human lymphotropic virus-III (HTLV-III). On the 23rd of April 1984, at a Washington press conference held two weeks before the Science papers were published, Margaret Heckler, the then Health and Human Services Secretary, announced that Gallo and his co-workers had discovered the "probable" cause of AIDS and had developed a sensitive test to show whether the "AIDS virus" is present in blood. In 1985, the Pasteur Institute alleged that Gallo had misappropriated LAV in developing the blood test. The ensuing conflict, which reached the American courts, was eventually settled by a negotiated agreement signed in 1987 by Gallo, Montagnier, US President Reagan and French Premier Chirac. The agreement declared Gallo and Montagnier to be co-discoverers of the AIDS virus, presently known as the Human Immunodeficiency Virus. Nevertheless, the misappropriation conflict drew the attention of John Crewdson, an investigative journalist, and US Senator John Dingell. In November 1989, Crewdson published a lengthy article in the Chicago Tribune newspaper, "With allegations that Robert C. Gallo stole from French scientists the virus he discovered to be the cause of AIDS". (30) This led to a National Institute of Health (NIH) internal "inquiry" into the allegation with "an outside committee of expert but disinterested parties [led by Yale biochemist Frederic Richards] to oversee the activity of the internal panel". (31) Following the inquiry, which was viewed as a fact-finding mission, the Richards committee insisted on a "formal investigation...on suspect data in one of four seminal papers published by Gallo's lab in Science on 4 May 1984". (32) In the first paper, with Mikulas Popovic the principal author, "there appears to be differences between what was described in the paper and what was done". (30) A draft report of the formal investigation written by NIH Office of Scientific Integrity (OSI), was published in September 1991. In the draft report, Popovic is accused "of misconduct for misstatements and inaccuracies" that appeared in the paper, and that Gallo, as laboratory chief, "created and fostered conditions that give rise to falsified/ fabricated data and falsified reports". However, Gallo's actions were not considered to "meet the formal definition of misconduct". (33) The final draft report of the OSI, completed in January 1992, was immediately criticised by the Richards Panel as well as Senator Dingell. This led to a review of the OSI report by the Office of Research Integrity (ORI), which found Gallo guilty of scientific misconduct. Nonetheless, the scientific misconduct is said not to "negate the central findings of the [1984 Science] paper". (33-34)

In this first paper entitled "Detection, Isolation and Continuous Production of Cytopathic Retroviruses (HTLV-III) from Patients with AIDS and pre-AIDS", experiments were described in which "concentrated culture fluids harvested from short-term cultures of T-cells" from patients with AIDS or pre-AIDS were cultured with highly selected clones of a leukaemic cell line (HT). The data presented as proof of isolation of HIV were (i) RT activity in cell free supernatants and concentrated culture fluids after "purification" through a sucrose density gradient; (ii) immunofluoroescence of cells obtained by reactions with "Rabbit antiserum to HTLV-III" and "Patient serum (E.T.)"; (iii) electron microscopy showing the presence of extracellular particles. Reading this paper the impression is gained that the HT cell line was cultured with concentrated (supernatant) fluids originating from individual, AIDS patient, mitogen stimulated T-cell cultures. However, the OSI enquiry found that the HT cell line was cultured with concentrated fluids pooled initially from individual cultures of three patients and ultimately from the individual cultures of ten patients. (35) In evidence given to the 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 sufficient proof that the initial, individual specimens contained a retrovirus before they were mixed and added to the HT cell line. The Gallo investigation found the pooling of specimens to be "of dubious scientific rigor". One scientist described the procedure as "really crazy". (31) In essence it is no different from investigating an outbreak of a new type of pneumonia by culturing the sputum of each patient and, when no organisms are cultured, combining all specimens and reculturing.

In the second paper Gallo and his colleagues claimed to have "isolated" HTVLV-III (HIV) from 26/72 (36%) of AIDS patients. The explanation for the low yield of virus isolation was, "many tissue specimens were not received or handled under what we now recognize as optimal conditions. This is particularly so for the samples received from late-stage AIDS patients. Such samples usually contain many dying cells". However, Gallo and his colleagues were widely regarded as world leading experts in culturing tissues for retroviruses and it is now claimed that in AIDS patients who have not received antiviral treatment, "plasma viral RNA levels...at baseline ranged from 10 (4.6) to 10 (7.2) molecules per ml". (36) (equivalent to 10 (4.2) to 10 (6.9) [approximately one hundred thousand to ten million] "virions" per ml (37) ). In this paper the criteria listed for the isolation of HTLV-III were "more than one of the following": "repeated detection of a Mg (2+) -dependent reverse transcriptase activity in supernatant fluids; virus observed [in the cultures, not at the density of 1.16 gm/ml] by electron microscopy (EM); intracellular expression of virus-related antigens detected with antibodies from seropositive donors or with rabbit antiserum to HTLV-III; or transmission of particles". By transmission of particles was meant detection of RT or particles in cultures of human umbilical cord blood, bone marrow or peripheral blood T- lymphocytes, cultured with supernatants from the "infected" cultures. (38) (It can be seen that the Gallo group method permitted instances of "isolation" of a retrovirus without the necessity of evidence for either particles or RT activity). In the third paper it was reported that from the supernatant of the "infected" cultures which, in sucrose density gradients banded at 1.16 gm/ml, two proteins, p41/45 and p24/25, reacted with various human 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". (Of interest is the fact that Montagnier still regards p41/45 as the ubiquitous cellular protein actin and not an HIV protein. (25, 39) (Later, a number of other proteins present in either the material from culture supernatants banding at 1.16 gm/ml or just cellular extracts, and for no reason other than each reacted with antibodies present in sera from AIDS patients or those are risk of developing AIDS, were considered to be specific, HIV 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 are not Lentiviruses). From their data Gallo and his colleagues claimed "isolation of cytopathic retroviruses (HTLV-III) from patients with AIDS" and that their data "provide strong evidence of a causative involvement of the virus in 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". (40)