Department of Medical Physics, Royal Perth Hospital, Western Australia, 6001,
Valendar F Turner, John Papadimitriou, Barry Page, David Causer, Helman Alfonso, Sam Mhlongo, Todd Miller, Christian Fiala
Send response to journal:
IS "HIV" SUFFICIENT OR NECESSARY FOR T4 DECREASE (AID)?
In his second rapid response entitled" "Re: Montagnier's reverse transcriptase activity", 26th July, Nicholas Bennett wrote: "Further to my previous reply, at least one of the references directly contradicts the Perth Group on several accounts (as usual):
Zagury, D., Bernard, J., Leonard, R et al (1986), Long term cultures of HTLV-III infected cells: a model of cytopathology of T-cell depletion in AIDS. Science, 231, 850-853.
Not only states that control cultures were done using uninfected cells, but that these cells did not display the same antigens as those infected with HIV, and the "oxidative stress" used was in fact a commonly used mitogen called phytohemaglutinin. Hardly a stress. Infected cells, once activated, died due to the effects of the infection. Uninfected cells did not.
If I wasn't used to it, this continual ability of the Perth Group to supply their own refutations would amaze me. Do they really want me to read the rest of them…?"
It is true that phytohemaglutinin (PHA) is a commonly used mitogen but it is also true that it is an oxidising agent. Furthermore, while the "control cultures" were stimulated only with PHA, the "HIV" infected cultures were stimulated not only with PHA, but also with other agents including "a feeder cell layer of 105 to 106 irradiated (4000 rads) PBL pooled from 10 to 20 normal human donors".
If we are going to have a scientific debate then Nicholas Bennett must read all the references in our rapid response: "Montagnier's reverse transcriptase activity", 23 July. Furthermore, the paper should not be just read but all the evidence critically analysed and faithfully reported.
Zagury et al wrote: "As shown in Fig. 1, from 0.5 to 3.0 per cent of the PHA-activated, RT-positive cultures of T cells from AIDS patients also expressed p15 and p24 HTLV-III antigens, identified by indirect immunofluorsecence with specific monoclonal antibodies."
In fig. 1 where the evidence for antigenic differences is presented, no controls, much less proper controls can be found.
Nowhere in the Zagury, Gallo et al 1986 paper, is there any evidence that the "Infected cells, once activated, died due to the effects of infection. Uninfected cells did not". To the contrary. By manipulating the culture conditions, "the HTLV-III-infected T cells could be maintained in the presence of exogenous IL-2 for 50 to 60 days….after this period, cell degeneration occurred in a manner similar to that of normal T cells grown under the same conditions. The cultures of infected T cells showed other similarities to cultures of normal T cells. When T-cell surface antigens were measured by the rosette technique in the presence of specific monoclonal antibodies, the proportion of infected cell cultures exhibiting T4 and T8 and Tac antigens were in the same range as normal T cells".
Although both Nicholas Bennett and Christopher Noble read the Zagury, Gallo et al paper and made a number of comments for some unknown reason, they did not mention a crucial experiment. Zagury et al stated: "Stimulation with PHA also induced variations in the T-cell differentiation antigens, such as loss of the T4 and Tac (IL-2 receptor), on day 6. This loss was more marked in the HTLV-III-infected cells (Table 3), although the differentiation of infected cells paralleled that of the normal uninfected cells."
Table 3. Antigenic variation in long-term cultured T cells from normal donors after infection with HTLV-III in vitro. The cultured (20 days) T cells were activated with PHA in the presence of macrophages and B cells and either uninfected or infected with HTLV-III as described in Table 2. Nonactivated normal T cells were also used as controls. T4 and Tac antigens were tested in triplicate for each culture on days 0, 2, and 6 after PHA stimulation. Variability in these samples was less than 10% and the results presented are the mean values.
*The High percentage of Tac+ (IL-2 receptor-positive) cells on day 0 of the
experiment is due to the previous long-term culture (20 days) of these cells.
As the table shows at day zero the number of T4 cells in all cultures was 34%. By day 6 the T4 cells in PHA stimulated non-infected cultures had decreased to 10% while in the PHA stimulated "infected" cultures the decrease was to 3%. Over the same time period the non-stimulated, non-infected cultures decreased to 25%. While the non-infected cultures were stimulated only with PHA, the "infected" cultures, in addition to PHA, were also treated with polybrene, supernatant fluids, IL-2, antibody to a-interferon, and hydrocortisone. This means that the difference in the T4 cell number between the PHA stimulated but non-infected cultures and the PHA stimulated and "infected" cultures may be due to the additional stimulation to which the latter were exposed.
In fact by 1983 Zagury himself has shown that following stimulation T4 cells may change to T8: "Thus, it appears that T antigens, which seemed to be molecular markers of differentiation, are not markers for terminal differentiation and do not always reflect defined functional properties" (1).
Zagury, Gallo et al also reported that "immunological activation is required for viral expression". That is, none of the phenomena which are said to prove "HIV" infection could be detected in the absence of stimulation (oxidation).
Since: (i) there is no decrease in T4 cells in "HIV infected" cultures unless stimulating agents are also added to the cultures; (ii) the stimulating agents in the absence of "HIV infection" cause decrease in T4 cells;
the only scientific conclusion a scientist can draw is that, the stimulating agents and not "HIV" is the cause of the decrease in the T4 cell number (immune deficiency). In other words "HIV" is neither necessary nor sufficient for the induction of immune deficiencies.
The last paragraph reads: "Our results also suggest that multiple rounds of antigenic stimulation in vivo, as a result of infection with various microorganisms or exposure to allogeneic cells such as semen or blood, may promote HTLV-III expression, T4 cell death, further spread of the virus, and ultimately an immunodeficiency syndrome."
Since: (i) "HIV infection" does not lead to decrease in T4 cell number unless stimulating agents are also present; (ii) the stimulating agents in the absence of "HIV" cause decrease in T4 cell number; (iii) AIDS patients are exposed to many stimulating agents;
there can only be one scientific conclusion: the ultimate cause of T4 decrease (immune deficiency, and thus AIDS) is not "HIV" but the many (semen, blood, micro-organisms, drugs) stimulating agents to which the patients belonging to the risk groups are exposed.
1. Zagury D, Bernard J, Morgan DA, Fouchard M, Feldman M. Phenotypic Diversity within Clones of Human Normal T Cells. Internat J Cancer 1983;31:705-710.
Competing interests: None declared