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
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IS OXIDATION NECESSARY FOR ACTIVATION?
In our rapid response, "Montagnier's reverse transcriptase activity", 23rd July, we wrote: "Let's assume that using the template-primer poly(a).dT15, "ph 7.8" and "5mM MgCl2", Nicholas Bennett [like Montagnier before him] detects reverse transcriptase activity in: (i) cell cultures; (ii) a "purified" virus preparation which actually contains cellular vesicles (cellular fragments) but in which no retroviral particles can be seen even after Roman effort;
What scientific reasons would he have to conclude that the reverse transcriptase activity detected in the cultures or in the "purified" virus, is due to the enzyme reverse transcriptase of a retrovirus and not to the cellular DNA polymerases?"
Nicholas Bennett responded in his rapid response. "Re: Montagnier's reverse transcriptase activity", 26th July but did not answer our question, so, we repeat our request to Nicholas Bennett.
In the same rapid response we wrote: Would Nicholas Bennett please read references 2-7 and tell us if: (i) to stimulate cells and to obtain permanent cell lines, is or is not oxidation necessary; (ii) Montagnier, in 1983 and Gallo in 1984, treated their cells (Bru's lymphocytes, the umbilical cord blood lymphocytes and the H9 (HUT-78) cell line) with oxidising agents; (iii) Montagnier and Gallo consider stimulation (oxidation) necessary for the "production of viral ["HIV"] particles, antigenic expression and the cytopathic effect".
(1) Rey, M.A., Spire, B., Dormont, D., Barre-Sinoussi, F., Montagnier, L., Chermann, J.C. (1984), Characterisation of the RNA dependent DNA polymerase of a new human T lymphotropic retrovirus (Lymphadenopathy Associated Virus). Biochemical & Biophysical Research Communications, 121,(1), 126-133.
(2) Sekkat, C., Dornand, J & Gerber, M. (1988), Oxidative phenomena are implicated in human T-cell stimulation. Immunology, 63, 431-437.
(3) Papadopulos-Eleopulos, E. (1982), A mitotic theory. J. theor. Biol., 96, 741-758.
(4) Barré-Sinoussi F, Chermann JC, Rey F, Nugeyre MT, Chamaret S, Gruest J, et al. Isolation of a T-lymphotropic retrovirus from a patient at risk for acquired immune deficiency syndrome (AIDS). Science 1983;220:868-71.
(5) Popovic M, Sarngadharan MG, Read E, Gallo RC. (1984). Detection, Isolation and Continuous Production of Cytopathic Retroviruses (HTLV-III) from Patients with AIDS and Pre-AIDS. Science 224: 497-500.
(6) 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.
(7) Klatzmann, D & Montagnier, L. (1986), Approaches to AIDS therapy. Immunology, 319, 10-11."
Nicholas Bennett responded: "I am sure that stimulation of primary cells is necessary, but I take issue with the Perth Group's persistence in saying "stimulation (oxidation)". The two terms are NOT synonymous, and linking them in this way is extremely misleading. Many mitogens act not through "oxidation" directly but through specific receptors and cellular signalling pathways. In fact, T cell responses and IL-2 production are reduced by oxidative reagents  despite oxidative agents increasing the effects of NF-kB" As well as showing how a sledgehammer approach can affect multiple pathways simultaneously, it suggests that an activating cytokine (one kind of mitogen) like IL-2 is far more subtle."
We never said that oxidation is synonymous with stimulation. We claim that for stimulation oxidation is absolutely necessary. However, oxidation does not necessarily lead to stimulation. One can invent as many "receptors" and "signalling pathways" as he/she desires. The fact is that stimulation requires oxidation (refs. 2 & 3). It is also a fact that when the oxidation is too "harsh", instead of the cell being stimulated they will die and IL-2 production will cease (ref. 3).
Nicholas Bennett wrote: "As such without reading the references provided I know from my education that it's obvious that a great many cell lines DO require stimulation in order to become established, and agree with the Perth Group on this. In this situation, some of the agents used are quite harsh, and I'm prepared to accept the use of the word "oxidising" in this context for certain cell lines, although I can't comment on the specifics of H9 and the parent HUT-78."
We are glad that there is some agreement between us.
Nicholas Bennett wrote: "However, stimulation of cells to activate HIV is an entirely different situation, and shouldn't be included haphazardly."
Would Nicholas Bennett please read ref. 6 and 7 and tell us if stimulation is or is not necessary to activate "HIV".
Competing interests: None declared