Infectious Disease Postdoc/Clinician
Department of Pediatrics, University Hospital, Syracuse NY
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I appreciate the Perth Group's response, which has helped clarify a number of points.
With reference to the Richieri Vaccine paper, I will find another copy (my original being left in the UK) and get a scanned image with comments sent as soon as possible. From memory however, the main image contains what I would describe as "wall-to-wall virions". From memory I do not recall the characteristic "spikes" of gp120, but since the vaccine prep in question was gp120-depleted I do not think this is relevant! The group did analyse the protein content and found "p6, p7[NC], pI15, p17[MA], p24[CA], p32, pI39Gag, gp41, pp55Gag, p66/51, Vpr, Vif and Nef" (square brackets added for clarity of protein identification).
There are many other suitable papers available, I suggest the Perth Group run a search for Larry Arthur, or consider the ref below ..
The main point is that using purification methods superior to simple density gradient centrifugation, they isolated pure particles which contained the viral elements. The secondary point is that they HAD to use superior methods, and that asking for HIV purification using sucrose density gradient purification is bound to fail!
Also, the fact that they were using gp120 depleted and UV-inactivated virions points to the fact that the field has moved beyond trying to isolate HIV.
The Gelderblom article makes no mention of using blood samples directly for analysis, but I do not doubt that if only 10^6 particles/ml maybe required then even after purification there might well be sufficient material left to analyse, after concentration. Reference 27 from that paper suggests that clinical isolates might well have been EM'd by Gelderblom himself, back in 1994, although not having attended the conference I cannot comment on whether the viruses were passaged prior to analysis . Perhaps it would be worth asking the man himself? I cannot possibly claim to be anything but a commentator on HIV EM's!
As regards Montagnier's RT data, this opens up a whole new thread of discussion. I refer back to a Perth Group paper published in the magazine Continuum in Sept/Oct 1996 where they state:
(a) the presence of reverse transcriptase (RT) is proven indirectly, that is, by demonstrating transcription of the RNA template-primer A(n).dT15;
(b) the template-primer A(n).dT15 can be transcribed not only by RT but by other cellular DNA polymerases. All the cellular DNA polymerases, alpha, and gamma, can copy A(n).dT15. In fact, in 1975, an International Conference on Eukaryotic DNA polymerases, which included Baltimore and Gallo defined DNA polymerase gamma, "a component of normal cells", "found to be widespread in occurrence", whose activity can be increased by many factors including PHA stimulation, as the enzyme which "copies A(n).dT15 with high efficiency but does not copy DNA well"; it is impossible to say that the polymerase in the "growth medium" or in the material banding at 1.16gm/ml which catalyses reverse transcription of A(n).dT15 is RT or one of a number of other cellular DNA polymerases.
This they use as evidence that Montagnier didn't adequately prove that RT had been detected. In fact, as happens frequently in Perth Group articles I find, the evidence to counter such conclusions is to be found in the very references they use to support their argument, refs 2-5 in the original article [2-5 below also], which I looked up some 5 years ago.
In summary: Montagnier's RT prefers Mg2+ at 5mM and pH 7.8
DNA pol gamma prefers Mn2+ and a pH of 7.4
The key word is PREFERS. DNA pol gamma will work with Mg2+ at pH 7.4, but Montagnier was careful enough to check they were the optimal conditions, and for DNA pol gamma they are not.
Additionally, DNA pol alpha does not transcribe poly A-Oligo dT, but early reports stated it did because of contamination with DNA pol gamma.
Montagnier also used further confirmatory methods, such as inhibition with Actinomycin D, which does not affect RT but does inhibit cellular DNA polymerases and RNA polymerases by binding to double-stranded DNA, which the Perth Group do not try to explain.
The age of the debate (which stretches right back to Duesberg's comments in 1983) is relevant because to most scientists the issues raised were questionable at the time, and have been addressed since. Refusing to accept the results of ongoing and more recent work does not equate to not having had the questions answered!
The other forums where these points were raised have been the sci.med.aids moderated newsgroup (until September 1993 - I am a past co- moderator of the group) the Re-Appraising AIDS and Re-Thinking AIDS email lists (of which I am a past and current subscriber respectively) and the misc.health.aids unmoderated newsgroup (Sept 1993 to date). MHA was established specifically to debate these issues and the Google archives contain tens of thousands of postings concerning AIDS controversies. The unmoderated group sci.med has also seen some relevant posts, but is not dedicated to AIDS in particular. Many of the references and facts I use in these rapid responses are in fact drawn from the archives of the MHA postings, highlighting the unending and circular nature of these discussions.
Despite repeated requests (so I'm told) the Perth Group has never to my knowledge posted anywhere to answer questions or confirm current opinions or refute criticisms of their work. The only name I recognise from the current Perth Group members who I have seen in debate is Todd Miller, from the Re-Appraising list and MHA, prior to when he co-wrote a paper with the other Perth Group members on the toxicity of AZT. I personally applaud the Perth Group for writing in these Rapid Responses, to enable a more fluid exchange of ideas and facts than was possible previously.
1. Gelderblom HR, Asjö B, Moore JP, Ho DD, Sandford J, Pauli G (1994). Are clinical isolates morphologically different from laboratory strains of HIV? 10th Interntational Conference on AIDS. 1994; Yokohama. AIDS, Abstract PA0195, Vol. 2, Yokohama.
2.Sarngadharan MG, Robert-Guroff M, Gallo RC. DNA polymerases of normal and neoplastic mammalian cells. Biochim Biophysica Acta 1978;516:419-487.
3.Weissbach A, Baltimore D, Bollum F, et al. Nomenclature of eukaryotic DNA polymerases. Science 1975;190:401-402.
4.Robert-Guroff M, Schrecker AW, Brinkman BJ, et al. DNA polymerase gamma of human lymphoblasts. Biochem 1977;16:2866-2873.
5.Lewis BJ, Abrell JW, Smith RG, et al. Human DNA polymerase III (R- DNA): Distinction from DNA polymerase I and reverse transcriptase. Science 1974;183:867-869.
. Ott et al. AIDS Research and Human Retroviruses Vol 11, Number 9, 1995. "Analysis and Localisation of Cyclophilin A found in the virions of Human Immunodeficiency Virus Type 1 MN Strain." pp1003-1006.
Competing interests: None declared