The Perth Group has been roundly criticised by Professor Weiss over the issue of purification to prove the existence of HIV. When will the Perth Group get over this obsession?
It is true that in his 1999 email correspondence Professor Weiss expressed the opinion that access to purified materials is not necessary to prove the existence of a retrovirus. In fact Professor Weiss wrote: "...I find the issue of 'purification' quite sterile, and unconnected with matters of medical importance [1]. We are simply talking at cross-purposes. There appears to be a consortium of medical people and biophysicists in Perth who have a fixation of HIV purification. Perhaps you are influenced by this. It is also a 'cause' championed by the British based magazine 'Continuum'..."
We respect the views of Professor Weiss but perhaps our diametrically opposed views on this particular matter simply reflect the fact we are not professional virologists. Which means that as non-experts either we accept the views of the established experts because of the authority invested in them or, if unconvinced, explore other avenues to obtain a satisfactory resolution. At present we stick to our "obsession" because:
As we explained in the previous question, unlike all "other viruses" retroviruses or retrovirus phenomena may arise de novo. In fact in 1976 the eminent retrovirologist George Todaro wrote: "the failure to isolate endogenous viruses from certain species may reflect the limitations of in-vitro cocultivation techniques" [2]. (Endogenously produced retroviruses are morphologically and biochemically indistinguishable from exogenous retroviruses). In other words, Todaro is hypothesising that if only a scientist knew how to manipulate his cell culture conditions he could make retroviruses appear in ANY cell culture. We can even imagine a Todaro prediction which forecast "In 1983 cell cultures conditions pertaining to certain groups of individuals (now known as the AIDS risk groups and whose cell cultures are manipulated in particular ways including ways that did not exist in 1976), will produce a retrovirus" [3]. Thus, given that particular chemistries (in patients and/or cultures) may induce the appearance of retroviruses, experiments must be performed to distinguish between (a) a retrovirus causing illness and the accompanying laboratory abnormalities; (b) the diseases or culture conditions causing the "retrovirus". To date, in relation to "HIV", there are no such experiments.
Since the chemical makeup (known or unknown) may induce the appearance of retroviruses the only way to prove that a particle which looks like a retrovirus is truly a retrovirus is to introduce pure particles into a fresh culture and prove identical particles are the result. Thus this is one reason why purification is necessary. Twice in fact. Once to procure pure particles and a second time to compare the new with the old. This experiment must also include rigorous controls in which cultures are obtained from patients similar to AIDS patients (but without AIDS) and are manipulated in exactly the same manner as the AIDS patient cultures. The failure to employ such controls is an unfortunate feature of all HIV/AIDS research.
If purification is not important for proving the existence of a new retrovirus then why did Montagnier and Gallo undertake a procedure (banding in density gradients see Endnote 4) which they claimed resulted in "purified virus"? [4]
Virtually all EMs of "HIV" published to date are of unpurified cell cultures. An example is here. This is the EM published by Gallo in his second Science paperin May 1984. Evidence published for the first and as far as we know only time in March 1997, shows that the material designated "HIV" has not been purified. Yet this is the material used to obtain the "HIV" proteins (for the antibody tests) and the "HIV" genome (RNA and DNA for the "viral load" and "viral burden" tests). See here [5,6]. In fact not only is it impure but the particles in this material claimed to be a retrovirus HIV do not possess all the morphological characteristics required by the Gelderblom definition. The predominant constituent of this "purified virus" material is "microvesicles", that is, cellular debris. Thus, even if the particles these scientists designate "HIV" are a retrovirus HIV it is impossible to obtain their proteins and nucleic acids (RNA, DNA) without first separating the particles from everything else, that is, purifying the particles. To revisit Shakespeare, Juliet cannot send her nurse into the garden to obtain the proteins or DNA or a rose if she cannot (a) identify a rose or does not even know if roses exist; (b) separate roses from all the other flowers in the garden. Yet the HIV experts claim to have identified both HIV and its biochemical constituents under the same circumstances.
Purification is necessary to act as a gold standard for the antibody tests. See a subsequent question.
We leave it up to the reader to decide whether or not purification is a matter of "medical importance".
Also read our unpublished commentary on Gallo RC, Montagnier L. The discovery of HIV as the cause of AIDS. New England Journal of Medicine 2003;349:2283-5.
ENDNOTES
Although Professor Weiss believes purification is unimportant other retrovirologists do not agree.
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, editors. Animal Virology. New York: Academic Press Inc.; 1976. p. 369-384.
The laboratory phenomena (retroviral-like particles, reverse transcriptase activity, antibody reactions, RNAs) which are detected in cell cultures/cocultures of tissues of AIDS patients have several explanations apart a novel (new), exogenously acquired retrovirus. Endogenous retrovirus is one such possibility but there are others. For a more detailed discussion see our response to Peter Duesberg here. Of interest is the fact that in 1994, writing in Harrison's Principles of Internal Medicine, Gallo and Fauci stated "...there are no known human endogenous retroviruses". One can only speculate what may have been the course of the AIDS era had these scientists at least considered the possibility that the laboratory phenomena they detected were non-specific. Possibly including what George Todaro was predicting. After all, cells are cells and cultures are cultures and if endogenous retrovirus can arise de novo or be coaxed out of animal cell cultures which are not infected from without then why not humans as well? Nowadays, in fact dating from 1993, scientists accept that at least 1% of the human genome is made up of endogenous retroviruses [7,8]. Note added February 2004: "Sequences of retroviral origin occupy approximately 8% of the human genome". See here.
In his 1983 Science paper Montagnier claimed that the lymph nodes of BRU contained HIV which he purified using a method known as density gradient centrifugation. In this procedure an aliquot of culture supernatant is layered on top of a column of sucrose solution prepared such that its density gradually increases from top to bottom. The tube is spun at very high speeds for several hours and if the supernatant contains retrovirus particles they aggregate (band) in that portion of the gradient where the density reaches 1.16 gm/ml. Montagnier and Gallo both reported their 1.16 gm/ml banded material as “purified” virus, that is, material which one would expect to consist of retroviral particles and little or nothing else. However, neither group published an EM of the banded "purified virus". (Such EMs of "other viruses" have been published. See Rous sarcoma virus here). In 1997 Montagnier admitted he had not published an EM of his "purified virus" because, despite a "Roman effort", he and his colleagues could not find any particles in the banded material "with the morphology typical of retroviruses". (Read the Djamel Tahi interview here). In other words, there were no retrovirus particles in Montagnier's "purified virus", a fact little known to the general public and not acknowledged by the CDC, NIAID or any other HIV expert. Gallo has never addressed the issue of why he did not publish an EM of his "purified virus" or whether he even took an EM. Therefore Gallo's evidence for the existence of retroviral-like particles in "purified virus" did not exceed that of his French colleague's.
Gluschankof P, Mondor I, Gelderblom HR, Sattentau QJ. Cell membrane vesicles are a major contaminant of gradient-enriched human immunodeficiency virus type-1 preparations. Virology 1997;230:125-133.
Bess JW, Gorelick RJ, Bosche WJ, Henderson LE, Arthur LO. Microvesicles are a source of contaminating cellular proteins found in purified HIV-1 preparations. Virology 1997;230:134-144.
Lower R, Boller K, Hasenmaier B, Korbmacher C, Muller-Lantzsch N, Lower J, et al. Identification of human endogenous retroviruses with complex mRNA expression and particle formation. Proceedings of the National Academy of Sciences of the United States of America 1993;90:4480-4484.
Lower R, Lower J. Endogenous retrovirus sequences in human teratocarcinoma cell lines. Journal of Acquired Immune Deficiency Syndromes 1993;6(No 6):738.
