Reprinted from Continuum Magazine, Winter 1999
Virtual Viral Load Tests: Seeing is Believing
By Michael Verney-Elliott
 
“HIV, if detectable at all, is only ever found in minute trace quantities, and even then only by stretching laboratory culture techniques to their limit.”
 
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As Peter Duesberg pointed out in his ground-breaking paper "Retroviruses as Pathogens and Carcinogens: Expectations and Reality"(Cancer Research, 1.3.87), one of many reasons why HIV could not be the cause of AIDS is that the virus is never found in sufficient quantity to have any pathogenic significance in those supposedly infected.
 
Typically, viruses that cause disease are found at very high titre (concentration or strength) at the time the disease is active, but this just did not seem to be the case with HIV, either in those people said to be incubating AIDS for ten years or those with the full-blown condition. By contrast, supposed HIV, if detectable at all, is only ever found in minute trace quantities, and even then only by stretching laboratory culture techniques to their limit. Minute quantities is one of the chief characteristics of a persistent, harmless passenger virus.
 
The currently accepted evidence for the presence of active HIV depends on surrogate markers, signals, non-specific antibodies etc., and what Jon Cohen calls "...HIV RNA's – a proxy for the amount of free virus ...".(Science, 13.1.95, p.179) Thus the HIV viral titres are deemed to be present by inference rather than direct observation as is usually the case with other disease causing microbes. As Lady Bracknell might have said: "A proxy? Viruses should be seen, not inferred!"
 
In 1995, David Ho and Xiping Wei published separate papers in the same issue of Nature (373, pp. 123 et seq. and 117 et seq.) claiming that far from being the indolent virus supposed during the previous 11 years of AIDS research, HIV was hyperactive, and soon after infection, high titres of virus were circulating continuously in the peripheral blood of HIV positive individuals. Although these high titres had never been seen before by any other AIDS researchers, by amplifying “viral RNA” using PCR and using the resulting DNAs as a proxy, the two papers claimed that the corresponding viral RNA's represented the amount of cell-free virus present in the blood. No one spotted the absurdity that if there were that much virus present in the blood you would not need to amplify it by PCR in order to detect it.
 
However, the scientific community uncritically accepted this purely hypothetical theory of “HIV dynamics” as conclusive evidence for it being the cause of AIDS, to the extent of making these papers the basis for the so-called viral load tests currently used, and the prescription of “anti-viral drugs” like protease inhibitors and AZT. Both papers have been thoroughly debunked by Duesberg and Bialy (Nature, 375, 1995, p.197), by Paul Philpott and Christine Johnson (Reappraising AIDS, Vol.4, No.10, October 1996), and more recently by Roederer et al. (Nature Med 4, 145,  1997), and Hellerstein et al. (Nature Med. 5, 33, 1999).
 
Professor Etienne de Harven, a specialist in electronmicroscopy, has written two articles for Continuum (Vol. 5, No 2, Winter 97; Vol. 5, No.3, Spring 98) describing the standard method he developed in the 1960's for visual detection and identification in fresh plasma of “RNA tumour viruses,” the name by which retroviruses were once known. The procedure can be carried out quite simply in any lab equipped with facilities for centrifugation, microfiltration andelectron microscopy (EM) as follows:
 
- A blood sample suspected of containing a pathogenically significant quantity of cell-free retroviruses is centrifuged to spin out the red and white blood cells, leaving just the plasma, the basic fluid in which the cells float.
 
- The fresh plasma is then diluted 1/1 with cold heparinised Ringer's solution. Heparin is an anticoagulant, added to thin the plasma and render it more suitable to pass successively through two millipore filters of diminishing size, first using 0.6 then 0.22 micrometer pore size diameter filters. These filter out any debris larger than the size of the viral particles being sought. The last filter used will be slightly larger than the 100-120 nm diameter of a retrovirus.
 
- The final filtrate is then spun in a centrifuge at 30,000 g for two hours. This has the effect of concentrating any virus in the plasma into a tiny pellet that can be recovered from the bottom of the test tube.
- The pellet is then fixed, embedded in an epoxy resin block and shaved into ultra thin slices which can be mounted on copper grids and examined under EM.
 
A perfect illustration of de Harven's isolation technique is seen in an electron micrograph published in 1965, (Pathologie-Biologie, Vol.13, pp. 125-134) and shows densely packed retroviral particles of the Friend murine (mouse) leukaemia virus. The densely packed particles, identical in shape and size, were pelleted down from fresh plasma using the method described. The magnification is 19,500 x and it can be seen that there is very little extraneous material contaminating the final pelleted isolate, indicated by three arrows. The Friend virus is classified morphologically as a Type C oncovirus, one of a small group of retroviruses which encode a cancer-causing gene in their RNA.  The ultra thin slice through the pellet clearly shows the dense core in bisected virions as round black dots.
 
As Dr. de Harven explains in his second Continuum article, even by the late 1960s there was a growing tendency to abandon the use of EM on the pretext that it was cumbersome and time-consuming, in favor of the purification and identification of viruses using other means, chiefly the use of the sucrose density gradient, allied with indirect surrogate markers. Already, the virologists were starting to cut corners.
 
Moreover, after 1970, when the race was on to find human retroviruses which cause cancer, there was a very good reason why clear, EM visualisation of such human oncoviruses was abandoned.  The simple truth was that they were unable, then as now, to find high titres of cell-free retroviruses in fresh human blood or plasma.
 
It is worth stressing at this stage that in the entire 15 years of HIV/AIDS research, no micrograph has ever been published purporting to show purified, densely packed HIV particles recovered from the fresh plasma of HIV positive subjects.
 
Indeed, no such picture exists of any so-called human retrovirus, not even HTLV 1, the alleged cause of adult T-cell leukemia. That is not to say that micrographs of alleged HIV have not been published, but they invariably came from cell cultures grown sometimes for weeks, in the total absence of an immune system (it is worth remembering that HIV infection is only diagnosed on the detection of antibodies supposedly specific to HIV), and involving co-culturing with known cancerous cell-lines such as H9 or CEM, and stimulation with mitogens, hydrocortisone and other chemical activators - the standard laboratory methods of reactivating latent viruses.
 
There is no better way to show the degeneration into sloppy imprecision in science characterized by the AIDS War than to compare de Harven's micrograph with a micrograph published in 1997 of material banded at 1.16.gm/ml in a sucrose density gradient.
 
Prior to its publication by Gluschankof et al. (Virology, 230, pp 125-133, 1997), it was claimed that material banding at this level represented pure retrovirus. However, this study, and another in the same issue by Bess et al., finally came clean and admitted that all sorts of debris and extraneous matter banded at the same level as retroviruses in the sucrose medium, principally cellular microvescicles, something that de Harven had observed even in the 1960s.
 
Note that whereas de Harven has to use three arrows to identify that which may or may not be retroviruses, Gluschankof et al. by total contrast have to use three arrows to identify three 'viral' dots in their culture-derived rubbish tip. The comparison strikingly illustrates the precision and superiority of de Harven's method of virus isolation and identification to currently used methods.
 
Undoubtedly, sedimentation in sucrose density gradients has its uses, particularly in confirming viral identity. Viruses have specific buoyant densities, and it is indeed known that purified retroviruses form a distinct sedimented band at 1.16 gm/ml, but it is a major error to suppose that all material contained in the gross supernatant from a cell culture which bands at that level is pure retrovirus.
 
Moreover, in plasma samples containing more than one variety of active virus, as in AIDS patients with several, concurrently active viral infections, distinguishing them by their morphology under EM may be difficult, so subsequent buoyant density tests may be needed. Even the use of surrogate markers may be legitimate, but only when numerous laboratories have established that a large quantity of virus particles, of identical shape and size, are invariably present in diseased tissues and the virus has been proved beyond any doubt to be the cause of the disease. Then, and only then, can markers or proxies be trusted to indicate viremia. However, 'HIV' fails all these tests.
 
Logically, if Ho and Wei are correct, it must be possible to pellet down fresh plasma from a person or persons who have had a recent “high viral load” test result and clearly see tightly packed identical viral particles using EM, in a quantity consistent with the amount of virus indicated by the so-called viral load test. The application of the de Harven methodology will clearly demonstrate the presence or absence of actual rather than virtual viral particles, and do away with bogus mathematical models, inappropriate use of PCR, proxies, surrogates and all the other trappings of modern scientific obfuscation. If the virus particles cannot be seen, then they are just not there, whatever the test may claim. If it is claimed that there are indeed HIV particles in the plasma, but too few to form a pellet, then there is obviously not enough virus in the sample to be pathogenically significant.
 
Had AIDS researchers been able to photograph high titres of cell-free HIV in fresh plasma using the de Harven method during the last fifteen years, undoubtedly they would have done so and published their results, even if only to silence critics like Peter Duesberg. There are no such micrographs in the entire AIDS literature.
 
When assessing how much virus - viable, enveloped, infectious virion units - should be visible from the amount of "HIV RNA's" determined by a currently used viral load test, important scientific evidence shows that the proxy viral RNA's represent very few actual potentially infectious viral particles. As Duesberg and Bialy state in the heavily censured version of their viral load theory critique published in Nature (Ibid): "The senior researcher [George Shaw] of the Wei et al. paper has previously claimed that the PCR method they used overestimates by at least 60,000 times the real titre of infectious HIV [Piatak et al., Science, 259, pp 1749-1754, 1993]. 100,000/60,000 is 1.7 infectious HIV's per ml... Further, Ho and a different group of collaborators have just shown [Cao, Y. et al. New Eng. J. Med. 332, pp 201-208, 1995] that more than 10,000 plasma virions, detected by the branched-DNA amplification assay used in their Nature paper, correspond to less than one (!) infectious virus per ml. And infectious units, after all, are the only clinically relevant criteria for a viral pathogen."
 
Modern microbiologists and virologists have developed, and continue to develop, a bewildering array of techniques to aid them in pursuit of their disciplines. However, the increasing sophistication of the technology carries with it a proportionally increased need for scrutiny and analysis of their lab results. Modern culture techniques, involving mitogenic stimulation, other chemical additives, co-culturing with known cancerous cell-lines, the use of sucrose density gradients - all these valuable modern tools of science can easily produce results open to misinterpretation, accidental or deliberate.
 
Add to this the pressure from financial interests typified by pharmaceutical companies seeking quick results to order, and pressure on labs to secure grant funding etc. and it is not difficult to see how a few virus-like particles, inevitably dredged up in cell cultures, can be parlayed into a massive viremia by using proxies and mathematical prestidigitation.
 
The uncritical acceptance of the viral load theory has led to some patently laughable studies. The typical absurdity came from Farzadegan et al. in The Lancet (7.11.98), who carried out a long-term study based on blood samples taken from 650 male and female injecting drug users (IDU's), principally African Americans, using three different methods of measuring viral load. The results of their trial claim to show that the women progress to AIDS at the same rate as men but with only half the amount of viral load. In other words, HIV is supposedly twice as pathogenic in females as males - yet another pathogenic first for HIV. At no stage do they mention the possibility that the harmful effects of the continuous use of illicit drugs far more logically explains the seeming equality of progression to AIDS in both sexes, irrespective of apparent differences in HIV kinetics between the sexes based on ambiguous viral load tests.
 
Unless their data are confirmed by similar studies of HIV positive African, Asian and European males and females who are not IDU's, what does their study prove? Chillingly, the paper suggests that as the virus appears to be doubly pathogenic in women, they should be urgently considered for early drug cocktail therapy as soon as diagnosed.
 
In the final analysis, the only way to establish a true, in vivo viral titre in peripheral blood is by recovery of virus from a measured quantity of fresh, suspect plasma, and seeing the packed particles in a micrograph. Seeing, in this instance at least, is believing. As de Harven has explained (Ibid), an aliquot of the unfixed viral pellet may be resuspended in Ringer's solution and used for titration by the precise, traditional method. Virus counting under EM may be tedious, but would, of course, reinforce the observations made. If few or no viral particles can be seen in the above conditions, then certain questions must be asked: If not whole infectious viral particles, what is the viral load test measuring?
 
What are the “proxy RNA's” representing? After administration of protease inhibitor drugs, and alleged decrease in proxy RNA's, what has in fact been inhibited? If little or no infectious virus is found in plasma of supposed viremic people, how did the haemophiliacs become infected by their plasma-derived clotting factors?
 
Thus it may be seen that the viral load test at best translates into a barely, if at all, detectable level of viral particles which can have little relevance in AIDS pathogenicity; at worst it is a specious argument to pump asymptomatic people full of expensive drugs which may cause more harm than good in the long run.

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