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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.”
===
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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