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Does HIV exists (fowarded from sci med aids) 2/3

weissm at rockvax.rockefeller.edu weissm at rockvax.rockefeller.edu
Thu Mar 9 16:33:43 EST 1995


Date:         Thu, 9 Mar 1995 14:05:46 -0600
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From: sma at wubios.wustl.edu
Subject:      Does HIV exist?
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To: Multiple recipients of list AIDS <AIDS%RUTVM1.BitNet at pucc.PRINCETON.EDU>

banded at 1.16 gm/ml, two, p41 and p24,  were found to react with some
patient sera. They concluded: "p24 and p41 may therefore be considered viral
structural proteins";
 4. Finally, they used the ELISA [Footnote 2] technique to test for HTLV-III
antibodies. 88% (43/49) of patients with AIDS, and 79% (11/14) patients with
pre-AIDS but "less than 1 percent of heterosexual subjects", had antibodies
"reactive against antigens of HTLV-III". "To understand the molecular nature
of the antigens recognized by ELISA", the sera were analysed by WB. "...the
antigen most prominently and commonly detected among all of the sera from
AIDS patients had a molecular weight of 41,000 (p41)...Reactivity to p24 of
the virus was generally very weak and was clear only in two cases". >From
the above data it is obvious that by HTLV-III (HIV) isolation was meant
detection of more than one of the following phenomena:
 1. RT, either in the culture fluids, or in the material from these fluids
or cellular lysates which in sucrose density gradients band at 1.16 gm/ml;
2. In culture fluids, but not in the material which bands at 1.16 gm/ml,
particles with morphological characteristics of retroviruses (RVP);
 3. Proteins, (p41, and in some cases, p24), which, in sucrose density
gradients, band at 1.16 gm/ml, (but without proof that they are unique
constituent parts of the particle), and react with patient sera.
 However, isolation is defined as separating an object, (HIV), from
everything else, and not the detection of some phenomena attributed to it
(RT, WB), or similar to it, (RVP). Phenomena can only be used for retroviral
detection, not isolation, and even then if, and only if, it is first shown
that each is specific for the virus by use of the only valid gold standard,
HIV itself, "HIV isolation". It is important to note that in the earlier
(1983) report by Montagnier's group on HIV (LAV) isolation, the same
experimental procedures and findings as those described by Gallo were
reported. The only exception was that Montagnier's group did not "infect" an
immortalised cell line, yet Gallo's group considered that Montagnier and his
colleagues had not described "true isolation".6 In fact, in 1984, evidence
existed that RT, antigen-antibody reactions (WB), and RVP, are non- specific
for retroviruses. The indirect evidence, that is, evidence that has been
obtained without a gold standard from recent AIDS research, has confirmed
the above.
Reverse transcriptase. Although Gallo has described the enzyme reverse
transcriptase as "unique to retroviruses", this is not the case, a fact
stressed by its discoverers, (both Nobel laureates).17 Reverse transcription
can be found in leukaemic T-cells,24 (HT and its clones including H9, from
which the first "HTLV-III (HIV) virus was isolated", is a leukaemic cell
line), normal spermatozoa,25 and, according to Harold Varmus, another Nobel
laureate, more recently, in the uninfected cells of yeasts insects and
mammals.26 As far back as 1973, Gallo himself was the first to show that RT
can be found in "PHA stimulated (but not unstimulated) normal human blood
lymphocytes".24  Confirmation of this was reported at the 1991 Florence AIDS
conference where evidence was presented that the drug AZT can inhibit the
action of normal cellular RT,27 and this was postulated as a mechanism for
drug toxicity.
 Retroviral particles. By definition, retroviral particles are enveloped
infectious particles 100-120nM in diameter with a core compromising a
protein shell and a ribonucleoprotein complex. RVP are further catergorised
according to the site of core assembly, that is, within the cytoplasm or at
the cell membrane, and by certain other morphological features. Included in
this taxonomy are the Subfamilies Oncoviruses which include Type C and Type
D particles, as well as the Subfamily Lentiviruses.
 Prior to the AIDS era, many retrovirologists showed that the finding of a
particle with morphological features similar to retroviruses does not
constitute sufficient proof that they are retroviruses, that they are
infectious particles, even if they are found to band at 1.16 gm/ml.18  In
1976 Gallo himself pointed out that in human leukemic tissue "virus-like
particles morphologically and biochemically resembling type-C virus but
apparently lacking the ability to replicate, have been frequently
observed".28  Particles with the morphological characteristics of
retroviruses were reported in milk, cultures of embryonic tissues and "in
the majority, if not all, human placentas".29,30,31  However, they were
considered to be "an intriguing and important problem that remains to be
solved".32  Evidence from AIDS research shows that:
1. There is no agreement on the precise taxonomic classification of HIV.
Initially, HIV was reported as an Oncoviral type-C particle,5 then a type-D
particle,33 and ultimately as a member of a different Subfamily, a
Lentivirus;34
 2. The T-cell and monocyte "HIV infected cultures" contain in addition to
particles with morphologies attributed to HIV, many other "viral particles"
unlike any of the "HIV particles".35,36,37,38  "Non-HIV-infected" HT (H9)
cells, the cell line from which the Gallo team "isolated" the first HIV
(HTLV-III) and from which most of the published electron micrographs of "HIV
particles" have originated, as well as other cells used for "HIV isolation",
CEM, C8166, EBV transformed  B-cells, and cord blood lymphocytes, express
virus-like particles albeit they are somewhat different from the variety of
particles accepted as HIV.39  The above data raises questions not only in
regard to the origin and role of the "non- HIV particles", but also to the
"HIV (HTLV-III) particles". Furthermore, neither Gallo's team, nor anybody
else before or since has published EM micrographs of the material derived
from AIDS cultures/co-cultures which bands at 1.16 gm/ml. Thus it is
impossible to know which, if any of the particles, band at that density;
3. Most importantly, it is generally accepted that particles reported in the
lymph nodes of AIDS patients are HIV. However, in the only EM study40,
either in vivo or in vitro, in which suitable controls were used and in
which extensive blind examination of controls and test material was
performed, "HIV particles" were found in 90% (18/20) of patients with
persistent generalised lymphadenopathy attributed to HIV, and in 87% (13/15)
of patients with "non-HIV lymphadenopathies", leading the authors to
conclude: "The presence of such particles do not, by themselves indicate
infection with HIV". Antigen-antibody reactions. One can claim that a given
protein is an antigen derived from an exogenous retrovirus if first it is
shown that:
1. The protein is a structural component of a particle;
2. The particle is a retrovirus;
3. The protein is coded exclusively by a viral and not a cellular gene.
 Once the above are demonstrated, the only way to prove that the antibodies
found in AIDS patient sera are directed against the viral antigen is to use
the antigen or the isolated virus as a gold standard. The mere finding that
a protein from the AIDS cultures bands at 1.16 gm/ml and reacts with sera
from AIDS patients cannot be considered to simultaneously prove that:
 1. The protein is a viral antigen;
 2. The antibodies in the AIDS patient sera which react with the antigen are
specific for that antigen.
At present, it is known that about 80% of the proteins which band at 1.16
gm/ml, some of which react with some AIDS sera, do not constitute any of the
proteins ascribed to HIV.41,42,43 Most importantly, prior to the publication
of the Science papers, evidence existed, confirmed since, which is at odds
with the conclusion that "p24 and p41 may therefore be considered viral
structural proteins":
 The p41/45 protein In AIDS research, the p41 and p45 bands are considered
to represent one and the same HIV protein.
1. Like Gallo's group, Montagnier's team one year earlier, found that AIDS
sera reacted with a protein p41/45 from the AIDS cultures and which in
sucrose density gradients, banded at 1.16 gm/ml. However, from their data
they considered that the p41 band "may be due to contamination of the virus
by cellular actin which was present in immunoprecipitates of all cell
extracts",5 that is, of "HIV infected" as well as non-infected cells and
cells infected with HTLV-I. Although Gallo's group did not find such a
reaction with p41 in non-infected cells, they did find a p80 protein and
concluded that the reaction was "non-specific".8  However, at present it is
known that p80 as well as two additional "HIV proteins", p120 and p160, are
oligomers of p41.44  Which protein (band), p41, p80, p120 or p160 is
detected in a given WB depends on the culture and WB conditions, including
temperature and the concentration of sodium dodecyl sulphate used to disrupt
the proteins which band at 1.16 gm/ml;45
2. Actin is an ubiquitous protein present in all cells including bacteria
and several viruses. Well known retroviruses such as the mouse mammary
tumour virus have also been shown to contain actin of cellular origin and it
has been postulated that this protein plays a key role in both retroviral
assembly and budding;46,47
3. Platelets from healthy individuals also contain a p41 protein which
reacts with sera from homosexual men with AIDS and immune thrombocytopenic
purpura (ITP) and which "represents non-specific binding of IgG to actin in
the platelet preparation".48
 4. Researchers at the Pasteur Institute have shown that sera from AIDS
patients and AIDS risk groups contain high levels of antibody against calf
striated muscle actin.49
The p24/25 protein 1. Apart from a joint publication with Montagnier where
they claim that the HIV p24/25 is unique, Gallo and his colleagues have
repeatedly stated that the p24s of HTLV-I and HIV immunologically
cross-react;50
2. Genesca et al51 conducted WB assays in 100 ELISA negative samples of
healthy blood donors; 20 were found to have HIV bands which did not fulfil
the then (1989) criteria used by the blood banks for a positive WB. These
were considered as indeterminate WB, (WBI), with p24 being the predominant
band, (70% of cases). Among the recipients of WBI blood, 36% were WBI 6
months after transfusion, but so were 42% of individuals who received
WB-negative samples. Both donors and recipients of blood remained healthy.
They concluded that WBI patterns "are exceedingly common in randomly
selected donors and recipients and such patterns do not correlate with the
presence of HIV-1 or the transmission of HIV-1", "most such reactions
represent false- positive results";
 3. Antibodies to p24 have been detected in 1 out of 150 healthy
individuals, 13% of randomly selected otherwise healthy patients with
generalised warts, 24% of patients with cutaneous T-cell lymphoma and
prodrome and 41% of patients with multiple sclerosis;52
4. Ninety seven percent of sera from homosexuals with ITP and 94% of sera
from homosexuals with lymphandenopathy or AIDS contain an antibody that
reacts with a 25Kd membrane antigen found in platelets from healthy donors
and AIDS patients, as well as a 25 Kd antigen found in green-monkey kidney
cells, human skin fibroblasts, and herpes simplex cultured in monkey kidney
cells. This reaction was absent in sera obtained from non-homosexual
patients with ITP or non-immune thrombocytopenic purpura;48
5. Conversely, the p24 antigen is not found in all HIV positive or even AIDS
patients. In one study, the polymerase chain reaction (PCR) and p24 were
used to detect HIV in patients at various CDC stages from asymptomatic to
AIDS. p24 was detected in 24% patients and HIV RNA in 50%;53
6. In another study, "In  half of the  cases in which a subject had a
positive p24 test, the subject later had a negative test without taking any
medications that would be expected to affect p24 antigen levels...the test
is clinically erratic and should be interpreted very cautiously".54 Thus the
finding of viral particles in the AIDS cultures/co- cultures, RT and
proteins which react with AIDS related sera in the material from the
supernatant or cell lysates which in sucrose density gradients bands at 1.16
gm/ml, cannot be considered synonymous with the isolation or even the
detection of a retrovirus. Even if a retrovirus is isolated from in vitro
cultures/co-cultures from tissues from AIDS patients, this does not, by
itself, constitute proof of the existence of the virus in vivo, (in AIDS
patients), and even less that the retrovirus has been exogenously acquired.
This is because:
1. At present, it is generally accepted that "one of the most striking
features that distinguish retroviruses from all other animal retroviruses is
the presence, in the chromosomes of normal uninfected cells, of genomes
[proviruses] closely related to, or identical with those of infectious
viruses". The human genome, in addition to other proviral sequences, is
known to contain both HTLV-I55,56 and HIV57 sequences. Depending on
conditions, the proviral genome remains unexpressed or part or all of it may
be expressed. The latter may or may not lead to the assembly of viral
particles (endogenous retrovirus).17  In animal cultures, healthy non-virus
producing cells sooner or later spontaneously release retroviruses.20  The
appearance and yield can be increased by (i) mitogenic stimulation;58 (ii)
co-cultivation techniques;59 (iii) cultivation of cells with supernatant
from non-virus producing cultures.60  According to one eminent
retrovirologist, George Todaro, "the failure to isolate endogenous viruses
from certain species may reflect thelimitation of in vitro cocultivation
techniques";61
2. Gallo's team, like everybody else: (i) "isolated HTLV-III (HIV)" from
cell cultures; (ii) "isolated HTLV-III" from mitogenically stimulated,
activated cell cultures;
3. In addition, Gallo and his colleagues also used co-cultivation
techniques;
4. The first "HTLV-III isolation" was from the HT (H4, H9, H17) cell line.
Reading Gallo and his colleagues' first paper, one surmises that the HT cell
line was established in Gallo's laboratory. The Gallo inquiry revealed that
the HT cell line is in fact HUT78, a cell line established in another
laboratory from a patient with mature T4-cell leukaemia, a disease which
Gallo claims is caused by the exogenous retrovirus, HTLV-I.3  If so, then
all HT cell cultures, and the clones derived from it, "infected with
HTLV-III" or non-infected, and the material from these cultures which bands
at 1.16 gm/ml, should contain HTLV-I, and thus RT and retroviral particles.
Furthermore, because about 25% of AIDS patients have antibodies to HTVL-I,1
and the immunogenic proteins of HTLV-I and HIV have the same molecular
weights, then approximately 25% of the non-infected HT (H4, H9, H17)
cultures in addition to RT and particles, should have, in the Western blot,
the same bands as those of the "HTLV-III infected" cultures. Thus, these WBs
will erroneously appear positive for HTLV-III.
 Proof that HTLV-III (HIV) is causally linked to AIDS. Gallo claims, a claim
accepted by the vast majority of AIDS researchers, that in the May 1984
Science papers he and his colleagues presented "unambiguous evidence that
this [virus] and this alone was the cause of AIDS".62  A minimum requirement
for making such a claim should be presentation of the following evidence:
1. That all AIDS patients are infected with HTLV-III;
2. Infection with HTLV-III leads to T4-cell depletion, given the assumption
that HTLV-III leads to the clinical syndrome by its T4 cytotoxicity.
 The evidence for the existence of HTLV-III was "viral isolation" and ELISA
antibody tests. Even if one assumes that the data presented represents "true
isolation", the virus was isolated from less that half (10/21) of AIDS
patients with opportunistic infections, and in less than one third (13/43)
with Kaposi's sarcoma, then and now the two most characteristic AIDS



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