| PaulKing 2005-03-29, 6:22 pm |
| Reinherz et al., 1980; Reinherz et al., 1981).
The conclusion in (a) and (b) are at odds with evidence published in the
1980s. In 1989 it was shown that when "monocytes adhered to plastic (but
not when cultured on Teflon), a significant decrease in CD4 expression was
observed between 1 and 24 h post-adherence. CD4 expression could not be
detected in macrophages adhered to plastic for 5 days by using four
anti-CD4 monoclonal antibodies in flow cytometry or direct
immunofluoresence. Conversely, an increasing proportion of adherent cells
expressed LeuM3 and OKM5 surface antigens over the 5 days". It was also
shown that:
(a) "The down-regulation of CD4 was post-translational";
(b) unlike monocytes cultured on Teflon, the adherence of monocytes to
plastic resulted in superoxide anion generation, that is, oxidative stress
(Kazazi et al., 1989).
In the early 1980s, many researchers found that under certain conditions,
while the number of T4 cells decreases, the number of T8 cells increases
and the total number of cells remains constant or even increases. In 1982
Birch et al showed that incubation of T lymphocytes with adenosine or
impromidine, (an H2 histamine agonist), leads to a decrease in the number
of T- cells expressing the CD4 antigen and to an increase in the number of
T-cell expressing the CD8 antigen whilst the sum (T4 + T8) remains
constant (Birch et al., 1982). In an experiment conducted in the same year
by Burns et al (Burns et al., 1982), normal human peripheral blood
lymphocytes from different subjects were grown in conditioned medium
containing IL-2, and, after varying periods of time in culture, the cells
were tested by indirect immunofluorescence for OKT4 and OKT8. The
"conditioned medium" (CM) consisted of "cell-free supernatant passed
through a bacterial filter" from 7-day cultures of PHA stimulated
leucocytes obtained from patients with hemochromatosis. "For some
experiments CM was freed of residual PHA by passage over a
thyroglobulin-Sepharose column". They found that "...the cell population
progressively increased in size to large blasts...but most striking was
the rapid change in the OKT4:OKT8 ratio of cells within the population,
from 60:40 to 40:60...The change in the surface phenotype of the major
population also occurred in cultures maintained in medium containing IL2
which had been freed of PHA". They also found that the "change in
phenotype of the culture as a whole took place very rapidly, often within
one day", by 3 weeks the ratio OKT8:OKT4 was about 70:30, and that the
"change did not appear to be simply the preferential outgrowth of OKT8+
cells", but to a "possible change in phenotype of cultured human
lymphoblasts, from OKT4 to OKT8" (Burns et al., 1982). One year later in
1983, Zagury, an eminent HIV researcher (Zagury et al., 1983) and Gallo
collaborator, and his colleagues, selected normal human T cells for in
vitro cloning according to the expression of T4, T8 or T10 antigens on
individual cells. The individual cells were cultured in the presence of
TCGF (IL-2) "Preparations deprived of PHA", and "an irradiated lymphoid
cell filler- layer". They summarised their findings as follows: "Clones
were produced from each of these cells irrespective of the antigenic
phenotype of the parental cell. The cloned progeny manifested, in many
cases, shifts in antigen expression. Thus, T4+T8- cells gave clones
expressing predominantly T4-T8+ and vice versa. The clonal expression of
T4 and T8 seemed to be mutually exclusive. Antigenic shifts were recorded
also in clones derived from T4-T8-T10- cells, resulting in T10+ clones
which were also either T4+ or T8+ and from T4+T8-T10+ cloned cells
yielding clones of either T4+ or T8+ cells. Testing functional properties
we found that NK activity was mediated not only by T10+ cells but also, in
some cases, by T4+ and T8+ cells. Moreover, TCGF production, which may
reflect helper activity, was mediated not only by T4+ cells. Only the
cytotoxic (CTL) activity seems to be confined to the T8 phenotype. Thus,
it appears that T antigens, which seemed to be molecular markers of
differentiation, are not markers for terminal differentiation and do not
always reflect defined functional properties" (Zagury et al., 1983).
Given the in vitro evidence that:
(1) HIV is neither necessary nor sufficient for the observed decrease in
T4 cell numbers;
(2) T4 cells can change into T8 cells while the sum of T4 + T8 remains
constant;
(3) stimulation of T cells by PHA, ConA, radiation, PMA and polybrene all
of which are oxidising agents leads to "down regulation" of CD4 and change
of T4 to T8; and the evidence that:
(i) individuals from the AIDS risk groups are exposed to many oxidising
agents including well known mitogens;
(ii) in individuals at risk for developing AIDS the decrease in T4 cell
number is paralleled by an increase in T8 cells (decrease in the T4/T8
ratio), while the total T cell numbers remains constant;
(iii) in individuals belonging to the main AIDS risk groups the above
changes can be observed in the absence of HIV,
one must conclude that:
(a) the decrease in the T4 cell numbers and increase in T8 cell numbers in
"HIV infected" cultures and individuals is due to agents other than HIV;
HIV is neither necessary nor sufficient for the induction of the above
phenomenon;
(b) in vivo the above changes may not be due to a selective destruction of
T4 cells and increased proliferation of T8 cells, but loss of T4 surface
markers and acquisition of T8 surface markers.
T4 and the clinical syndrome
The HIV/AIDS researchers consider T4 decrease as being the "hallmark" and
"gold standard" of HIV infection and AIDS (Shaw et al., 1988; Levacher et
al., 1992). In fact, in the most recent (1992) CDC AIDS definition, an
AIDS case can be defined solely on serological, (positive HIV antibody
test), and immunological (T4 cell count less than 200 X 106/L), evidence
(CDC, 1992). The new definition also requires that "the lowest accurate,
but not necessarily the most recent, CD4+ T- lymphocyte count should be
used" to define an AIDS case (CDC, 1992). However, ample evidence exists
that T4 cell decrease can be induced by many factors, some trivial, such
as sun bathing and solarium exposure, a decrease which can persist for at
least two weeks after exposure has ceased (Hersey et al., 1983; Walker &
Lilleyman, 1983). T4 cell counts "can vary widely between labs or because
of a person's age, the time of day a measurement is taken, and even
whether the person smokes" (Cohen, 1992). That many factors can affect the
T4 cell number is reflected by their large variation in HIV positive
patients. In one such study, patient measurements repeated by one
laboratory within 3-days showed a "minimum CD4+ cell count of 118
cells/mm3 and a maximum CD4+ cell count of 713 cells/mm3" (Malone et al.,
1990). In the MACS, consisting of 4954 "homosexual/bisexual men", it was
stressed that physicians and patients should be "aware that a measured CD4
cell count of 300X106/L really may mean it is likely that the "true" CD4
cell state is between 178 and 505X106/L. Thus there is no certainty this
person's "true CD4" is less than 500X106/L or that it is greater than
200X106/L" (Hoover et al., 1992). It is important to note that these
variations were obtained despite the fact that the CD4 measurements were
undertaken in laboratories which "are carefully standardized in an ongoing
quality control program".
In a study (Brettle et al., 1993) which examined the impact of the 1993
CDC AIDS definition on the annual number of AIDS cases as compared to the
1987 definition, it was found that if the definition was based on:
(i) the "first of two consecutive CD4 cell counts < or equal to 200 X
106/L", the number of AIDS cases doubled;
(ii) one abnormal CD4 count, the number of AIDS cases trebled.
Researchers at the university of California at Los Angeles School of
Medicine found that 5% of healthy persons seeking life insurance had
abnormal T4 cells counts, and that "In a subgroup of patients, the low
T-cell numbers or ratios appear to be stable findings". They concluded:
"In the absence of a history of a specific infection or illness or major
abnormalities on a physical examination, it is not worthwhile to attempt
to find a specific cause for the abnormality of T- cell subsets...A
uniform approach to this problem throughout the medical community will
help alleviate patients' anxiety and reduce the concern of the insurance
industry about this relatively common problem" (Rett et al., 1988).
If LAS, ARC, and the AIDS indicator diseases such as KS and PCP are the
consequence of T4 cell depletion then all groups of people who have a low
T4 cell count, irrespective of cause, should have high frequencies of
opportunistic infections and neoplasms. Conversely, all patients with AIDS
indicator diseases should have abnormally low T4 cells.
In a study on the effects of blood transfusion on patients with
thalassaemia major, researchers at the Cornell university Medical Center
and the Sloan-Kettering Institute for Cancer Research observed decreased
T4 cell numbers and inverted T4/T8 ratios associated with the
transfusions, but no increase in KS or PCP, and concluded that "...studies
which define transfusion related AIDS on the basis of analyses with
monoclonal antibodies must be viewed with caution" (Grady et al., 1985).
Although patients with alcoholic liver disease do not develop KS, PCP and
other AIDS indicator diseases more often than usual, they have both immune
deficiency and positive HIV antibody tests leading researchers from the
Veterans Administration Medical Centre to stress the importance of
recognising these facts: "...lest these patients be falsely labelled as
having infection with the AIDS virus and suffer the socioeconomic
consequences of this diagnosis" (Mendenhall et al., 1986).
Patients who have malaria have severe immunoregulatory disturbances
including decrease in T4 cells. A significant number of these patients
also test positive for HIV but they do not develop the AID clinical
syndrome, leading Volsky et al to conclude, "exposure to HTLV-III/LAV or
the related retrovirus and the occurrence of severe immunoregulatory
disturbances may not be sufficient for the induction of AIDS" (Volsky et
al., 1986).
The MACS in the USA showed that "even in the absence of treatment, close
to 25, 15 and 10% of men were alive and asymptomatic 4, 5 and 6 years
after first CD4+ <200 X 106/L measurement" (Hoover, 1993). In the same
study comparing HIV positive individuals who within five years progressed
to AIDS (Group A) with that those who did not (Group B), it was found
that: "receptive anal intercourse both before and after seroconversion
with different partners was reported more frequently by men with AIDS. The
ratio of the differences in this sexual activity between groups A and B
was higher at 12 (2.3) and 24 (2.6) months after seroconversion than
before seroconversion (2.0)". It was concluded that "sexually transmitted
co-factors, preseroconversion and/or postseroconversion...augment (or
determine) the rate of progression to AIDS" (Phair et al., 1992). However,
since:
(a) sexually transmitted infectious agents are bi- directionally
transmitted, that is, from the active to the passive partner and
vice-versa;
(b) in the above study the only sexual act directly related to the
progression to AIDS was passive anal intercourse (unidirectionally);
one would have to conclude that the "co-factors that augment (or
determine)" progression to AIDS are non-infectious. These findings are in
agreement with the oxidative theory of AIDS which claims that both HIV
phenomena (RT, virus-like particles, antigen/antibody reactions,
"HIV-PCR") and AIDS are caused by the many oxidative agents (including
semen), to which the AIDS risk groups are exposed (Papadopulos-Eleopulos,
1988; Papadopulos-Eleopulos et al., 1989a; Papadopulos-Eleopulos et al.,
1992a; Papadopulos-Eleopulos et al., 1992b) [PCR=polymerase chain
reaction].
According to Canadian researchers, "In TB as well as in lepromatous
leprosy, an immunosuppressive state will frequently develop in the host.
This state is characterised by T lymphopenia with a decreased number of T
helper cells and an inverted T-helper/T-suppressor cell ratio
...immunosuppression induced by the infection with M.tuberculosis can
persist for life, even when TB is not progressive" (Lamoureux et al.,
1987). Yet these patients do not have high frequencies of KS, PCP or other
AIDS indicator diseases. In other words, decrease in T4 cells is not
sufficient for the AIDS indicator diseases to appear. This is also
supported by evidence from animal studies. Experimental depletion of T4
cells in mice used as models for systemic lupus erythematosus in humans
did not lead to increased frequencies of neoplasms, nor did mice "develop
infectious complications, even though they were housed without special
precautions". In fact mice with low T4 cell numbers had "prolonged life"
(Wofsy & Seaman, 1985) It is also of interest that despite the
indispensable role attributed to T4 and T8 lymphocytes in antibody
production (helper and suppressor respectively), AIDS patients in the
presence of low numbers of T4 cells and high numbers of T8 cells, have
increased levels of serum gammaglobulins, and are not
hypogammaglobulinaemic as might be expected. Also, although human
umbilical cord T-cells produce suppressor factors(s), the factor(s) is
produced by T8- (T4+) not T8+ cells (Cheng & Delespesse, 1986). Thus, T4
and T8 cells do not seem to possess the generally accepted functions
attributed to them.
According to the HIV theory of AIDS pathogenesis, "The Human
Immunodeficiency Virus (HIV), the etiologic agent of the acquired
immunodeficiency syndrome (AIDS), has the capability of selectively
infecting and ultimately incapacitating the immune system whose function
is to protect the body against such invaders. HIV-induced
immunosuppression results in a host defense defect that renders the body
highly susceptible to "opportunistic" infections and neoplasms" (Fauci,
1988). Decrease of T4 cells to approximately 200X106/L leads to the
development of "constitutional symptoms", and to less than 100X106/L to
"Opportunistic diseases" (Pantaleo et al., 1993). If this is the case
then:
1. In all individuals with "constitutional symptoms", OI and neoplasms,
the T4 cell number should be abnormally low;
2. The decrease in T4 cells should precede the development of the clinical
symptoms since: (a) the cause must precede the effect; (b) for many
neoplastic and infectious diseases, there is evidence that the diseases
themselves and the agents used to treat them may induce immune suppression
including decreased numbers of T4 lymphocytes and reversal of T4/T8
ratios.
This is not the case even for the most serious and characteristic of the
AIDS diseases, KS and PCP. In the MACS it was reported that:
(a) "...persistent generalised lymphadenopathy was common but unrelated to
immunodeficiency", and "Although seropositive men had a significantly
higher mean number of involved node groups than the seronegative men (5.7
compared with 4.5 nodes, p<0.005), the numerical difference in the means
is not striking".
(b) weight loss, diarrhoea, fatigue, fever, which constitute the "wasting"
syndrome, (which at present is an AIDS indicator disease), night sweats,
herpes zoster, herpes simplex (another AIDS indicator disease), oral
thrush, fungal skin infections and haematological abnormalities, were
present in both seronegative and seropositive individuals, although some
of them were present at higher frequencies in the latter group. A
relationship was found between thrush, anaemia, fever and neutropenia and
T4 cell deficiency. However, "the clinical abnormalities were considerably
better at reflecting concurrent CD4 lymphocyte depression than the low CD4
lymphocyte counts were at determining clinical involvement" (Kaslow et
al., 1987). These observations are just as compatible with the hypothesis
that T4 lymphocyte deficiency is the result and not the cause of the
observed clinical abnormalities.
KS, the main reason for which the retroviral hypothesis was put forward,
was initially postulated to be caused by infection of normal cells with
the retrovirus. When, late in 1984 it became clear that the KS cells were
not infected with HIV, it was generally accepted that the disease was
caused by HIV indirectly, that is, as a consequence of T4 cell decrease.
At present, it is generally believed that KS is caused by "a specific
sexually transmitted etiologic agent" (Beral et al., 1990; Weiss, 1993)
other than HIV, but "immune suppression (both in AIDS and in transplant
patients) is the dominant cofactor for subsequent disease" (Weiss, 1993).
However, unlike the Unites States CDC and most AIDS centres around the
world, for the Walter Reed Army Institute of Research "...the presence of
opportunistic infections is a criterion for the diagnosis of AIDS, but the
presence of Kaposi's sarcoma is omitted because the cancer is not caused
by immune suppression..." (Redfield & Burke, 1988) In a study by a group
of researchers from Amsterdam regarding the relationship between the T4
cell number and the development of the clinical syndrome, KS was excluded
"Because Kaposi's sarcoma may manifest at higher CD4+ lymphocyte counts
than other AIDS- defining conditions" (Schellekens et al., 1992). This is
not surprising since by the beginning of the AIDS era, the immune
surveillance hypothesis of carcinogenesis had been already refuted
(Kinlen, 1982). In fact, the presently available data indicate that KS in
all individuals, including gay men, may be caused by a non-infectious
agent (Papadopulos-Eleopulos et al., 1992a). Even in the early stages of
the AIDS era, it was reported that KS in gay men appeared following
corticosteroid administration (which was administered for diseases totally
unrelated to HIV or AIDS) and resolved when the drug was discontinued
(Schulhafer et al., 1987; Gill et al., 1989). Thus the HIV/AIDS hypothesis
cannot account for the very disease for which it was originally put
forward.
In a study of 145 patients, 97% of whom were homosexuals, with biopsy
proven PCP at St. Vincent's Hospital and Medical Centre, New York, 17% of
AIDS patients had a T4 cell count higher than 500/mm3, and a further 14%
between 301-500/mm3, "in addition, patients with T4-T8 ratio greater than
1.0 and those with total T4 lymphocyte counts greater than 500/mm3 cells
did not show improved survival compared with patients with abnormal
values....the degree of suppression did not influence mortality (Kales et
al., 1987). Researchers from the National Institute of Allergy and
Infectious Diseases and the National Cancer Institute, studied 100
HIV-infected patients "who had 119 episodes of pulmonary dysfunction
within 60 days after CD4 lymphocyte determinations". T4 cells were less
than 200X106/L before 46 of 49 episodes of PCP, 8 of 8 episodes of CMV
pneumonia, 7 out of 7 Cryptococcal neoformans pneumonia, 19 of 21 episodes
of Mycobacterium avium-intracellulare pneumonia, 6 of 8 [pulmonary] KS and
in 30 out of 41 non-specific interstitial pneumonia. However, "Before the
119 episodes of pulmonary dysfunction were diagnosed in this study, the
HIV- infected patients had manifested the following clinical HIV- related
disorders: no disorders (4 episodes), Kaposi's sarcoma without
opportunistic infections (68 episodes), life- threatening opportunistic
infection (44 episodes), other AIDS- related conditions (11 episodes)". In
addition before the diagnosis of the pulmonary episodes the patients had
received: "zidovudine (36 episodes), interferon (23 episodes), recombinant
interleukin-2 (3 episodes), cytotoxic chemotherapy (16 episodes),
dideoxycytidine (6 episodes), muramyl tripeptide (1 episode), suramin (6
episodes), heteropolyanion 23 (5 episodes), zidovudine plus interferon (5
episodes), nonablative bone marrow transplantation (4 episodes). Twenty-
two episodes occurred in patients who had been receiving neither
experimental therapy nor zidovudine" (Masur et al., 1989). These data may
be interpreted as showing that in some types of "pulmonary dysfunction",
most cases (but not all) appear to be preceded by a CD4 count <200X106/L.
However, given the well known fact that malignant neoplasms, infectious
diseases and the administration of chemotherapeutic agents may themselves
cause immunosuppression (Serrou, 1974; Oxford, 1980; Reinherz et al.,
1980; Rubin et al., 1981; Thomas, 1981; Weigle et al., 1983; Williams et
al., 1983; Kempf & Mitchell, 1985; Feldman et al., 1989), it is equally
plausible to argue that both "pulmonary dysfunction" and the low CD4 cell
counts observed in patients were the result of their recent past illnesses
and previous exposure to prescribed and illicit drugs and other factors.
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