Monday, June 08, 2009

HIV Kills T cells...and doesn't need help to do it

Recently I've had cause to want to post again. Two things: an amusing comparison, and the an alarming take-over of a once-useful Society.

A certain Clark Baker (one-time cop and marine by his own account - a good reason to brush up on my Karate for self defence if ever there was it also appears his 'retirement' from the LAPD may have been spurred by an assault charge made by an immigrant) has started making noise and touting the same old disproven denialist lies (such as HIV is a harmless retrovirus, originally stated by Duesberg based on Duesberg's extensive personal experience working on HIV in the lab - ie none). Baker went one step further though. Taking advantage of what sounds like fundamental apathy in the society, he managed to infiltrate the Semmelweiss Society International (SSI) with other AIDS denialists, gain control of their bank account, and throw out the non-denialist board members. The SSI's remit was certainly honorable enough - to protect medical practitioners who are wrongfully accused of malpractise, but it has certainly lost its way. The takeover is all outlined in court documents filed against Baker which I have read. This infiltration certainly explains why the SSI saw fit to 'honor' Duesberg and Farber with whistle-blower awards.

The SSI website is now filled with the slightly hysterical, ill-informed, self-grandiosing propaganda that passes for AIDS denial these days.

The sad thing is that AIDS denial is no different from the Fiber One breakfast cereal. WTF I hear you say? Consider this:

The wife here is pissed, because "everyone knows" that foods high in fiber don't taste nice, therefore since Fiber One tastes nice, it can't have much fiber in it.

This is no different from Duesberg stating that "everyone knows" that retroviruses don't kill cells, and since HIV is a retrovirus, it can't kill cells.

The Fiber One commercials are funny. AIDS denial is resposible for killing hundreds of thousands of people.

Baker, on one of his recent rants, asks for proof that HIV kills T cells. Well, I pointed this out the AIDS denialists over a decade ago, but keeping up with the literature was never a strong point of AIDS denialism (denying the current science is what makes then denialists, after all). Incidentally this bit of research also explains a comment made by Montagnier (one of the original discoverers of HIV) where he states that a co-factor, along with HIV, may be responsible for AIDS - suggesting that HIV by itself might be relatively harmless. This, if it were true, wouldn't necessarily detract from the HIV->AIDS hypothesis, but it might have had an impact on how to treat or prevent AIDS, so it was well worth looking into.

Specifically, in this 1992 paper by Montagnier, they state that
Following infection of lymphoblastoid (CEM) or promonocytic (U937 and THP1) cell lines with HIV-1 cytopathic effect was observed only in association with mycoplasmal contamination. Moreover, HIV-1 infection of U937 cells after experimental inoculation with a human isolate of M. fermentans led to pronounced cell killing. We have verified that this effect is not merely an artifact caused by arginine and/or glucose depletion in the cell culture medium. These results confirm that mollicutes, in particular M. fermentans, are able to act synergistically with HIV-1 to kill infected cells in some in vitro systems.
He had noticed that doxycycline could protect against HIV-induced cytotoxicity back in 1990. The mycoplasma detected here is a common problem in laboratory cell culture systems - they are notorious for upsetting experiments. I wonder if this notoriety came about because of results like this. It is not surprising that a mycoplasma, or some other co-factor, might be considered in AIDS causality, and the denialists are all over results like this as proof that HIV is harmless. But Montagnier only looked for cytopathic effect - i.e. what do the cells look like under the microscope. Look at this paper, from a couple of years afterwards. I'll post the abstract - basically they showed that the cell line Montagnier used (CEM in his original 1990 and followup 1992 paper) was not a pure line - it was partly CD4+ and partly CD4-. HIV can only infect CD4+ cells as that is what its envelope proteins (gp120 and gp41) bind to. It will come as no surprise to those who actually follow HIV science that in CEM lines, all the CD4+ cells get killed, but the CD4- cells grow to fill in the gaps (i.e. no cytopathic effect for Montagnier to observe).

CEM cells were infected with three HIV-1 non syncytium-inducing (NSI) strains obtained from AIDS patients or seropositive individuals. The surviving cells were followed for several months in the persistently infected cultures designated 65870/CEM, 65871/CEM and 3929/CEM, and analyzed for virus expression using light and electron microscopy, immunofluorescence, reverse transcriptase assay, polymerase chain reaction amplification (PCR), nucleic acid hybridization and flow cytometry. The virus isolates induced relatively few syncytia and other cytopathic effects in the corresponding cell lines and the number of cells positive for virus expression never rose above 44%. Distinct peaks of antigen-positive cells were obtained, coincident with high levels of reverse transcriptase activity. The cultures were strongly resistant to superinfection by laboratory strain Lai, with the exception of 65870/CEM which expressed HIV antigens in up to 15% of the cells for a few days. However, cell lysis was minimal in all cases. After long-term cultivation of the three cultures, no antigen-positive cells were detected and no trace of virus expression could be observed. The remaining cells consisted entirely of CD4-negative cells. PCR analyses indicated that cells harboring a provirus were progressively eliminated from the cultures, leaving only virus-free cells. In this system, cells carrying a latent provirus survive for a limited period of time before virus activation induces cell lysis. These results suggest that at least three types of cells exist in the CEM cell line: CD4-positive cells which are rapidly killed by the virus, a second type harboring a latent viral genome after the infection and which grow normally until activation of the resident genome by external or internal signal(s), and a third type which represents rare CD4-negative cells present in the initial CEM population and which are selected for by the NSI isolates. This is the first study documenting specific interactions between NSI strains of HIV-1 and distinct subpopulations of CEM cells grown as a single cell culture.

This is a great example of how papers that apparently 'disprove' a hypothesis actually confirm it if you look at the science in greater detail.

To wrap up the story, it's well known that monocytes/macrophages like the U937 and THP-1 lines are responsible for longterm infection with HIV, the "reservoir" of infection that is hard to kill. But this makes sense - HIV doesn't deplete CD4+ macrophages to cause AIDS, so HIV wouldn't be expected to kill monocyte cell lines in the lab...unless perhaps the cultures were contaminated with mycoplasma... Maybe Duesberg was half-right.

In any case, just as Fiber One is a tasty source of fiber, HIV kills cells.

This whole story is a great example of why AIDS denialism has moved from a legitimate question-asking exercise, through inexplicable stubborness and willfull ignorance, to downright evil. When AIDS denial is touted by homophobic, ignorant, arrogant people who (allegedly) lie and steal in order to further their devastating agenda, then it's really fallen about as far as it can go.


Disclaimer: I have never bought Fiber One, nor do I endorse them. I have however worked with HIV in the lab in short and long-term culture systems and witnessed its cytopathic effect. I use the word "allegedly"above as Baker is so far accussed, but not proven, to be a homophobic, racist, ignorant, arrogant, liar and thief. However, anyone who calls AIDS patients "a small group of promiscuous, addicted, nitrite-huffing, gonorrheal and syphilitic bath house veterans" probably needs no further said of them.

Lemaitre M. Guetard D. Henin Y. Montagnier L. Zerial A. Research in Virology. 141(1):5-16, 1990 Jan-Feb Protective activity of tetracycline analogs against the cytopathic effect of the human immunodeficiency viruses in CEM cells.

M Lemaître, Y Henin, F Destouesse, C Ferrieux, L Montagnier, and A Blanchard
Infect Immun. 1992 March; 60(3) 742–748. Role of mycoplasma infection in the cytopathic effect induced by human immunodeficiency virus type 1 in infected cell lines.

Yelle et al Arch Virol 1994;139(1-2):155-72 "Analysis of long- term viral expression in CEM cells persistently infected with non syncytium-inducing HIV-1 strains.