Measuring VIRAL LOAD WITHOUT VIRUS: Where are the virions?
Posted by Henry Bauer on 2008/08/10
A continuing puzzle, at least for this lay person, is why HIV/AIDS researchers have never bothered to extract virions—whole particles of HIV—from HIV-positive people or from AIDS patients. Soon after “infection”, after all, the former are supposed to be teeming with virus, and AIDS victims are supposed to be full of virus (again) by the time opportunistic infections get a foothold; according to Fauci et al., there are then about 1,000,000 million and 100,000 “HIV RNA copies”, respectively, in each milliliter of plasma, each copy supposedly representing a virion:
Since primary infection and “acute viral syndrome” are often unaccompanied by any clinical symptoms—at best (or worst) mild flu-like signs or rashes—I had long thought that it would be unfair to chide mainstream researchers for failing to extract genuine virus at that stage. But, it turns out, some researchers have been able to carry out sophisticated studies of blood drawn during those critical initial weeks of primary infection.
Gasper-Smith et al. report on “Induction of plasma (TRAIL), TNFR-2, Fas ligand, and plasma microparticles after Human Immunodeficiency Virus Type 1 (HIV-1) transmission: Implications for HIV-1 vaccine design”, Journal of Virology 82  7700-10. They conclude that “Release of products of cell death and subsequent immunosuppression following HIV-1 transmission could potentially narrow the window of opportunity during which a vaccine is able to extinguish HIV-1 infection and could place severe constraints on the amount of time available for the immune system to respond to the transmitted virus”.
The researchers had been able to obtain from ZeptoMetrix Corporation of Buffalo (NY) “seroconversion panels” consisting of “sequential aliquots of plasma (range, 4 to 30 aliquots) collected approximately every 3 days during the time of acute infection with HIV-1”; they cite, for the availability of these seroconversion panels, Fiebig et al., “Dynamics of HIV viremia and antibody seroconversion in plasma donors: Implications for diagnosis and staging of primary HIV infection” , AIDS 17  1871-9.
Here, it seemed to me, had been an ideal opportunity to extract veritable whole particles of HIV generated during the acute initial infection. But the only mention of “virion” in the Gasper-Smith article is in this sentence: “While the average peak HIV-1 VL level was 1,421,628 copies/ ml, the average total MP peak level was 606,881,733/ml. Thus, at the times of maximum VL and MP levels, the average number of MPs was 427 times larger than the average number of virions”. “VL” of course is viral load. “MP” is not military police (or, as Lucas reminded me, Members of Parliament), it is “microparticles”:
“MPs are small membrane-bound vesicles that are released from the surface of apoptotic cells by exocytic or budding processes; . . . . MPs, which circulate in the blood under many clinical conditions, are part of a spectrum of subcellular structures that are released from cells and can be distinguished from exosomes . . . . MPs have immunomodulatory activities and can promote immune cell death; exosomes are also immunologically active, can suppress immune responses . . . , and have been reported to have been found at elevated levels in cases of chronic HIV-1 infection . . . . If elevations in levels of immunosuppressive molecules, coupled with early CD4+ T-cell death, occur early following HIV-1 transmission, then these events could potentially define a protected time during which HIV-1 is able to replicate while anti-HIV-1 T- or B-cell responses are suppressed” [emphases added].
Gasper-Smith et al. counted and extracted and studied the MPs by flow cytometry and electron microscopy. Why did they not also study HIV particles? Did the freezing and storing of the plasma destroy HIV virions while leaving MPs intact?
There were 427 times as many MPs as copies of RNA supposed to stem from HIV. MPs can “promote immune cell death”. How do we know that the CD4 cells supposedly killed by HIV weren’t killed by the MPs?
Though phrased rhetorically and left unanswered, I intend those questions to be taken quite seriously. If I wanted to be flippant or sarcastic, I might have commented once again on the peculiar penchant among HIV/AIDS researchers to imply that their measurements are accurate to an impossible number of significant figures when they report MPs of “606,881,733/ml”. That’s one of the drawbacks of the digital age, I suppose. In the good old days when we read measurements off scales with pointers, we weren’t tempted to write down meaningless numbers.
Perhaps Fiebig et al., cited by Gasper-Smith et al. for the brilliant idea of getting those stored samples from blood donors, had looked for whole particles of HIV?
“Because of the difficulty in obtaining blood samples representing early acute HIV infection from clinical patients, most patients do not come to medical attention until weeks to months after infection, we resorted to stored, frozen plasma collections from plasma donors, who unrelated to donating became infected with HIV, and were deferred from further donating. As plasma donors donate on average twice a week, and every donation is tested for HIV and held for 60 days before release, their archived samples provide a unique record of the infection from timepoints before viral exposure until seroconversion and beyond. . . . Plasma donations (600-800 ml) from source plasma donors were routinely collected at approximately twice weekly intervals and stored frozen at -20oC or less.”
Plenty of material to work with, it would seem—600 ml is well over a pint, and ought to contain many millions of HIV virions, at “1,421,628” per ml.
But, NO. In the Fiebig article, there’s not a single mention of “virion”. They used ELISA, p24 antigen, and HIV-1-RNA tests to determine how much “HIV” was present.
Is the failure to even try to extract virions somehow related to the fact that Gallo was more often able to “isolate” HIV from “pre-AIDS” patients than from those who actually had AIDS? Here’s from the Abstract of Gallo’s ground-breaking article that followed the press conference announcing discovery of the probable cause of AIDS:
“Peripheral blood lymphocytes from patients with the acquired immunodeficiency syndrome (AIDS) or with signs or symptoms that frequently precede AIDS (pre-AIDS) were grown in vitro with added T-cell growth factor and assayed for the expression and release of human T-lymphotropic retroviruses (HTLV)” (Gallo et al., Science 224  500-3).
That’s what Gallo means by “isolation”, as other rethinkers have often remarked. It’s not the commonly used meaning of the word, namely, “extraction” or “separation from”. And it’s not as though the “assaying” involved separating virions from those cultures, either.
“Retroviruses . . . were isolated from a total of 48 subjects including 18 of 21 patients with pre-AIDS, three of four clinically normal mothers of juveniles with AIDS, 26 of 72 adult and juvenile patients with AIDS, and from one of 22 normal male homosexual subjects”.
Why from more pre-AIDS than from actual AIDS patients?
The Abstract ends with “These results and those reported elsewhere in this issue suggest that HTLV-III may be the primary cause of AIDS” [emphases added].
From that modest suggestion, the dogma that HIV causes AIDS evolved without the benefit of direct isolation—extraction, separation—of whole infectious virions from even a single HIV-positive or AIDS-suffering person, or from plasma preserved from periods of “acute viral syndrome”.
This entry was posted on 2008/08/10 at 9:38 am and is filed under HIV skepticism, HIV/AIDS numbers. Tagged: acute viral syndrome, Andrew Conrad, Anthony Fauci, Barton F. Haynes, Bhupat D. Rawal, Charles F. Reich III, Charles Heldebrant, David J. Wright, David S. Pisetsky, Deanna M. Crossman, Eberhard W. Fiebig, Gene M. Shearer, George M. Shaw, Guido Ferrari, HIV never isolated, HIV virions, Janet S. Ottinger, John F. Whitesides, Kent J. Weinhold, Laura E. Jones, Li Qin, Lorraine Peddada, M. Anthony Moody, Mark Kaplan, Michael P. Busch, microparticles and HIV, Mikulas Popovic, Nadia Mensali, Nancy Gasper-Smith, Patricia E. Garrett, Richard Smith, Richard T. Schumacher, Robert Gallo, Sara E. Miller, Stephen G. Self, Steven G. Plonk, Steven H. Kleinman, Syed Z. Salahuddin, Thomas N. Denny, viral load and virions. You can follow any responses to this entry through the RSS 2.0 feed. You can leave a response, or trackback from your own site.