Item #43: Can HIV be Isolated Without ‘Reactivating’ Latent Copies?

Generally speaking, HIV can be isolated only by “reactivating” latent copies of the virus, and then only with extraordinary difficulty. Viral load, one of the clinical markers for HIV, is not a measurement of actual, live virus in the body but the amplified fragments of DNA left over from an infection that has been suppressed by antibodies.

Farber states “HIV can be isolated only by 'reactivating' latent copies of the virus, and then only with extraordinary difficulty”. She supplies no reference.

This is false. Virus isolates are routinely made in clinical and basic research laboratories. It is true that more virus is produced by reactivating latent cells, but this is not what Farber is saying.

Virus ‘isolation’ actually consists of placing impure materials (such as a serum sample) into a cell culture (often of cancerous white blood cells) and adding stimulating chemicals. After several days the detection of proteins such as p24, the occurrence of reverse transcription or the presence of particles of the expected size and shape are taken as signs that a virus is present. More than that they are taken as signs that a specific virus, HIV, is present. For example, Robert Gallo wrote in 1987 that

“Continuous production of HTLV-III [HIV] is obtained after repeated exposure of parental HT cells to concentrated culture fluids containing HTLV-III harvested from short term cultured T-cells (grown with TCGF [T-Cell Growth Factor]) which originated from patients with pre-AIDS or AIDS. The concentrated fluids were first shown to contain particle-associated reverse transcriptase (RT)…Samples exhibiting more than one of the following were considered positive [for an HTLV-family virus]: repeated detection of a Mg++ dependent reverse transcriptase activity in supernatant fluids; virus observed by electron microscopy; intracellular expression of virus-related antigens detected with antibodies from sero-positive donors or with hyperimmune serum; or transmission of particles, detected by reverse transcriptase assays or by electron microscopic observation, to fresh human cord blood, bone marrow, or peripheral blood T-lymphocytes. All isolates not classified as either HTLV-I or HTLV-II by immunological or nucleic acid analysis were classified as HTLV-III [HIV]” [1]

“Evidence for the presence of HTLV-III [HIV] included: (i) viral reverse transcriptase (RT) activity in supernatant fluids; (ii) transmission of virus by coculturing T cells…; (iii) observation of virus by electron microscopy; and (iv) the expression of viral antigens…using serum from a patient positive for antibodies to HTLV-III…or antisera prepared against purified, whole disrupted HTLV-III…[using this methodology] we found HTLV-III in…13 of 43 of adult AIDS patients with Kaposi’s sarcoma, and 10 of 21 adult AIDS patients with opportunistic infections”

There are several published research papers that indicate that virus ‘isolation’ or cell culture is not always concordant with antibody evidence. This is not often found because virus culturing is not often done as it is a time-consuming process (this is one reason that most of these references are fairly old):

• “virus culture is time-consuming, expensive and does not directly measure virus titre” [2]
• “Virus has been repeatedly isolated in an additional small proportion of children (2.5%) who lost maternal antibody and have remained clinically and immunologically normal…virus isolation has been attempted in 163 [children who lost antibody]; in 10 (6.1%), positive results were obtained at least once” [3]
• “225 cultures of peripheral-blood lymphocytes from 133 seronegative men were performed, and HIV-1 was isolated in cultures from 31 men (23%). Of these men, four have seroconverted after being seronegative for 11 to 17 months after the initial isolation of the virus. The virus was not always isolated at every visit after the first successful isolation.” [4]
• “The study population consisted of...patients with AIDS and patients with AIDS-related complex…HIV was isolated at entry in 57% of the AZT group and 58% of the placebo group” [5]
• “In an early set of experiments, HTLV-III [HIV] was cultured from 48 subjects, including 18 or 21 patients with ARC [AIDS-related Complex, also known as pre-AIDS], 3 of 4 clinically normal mothers of children with AIDS, and [only] 26 of 72 adults and children with AIDS…Of interest is the finding, that while antibody to HTLV-III was more often associated with advanced disease, HTLV-III itself was more frequently cultured in ARC or newly diagnosed AIDS patients” [6]
• “fewer than 50% of patients with CD4 [immune cell] counts greater than 200 cells/microliter had positive plasma cultures” [7]
• “15 patients had no detectable virus from plasma culture at entry; 8 of these patients remained consistently culture negative, while the other 7 had virus isolated from plasma at some point during the 12 weeks of study…During the study, 9 [patients who did not have a culture obtained at study entry] had consistently negative cultures, 1 had consistently positive cultures, and 10 had intermittently positive cultures.” [8]
• “14 women seropositive for HTLV-III/LAV [HIV using ELISA antibody test] gave informed consent to culture of genital secretions and venous blood…from September, 1985, to December, 1985…[HIV] was cultured from cervical secretions in 4 of 14 women…[and] was detectable in blood in 7 of 13 women tested.” [9]
• “HIV was isolated from 49 of 156 immunoblot[ELISA]-positive [blood] donors (31%)…Virus was detected in cultures from 56 of 131 seropositive donors (43%) at the CDC” [10]

It is worth noting that David Baltimore, a high status researcher, wrote in 1985 that “In the last few years, the view that reverse transcription is solely a retroviral mechanism has been disproven”. [11] Another prominent researcher, Harold Varmus, wrote in 1982 that “a growing constellation of eukaryotic genetic elements – various pseudogenes and repetitive sequences – appear to depend upon reverse transcriptases of unknown provenance for their existence or amplification” [12].

1. Gallo RC et al. Method of continuous production of retroviruses (HTLV-III) from patients with AIDS and pre-AIDS using permissive cells. US Patent Office. 1987 Mar 24; 4,652,599.
2. Mortimer PP. The AIDS virus and the HIV test. Med Int. 1988; 56: 2334-9.
3. European Collaborative Study. Children born to women with HIV-1 infection: natural history and risk of transmission. Lancet. 1991; 337: 253-60.
4. Imagawa DT et al. Human immunodeficiency virus type I infection in homosexual men who remain seronegative for prolonged periods. N Engl J Med. 1989 Jun 1; 320(22): 1458-62.
5. Fischl MA et al. The Efficacy of Azidothymidine (AZT) in the Treatment of Patients with AIDS and AIDS-Related Complex. N Engl J Med. 1987 Jul 23; 317(4): 185-91.
6. Layon J et al. Acquired immunodeficiency syndrome in the United States: a selective review. Crit Care Med. 1986; 14(9): 819-27.
7. Saag MS et al. HIV viral load markers in clinical practice. Nat Med. 1996 Jun; 2(6): 625-9.
8. Meng TC et al. Combination therapy with recombinant human soluble CD4-immunoglobulin G and zidovudine in patients with HIV infection: a phase I study. J Acquir Immune Defic Syndr. 1995 Feb 1; 8(2): 152-60.
9. Vogt MW et al. Isolation of HTLV-III/LAV from cervical secretions of women at risk for AIDS. Lancet. 1986 Mar 8; 1(8480): 525-7.
10. Leitman SF et al. Clinical implications of positive tests for antibodies to human immunodeficiency virus type 1 in asymptomatic blood donors. N Engl J Med. 1989 Oct 5; 321(14): 917-24.
11. Baltimore D. Retroviruses and retrotransposons: the role of reverse transcription in shaping the eukaryotic genome. Cell. 1985 Mar; 40(3): 481-2.
12. Varmus HE. A growing role for reverse transcription. Nature. 1982 Sep 16; 299(5880): 204-5.