Failure of dideoxynucleosides to inhibit human immunodeficiency virus replication in cultured human macrophages.

Richman, Douglas D.; Kornbluth, Richard S.; Carson, Dennis A.

doi:10.1084/jem.166.4.1144

Cited by 142 publications

(55 citation statements)

References 24 publications

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“…3 A) . The value of 25 nM AZT for 50% inhibition of virus production is in accord with inhibitory concentrations measured by p24 production (29), plaque formation (23), or cell-free reverse transcriptase activity (30) .…”

Section: Resultsmentioning

confidence: 63%

Reinfection results in accumulation of unintegrated viral DNA in cytopathic and persistent human immunodeficiency virus type 1 infection of CEM cells.

Pauza

Galindo

1990

The Journal of Experimental Medicine

Self Cite

105

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SummaryHigh levels of unintegrated viral DNA accumulate during human immunodeficiency virus type 1 (HIV 1) infection of CEM T cells. Reinfection of already infected cells is required to attain these levels and reinfection also promotes the development of HIV-induced cytopathology. Rates of virus production, however, are independent of the accumulation of unintegrated viral DNA . Neutralizing antibody added soon after infection reduced viral DNA levels without appreciably affecting the production of cell-free viral p24 antigen or reverse transcriptase activity. Only 50 pM AZT were required to reduce the accumulation of unintegrated viral DNA by 50% in contrast to the 25 nM required to inhibit virus production by 50%. Cytopathology,-as measured by number of syncytia in infected cell cultures, was correlated with highly elevated levels of unintegrated viral DNA . The minimal levels of unintegrated viral DNA present constitutively in the persistently infected HCEM cell line were consonant with the absence of cytopathic effects in these cells. These data demonstrate that inhibiting the reinfection of already infected cells modulates cytopathic HIV-1 infection to a form that is persistent and noncytopathic.Depletion of CD4 T lymphocytes is an important feature of the natural history of HIV-1 infection (1, 2). Reduced numbers in this cell population predict disease progression (1-3) and are likely to contribute to immunodeficiency (4) . HIV-1 infection in vitro is rytopathic for T lymphocytes (5-7) and T lymphoid cell lines (8,9). This cytopathology occurs by the formation of syncytia when infected and uninfected CD4+ cells are admixed (8, 9) and direct cell killing by a mechanism independent of cell fusion (10) . Based on the cytopathic effects of HIV-1 infection of T cells in vitro, this mechanism was proposed to be a crucial feature of viral pathogenesis in AIDS (6,5,11, 12) . In this study, we have examined the role of reinfection and viral DNA accumulation in cytopathic and persistent HIV-1 infection of the CEM cell line.Host-pathogen interactions leading to cytopathology have been characterized extensively in avian retrovirus models. The accumulation of high levels of unintegrated viral DNA is a recognized feature of rytopathic infection by these retroviruses (13, 14) and was also shown to be an important marker of pathogenesis in the case of feline leukemia virus infection of cats (15) and equine infectious anemia virus infection of horses (16).Unintegrated viral DNA is generated by reverse transcription of infecting viral RNA . A linear viral DNA molecule 1035 flanked at both ends by long terminal repeat (LTR)t sequences (17) is generally found to be the most abundant form of unintegrated viral DNA . Intramolecular recombination between the LTR sequences then generates a circular molecule with one LTR that is of intermediate abundance . A circular DNA form with two LTR seems to arise via direct ligation of the ends of a linear molecule and is the least abundant species . Importantly, these DNA species are pro...

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Section: Resultsmentioning

confidence: 63%

Reinfection results in accumulation of unintegrated viral DNA in cytopathic and persistent human immunodeficiency virus type 1 infection of CEM cells.

Pauza

Galindo

1990

The Journal of Experimental Medicine

Self Cite

105

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“…The low level production of HIV from the latter cells is also an ideal sytem in which to investigate antibody enhancement of HIV uptake through Fc and complement receptors (Robinson et al, 1988;Takeda et al, 1988), as occurs with flaviviruses, alphaviruses, bunyaviruses, rhabdoviruses and reoviruses (Halstead & O'Rourke, 1977;Porterfield, 1986). The marked differences in productive HIV infection of fresh blood monocytes and plasticadherent macrophages must also be considered in functional studies of these cells, including HLA class II expression (Petit et al, 1987), antigen presentation (Lyerly et al, 1987;LeSane et al, 1988), response to or induction of cytokines (Hammer et al, 1986;Folks et al, 1987) and effect on antiviral function (Richman et al, 1987). Some studies suggesting that there is little effect on these functions may have been biased by the small percentage of macrophages infected with HIV.…”

Section: Discussionmentioning

confidence: 99%

Variations in CD4 Expression by Human Monocytes and Macrophages and Their Relationship to Infection with the Human Immunodeficiency Virus

Kazazi

Mathijs

Foley

et al. 1989

Journal of General Virology

134

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SUMMARYThe expression of CD4 antigen on the surface of LeuM3-positive human blood monocytes was found to be variable with 65 to 90~ of cells from 46 normal human volunteers being positive by dual staining flow cytometry. When monocytes adhered to plastic (but not when cultured on Teflon), a marked 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 immunofluorescence. Conversely an increasing proportion of adherent cells expressed LeuM3 and OKM5 surface antigens over the 5 days. CD4 mRNA levels were measured by slot-blot and Northern hybridization, and total cellular CD4 protein levels by immunoprecipitation. Both cellular mRNA and 'CD4 levels remained constant throughout the 5 day period but membrane CD4 protein levels were greatly reduced indicating that the down-regulation of CD4 was posttranslational. Infection with two of six fresh human immunodeficiency virus (HIV) isolates showed different kinetic patterns when tested on purified monocytes recently adhered to plastic and macrophages adherent for 5 days. HIV antigen and reverse transcriptase levels in infected monocyte cultures remained high for 3 to 4 weeks before detachment and necrosis of the cells occurred. Infection of macrophages generated much lower levels of antigen and reverse transcriptase which declined to very low or undetectable levels over 2 weeks, leaving persisting viable macrophages. One week after infection HIV nucleic acid was detected in 69 + 7~ of monocytes and 6 + 3~ of macrophages by in situ hybridization. Blocking experiments with anti-Leu3a monoclonal antibody suggested that HIV infection of 5 day adherent macrophages occurred mainly by a mechanism other than binding to CD4.

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“…Macrophages are a cellular target of HIV infection (Meltzer et al, 1990) and play a central role in the pathogenesis of AIDS because of their long-term persistance as cellular reservoirs of infection. Macrophages have low levels of dideoxycytidine kinase, and hence phosphorylate ddC at reduced rates compared to other cells (Richman et al, 1987;Perno et al, 1988). However, as suggested by Perno et al (1988), this reduced anabolic phosphorylation does not necessarily mean that ddC is not effective in inhibiting HIV replication in macrophages.…”

Section: Introductionmentioning

confidence: 90%

“…Macrophages have very low levels of deoxycytidine kinase (Richman et a/., 1987;Perno et el., 1988), but 2',3'-dideoxycytidine, a nucleoside analogue phosphorylated by this enzyme, has been reported to be effective in inhibiting HIV-1 replication (Perno et et.. 1987).…”

Section: Discussionmentioning

confidence: 99%

Inhibition of HIV-1 and LP-BM5 Replication in Macrophages by Dideoxycytidine and Dideoxycytidine 5′-Triphosphate

Magnani

Casabianca

Rossi

et al. 1995

Antivir Chem Chemother

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The antiviral agent 2′,3′-dideoxycytidine (ddC) has been shown to be active against HIV-1 infectivity. However, conflicting results have been reported concerning its efficacy in macrophages. Because macrophages possess low levels of the kinase(s) responsible for ddC phosphorylation, we investigated the ability of ddC and 2′,3′-dideoxycytidine 5′-trisphosphate (ddCTP) to suppress HIV-1 and LP-BM5 replication in these cells. Retrovirus replication was only partially inhibited in the two systems investigated by a high (1 μM) ddC concentration. The direct administration of ddCTP, using autologous red blood cells as a delivery system, was found to inhibit HIV-1 and LP-BM5 replication by more than 90% in macrophages without affecting major cell functions. These data, together with those already reported for FIV [Magnani et al. (1994) AIDS Res Hum Retroviruses 10: 1179-1186], suggest that the anabolic phosphorylation of ddC is an important determinant of its anti-HIV activity and that pharmacological interventions that modulate ddC metabolism may be useful for improving its antiretroviral activity in macrophages.

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Failure of dideoxynucleosides to inhibit human immunodeficiency virus replication in cultured human macrophages.

Cited by 142 publications

References 24 publications

Reinfection results in accumulation of unintegrated viral DNA in cytopathic and persistent human immunodeficiency virus type 1 infection of CEM cells.

Reinfection results in accumulation of unintegrated viral DNA in cytopathic and persistent human immunodeficiency virus type 1 infection of CEM cells.

Variations in CD4 Expression by Human Monocytes and Macrophages and Their Relationship to Infection with the Human Immunodeficiency Virus

Inhibition of HIV-1 and LP-BM5 Replication in Macrophages by Dideoxycytidine and Dideoxycytidine 5′-Triphosphate

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