Reliable Estimation of CD8 T Cell Inhibition of In Vitro HIV-1 Replication

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  • Xu, Y.
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
  • Weideman, A.M.
    • Affiliation: Gillings School of Global Public Health, Department of Biostatistics
  • Abad-Fernandez, M.
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
  • Mollan, K.R.
    • Affiliation: Gillings School of Global Public Health, Department of Epidemiology
  • Kallon, S.
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
  • Samir, S.
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
  • Warren, J.A.
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
  • Clutton, G.
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
  • Roan, N.
    • Other Affiliation: University of California-San Francisco
  • Adimora, A.A.
    • Affiliation: Gillings School of Global Public Health, Department of Epidemiology
  • Archin, N.
    • Affiliation: School of Medicine
  • Kuruc, J.
    • Affiliation: School of Medicine
  • Gay, C.
    • Affiliation: School of Medicine
  • Hudgens, M.G.
    • Affiliation: Gillings School of Global Public Health, Department of Biostatistics
  • Goonetilleke, N.
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
Abstract
  • The HIV-1 viral inhibition assay (VIA) measures CD8 T cell-mediated inhibition of HIV replication in CD4 T cells and is increasingly used for clinical testing of HIV vaccines and immunotherapies. The VIA has multiple sources of variability arising from in vitro HIV infection and co-culture of two T cell populations. Here, we describe multiple modifications to a 7-day VIA protocol, the most impactful being the introduction of independent replicate cultures for both HIV infected-CD4 (HIV-CD4) and HIV-CD4:CD8 T cell cultures. Virus inhibition was quantified using a ratio of weighted averages of p24+ cells in replicate cultures and the corresponding 95% confidence interval. An Excel template is provided to facilitate calculations. Virus inhibition was higher in people living with HIV suppressed on antiretroviral therapy (n=14, mean: 40.0%, median: 43.8%, range: 8.2 to 73.3%; p < 0.0001, two-tailed, exact Mann-Whitney test) compared to HIV-seronegative donors (n = 21, mean: -13.7%, median: -14.4%, range: -49.9 to 20.9%) and was stable over time (n = 6, mean %COV 9.4%, range 0.9 to 17.3%). Cross-sectional data were used to define 8% inhibition as the threshold to confidently detect specific CD8 T cell activity and determine the minimum number of culture replicates and p24+ cells needed to have 90% statistical power to detect this threshold. Last, we note that, in HIV seronegative donors, the addition of CD8 T cells to HIV infected CD4 T cells consistently increased HIV replication, though the level of increase varied markedly between donors. This co-culture effect may contribute to the weak correlations observed between CD8 T cell VIA and other measures of HIV-specific CD8 T cell function.
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  • Article
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  • In Copyright
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  • Attribution 4.0 International
Journal title
  • Frontiers in Immunology
Journal volume
  • 12
Language
  • English
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  • Publisher
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  • National Institute of Allergy and Infectious Diseases, NIAID: P30AI050410, U01AI131310
ISSN
  • 1664-3224
Publisher
  • Frontiers Media S.A.

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