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. 2020 Dec 28;16(12):e1009177. doi: 10.1371/journal.ppat.1009177

Fig 6. Viral suppression of RT135T mutant virus by TN9-8T-specific CTL line.

Fig 6

A. Longitudinal sequence analysis at RT135 in an HLA-B*52:01+C*12:02+ individual with HIV-1, KI-638. Frequencies of amino acids at RT135 were determined by both bulk and deep sequencing. Both analyses showed similar results. The amino acids at RT135, their codon usage, and their frequencies are summarized by sampling date. B. Detection of TI8-, TN9-8V- and TN9-8T-specific CD8+ T cells in PBMCs from KI-638. These T cells were detected by TN9-8V/C1202 and TI8/B5201 tetramers or both TN9-8T/C1202 and TN9-8V/C1202 tetramers. C. Recognition of TN9-8T and its mutant peptides by TN9-8T-specific T cell line derived from KI-638. T-cell responses to 721.221-CD4-C1202 cells prepulsed with TN9-8T or its mutant peptides were analyzed by ICS. D. Binding of TN9-8V or TN9-8T tetramer to TN9-8T-specific T cell line. Data are shown as the mean and SD of triplicate assays E. Recognition of RT135 mutant virus-infected cells by a TN9-8T-specific T cell line. T-cell responses to 721.221-CD4-C1202 cells infected with NL43 (NL43-8I) or the mutant virus (NL43-8V, -8T or -8L) were analyzed by ICS. The frequencies of p24+ cells among 721.221-CD4-C1202 cells infected with NL43-8I, -8V, -8T, and -8L were 31.8%, 27.0%, 31.8%, and 28.5%, respectively. Representative flow cytometric results (left) and summarized results for the frequency of IFN-γ+ cells among CD8+ cells (right) is shown. F. Suppression of viral replication by TN9-8T-specfic CTL line, where E:T ratio was 1:1. The concentration of p24 Ag in the culture supernatants collected at Day 5 was measured by ELISA (left) and the percentage of suppression was calculated (right). G. Detection of TN9-8V- and TN9-8T-specific CD8+ T cells in PBMCs from 7 individuals infected with RT135T mutant virus. HLA-C*12:02-restricted TN9-8V and TN9-8T-specific CD8+ T cells were analyzed by using TN9-8T/C1202- or TN9-8V/C1202-tetramer. Statistical analysis was performed using a paired t-test (two-tailed). Data are shown as the means and SD of triplicate assays (C, D, E, and F).