Buggert, Marcus Nguyen, Son McLane, Laura M. Steblyanko, Maria Anikeeva, Nadia Paquin-Proulx, Dominic Del Rio Estrada, Perla M. Ablanedo-Terrazas, Yuria Noyan, Kajsa A. Reuter, Morgan Demers, Korey K. Sandberg, Johan A. Eller, Michael Streeck, Hendrik Jansson, Marianne Nowak, Piotr Sönnerborg, Anders H. Canaday, David Naji, Ali John Wherry, E. L. Robb, Merlin G. Deeks, Steven Reyes-Teran, Gustavo Sykulev, Yuri C. Karlsson, Annika R. Betts, Michael Cytolytic CD4+ T cells express high levels of T-bet and Eomes in blood. <p><b>(A)</b> Representative flow cytometry plots of Granzyme B and perforin expression in CD4+ T cells for an HIV-infected and–uninfected subject. The distribution of Granzyme B+perforin+ (red) and Granzyme B-perforin- (blue) CD4+ T cells are shown for T-bet and Eomes expression. <b>(B)</b> Frequency of perforin+ CD4+ T cells within the T-bet<sup>hi</sup>Eomes+ and T-bet<sup>dim/-</sup> population (left) and T-bet<sup>hi</sup>Eomes+ within the perforin+ or perforin- population for HIV-infected and–uninfected subjects. <b>(C)</b> Correlation between the frequency of perforin+ and T-bet<sup>hi</sup> CD4+ T cells. <b>(D)</b> Imagestream analysis on T-bet<sup>hi</sup> and T-bet<sup>dim</sup> CD4+ T cells. Overlays of fluorescent channels for DAPI (nuclear) and T-bet, showing where in the cells T-bet are localized. The frequency of nuclear, nuclear/cytoplasmic and cytoplasmic localization for T-bet<sup>hi</sup> and T-bet<sup>dim</sup> CD4+ T cells are shown in the before-after graphs. <b>(E)</b> tSNE plots based on 30,000 live CD4+ T cells that were merged from three HIV-uninfected subjects with detectable cytolytic CD4+ T cells. The tSNE clustering is based on CD45RO, CD27, CCR7, T-bet, Eomes, Granzyme A, Granzyme B and perforin expression intensity. The red gate indicates the identified “effector” cluster with overlapped expression of cytolytic markers as well as T-bet and Eomes. <b>(F)</b> Flow plots of MIP-1α production using media (NC) and aCD3-CD28 stimulations for T-bet<sup>hi</sup> and Eomes+ CD4+ T cells, as well as correlation between the frequency of T-bet<sup>hi</sup>Eomes+ and MIP-1α+ CD4+ T cells following aCD3-CD28 stimulations. Median and IQR are shown for all scatter plots and Mann-Whitney tests were performed to compare differences between groups; ***<i>P</i> < 0.001. A non-parametric Spearman test was used for the correlations analysis. All data are derived from the North-American cohort.</p> HIV replication;T-box transcription factors T-bet;transcriptional effector T cell properties;lack effector functions;IFN;cytolytic molecules;HIV elite controllers;LN;lymphoid tissues;chemokine 2018-04-13
    https://plos.figshare.com/articles/figure/Cytolytic_CD4_T_cells_express_high_levels_of_T-bet_and_Eomes_in_blood_/6141335
10.1371/journal.ppat.1006973.g001