Phenotypic, cytolytic and transcriptional differences between LN and blood CD4+ T cells in HIV-infected and -uninfected individuals. Marcus Buggert Son Nguyen Laura M. McLane Maria Steblyanko Nadia Anikeeva Dominic Paquin-Proulx Perla M. Del Rio Estrada Yuria Ablanedo-Terrazas Kajsa Noyan Morgan A. Reuter Korey Demers Johan K. Sandberg Michael A. Eller Hendrik Streeck Marianne Jansson Piotr Nowak Anders Sönnerborg David H. Canaday Ali Naji E. John Wherry Merlin L. Robb Steven G. Deeks Gustavo Reyes-Teran Yuri Sykulev Annika C. Karlsson Michael R. Betts 10.1371/journal.ppat.1006973.g003 https://plos.figshare.com/articles/figure/Phenotypic_cytolytic_and_transcriptional_differences_between_LN_and_blood_CD4_T_cells_in_HIV-infected_and_-uninfected_individuals_/6141341 <p><b>(A)</b> Flow cytometry plots (HIV-infected CP) and scatter plots for naïve and memory subsets of LN and peripheral blood (PB) CD4+ T cells in HIV-infected and -uninfected subjects. <b>(B)</b> Flow cytometry plots (HIV-infected CP) showing the lack of T-bet<sup>hi</sup>Eomes+ CD4+ T cells in LNs. Corresponding scatter plots demonstrating the frequency of T-bet<sup>hi</sup> cells of memory (non-naïve) CD4+ T cells (top) and frequency of Eomes+ cells of T-bet<sup>hi</sup> CD4+ T cells (bottom) for matched LN and PB. <b>(C)</b> Flow plots (HIV-infected CP) showing the lack of Granzyme B+perforin+ CD4+ T cells in LNs and scatter plots with the frequency of LN and PB perforin+ cells of memory CD4+ T cells (top). Frequencies of Granzyme B+ cells of perforin+ CD4+ T cells (bottom) for matched LN and PB. <b>(D)</b> Flow plots (HIV-infected CP) and scatter plots showing the distribution of CD27+ cells within the Granzyme B+ CD4+ T cell compartment for matched LN and PB. <b>(E)</b> The distribution of different populations in the tSNE space is based on 30.000 live CD4+ T cells that were merged from LN and PB from a HIV-infected CP with detectable levels of cytolytic cells in the PB and LN Tfh cells. The tSNE clustering is based on CD45RO, CD27, CCR7, T-bet, Eomes, Granzyme B, perforin, CXCR5 and PD-1 expression on gated bulk CD4+ T cells. The naïve cluster (green) is based on high CCR7 and low CD45RO intensity; the Tfh cluster (red) on high intensity of PD-1 and CXCR5; and the effector cluster (orange) on high T-bet and perforin expression intensity. After separating out the merged LN and PB single CD4+ T cell data, a lack of Tfh cells was apparent in PB and effector CD4+ T cells in the LN (lower right tSNE plots). Median and IQR are shown for all scatter plots. Mann-Whitney tests were performed to compare differences between two unmatched groups, and Wilcoxon matched-pairs single rank tests between matched samples; *<i>P</i> < 0.05, **<i>P</i> < 0.01 and ***<i>P</i> < 0.001. All data-points are derived from the North-American and Mexico cohort.</p> 2018-04-13 17:45:21 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