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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: Virology. 2014 Jul 4;0:175–188. doi: 10.1016/j.virol.2014.06.008

Figure 3. GrzB production by CCR5+ activated naïve CD4, Th1, and Th17 cells.

Figure 3

(A) Constitutive upregulation of CCR5 by naïve CD4 T cells. Naïve CD4 T cells were purified from peripheral blood and cultured in complete RPMI medium without IL2 for 3–6 days (dotplots are representative of 3–4 experiments). (B–D) GrzB and HIV production by activated naïve CD4 T cells. Purified naïve CD4 T cells were stimulated with IL2 only or CD3/CD28 costimulation+IL2 for 3 days. Cells were then uninfected or infected with HIV for 1 day, washed, and cultured with IL2 for 6 days. GrzB expression by uninfected CCR5+ differentiated naïve CD4 T cells (B), and extracellular GrzB (C, one representative ELISA) and p24 (D, means±sem, n=3) of differentiated naïve CD4 T cells after 6 days post-infection culture. (E–F) Extracellular GrzB and p24 of Th1 or Th17 cells 5 days post-infection +/− IL2 (means±sem, *p<0.01, n=4). (G–H) GrzB coexpression with IFNγ by CCR5+ Th1 cells. Th1 cells were cultured in IL2 medium for 5 days, then stimulated with PMA/IO+GolgiPlug for the final 5hrs and stained by flow cytometry (*p<0.05 compared to respective CCR5− IFNγ/GrzB cells, n=4). (I–J) GrzB coexpression with IL17A by CCR5+/CCR6+ Th17 cells. Th17 cells were cultured in IL2 medium for 5 days, then stimulated with PMA/IO+GolgiPlug for the final 5hrs and stained by flow cytometry (*p<0.05 compared to respective CCR5−/CCR6− IL17A/GrzB cells, n=3).