Figure 1
Phenotype of human CD40L+ CD8+ T cells. (A) The left dot-plot shows representative gates for ex vivo–sorted CD3+ CD8+ CD4− T-cell subsets based on CCR7 and CD45RA expression (CCR7+ CD45RA+ naive, CCR7+ CD45RA− central memory, CCR7- CD45RA− effector memory, or CCR7− CD45RA+ effector cells). The isolated cells were stimulated with P/I in the presence of αCD40 antibodies and were subsequently assessed for CD40L expression among activated CD69+ cells. (B) The frequencies of CD40L+ CD8+ T-cell subsets are shown as the median ± standard error of the mean (SEM) (n = 13); the Friedman test and Dunn’s post hoc test were used (***P < .001). (C) P/I-stimulated CD8+ T cells were analyzed for their capacity to coexpress CD40L with CD27, CD28, CD127, or CD57. (D) Ex vivo–sorted human memory CD8+CD45RA− T cells were stimulated with P/I and subsequently separated into CD40L+ and CD40L− fractions. Proliferation after 7 days in vitro culture with and without IL-2 and IL-12 was assessed by CFSE dilution and capability to re-express CD40L after 6-hour P/I stimulation. (C-D) One representative experiment out of 3 to 5 is shown.

Phenotype of human CD40L+ CD8+ T cells. (A) The left dot-plot shows representative gates for ex vivo–sorted CD3+ CD8+ CD4 T-cell subsets based on CCR7 and CD45RA expression (CCR7+ CD45RA+ naive, CCR7+ CD45RA central memory, CCR7- CD45RA effector memory, or CCR7 CD45RA+ effector cells). The isolated cells were stimulated with P/I in the presence of αCD40 antibodies and were subsequently assessed for CD40L expression among activated CD69+ cells. (B) The frequencies of CD40L+ CD8+ T-cell subsets are shown as the median ± standard error of the mean (SEM) (n = 13); the Friedman test and Dunn’s post hoc test were used (***P < .001). (C) P/I-stimulated CD8+ T cells were analyzed for their capacity to coexpress CD40L with CD27, CD28, CD127, or CD57. (D) Ex vivo–sorted human memory CD8+CD45RA T cells were stimulated with P/I and subsequently separated into CD40L+ and CD40L fractions. Proliferation after 7 days in vitro culture with and without IL-2 and IL-12 was assessed by CFSE dilution and capability to re-express CD40L after 6-hour P/I stimulation. (C-D) One representative experiment out of 3 to 5 is shown.

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