Seasonal influenza vaccines induce ICOS+CXCR5+ cells in blood. (A) Gating strategy for the analysis of blood CXCR5+CD4+ T cell subsets. (B) Percentage of ICOS+ cells within CD4+ T cells. Fresh blood samples were obtained from a healthy adult cohort (n = 12) before and after vaccination with trivalent seasonal influenza vaccine (Fluzone 2009/2010). The right panel shows data from six adults who received injection of normal saline. One-way analysis of variance (ANOVA). ***P < 0.001, paired t test. (C) A representative result of expression of ICOS on CXCR5+CD4+ T cells on day 7 after influenza vaccination or injection of normal saline. (D) Percentage of ICOS+ cells within CXCR5+ and CXCR5−CD4+ T cell compartments before and after influenza vaccination. Paired t test. n.s., not significant.