Strong infiltration of CD8+CXCR5+ T cells into HCC tumors predicts a better prognosis. Fresh samples were stained with anti-CD8, anti-CXCR5, anti-PD-1 and anti-ICOS antibodies. (A–B) Following gating of CD8+ T cells, the frequencies of CD8+CXCR5+ T cells from healthy PBMCs (n=20), and matched HCC tumor tissue, peritumoral liver tissue and PBMC samples (n=40) were analyzed. (A) One representative experiment is shown. (C) Association of tumor-infiltrating CD8+CXCR5+ T cells with microvascular invasion (n=25 for positive, n=15 for negative) is shown. (B–C) The data indicate the median with the interquartile range. (D) Patients were divided into two groups (Low/High) based on the median of the tumor-infiltrating CD8+CXCR5+ T cell percentages. The early recurrence rate was compared between the two groups using the log-rank test. (E–F) PD-1 and ICOS expression by CD8+CXCR5+ T cells differed among tumor tissue and matched peritumoral tissues and peripheral blood (n=19). The data indicate the median with the interquartile range. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 determined using the Mann-Whitney U test (B, C, E and F).