Figure 4. Effect of Entpd1 silencing on murine T cell responses after LCMV infection.

SMARTA CD4⁺ T cells transduced with the indicated shRNAmir retrovirus were transferred into B6 mice. Spleen cells were analyzed on day 8 (A–C) and day 3 (D) after LCMV infection. (A) Frequencies of Tfh (CXCR5^hiSLAM^lo) and Th1 (SLAM^hiCXCR5^lo) SMARTA cells. (B) Frequencies of SMARTA CD4⁺ T cells as percentage of total CD4⁺ T cells. (C) Frequencies of GC Tfh (CXCR5⁺BCL6⁺) cells. All data are shown as representative contour plots and summary data from 1 of 2 experiments with 4 to 5 mice in each group. (D) CXCR5⁺BCL6⁺ Tfh cells were analyzed and quantified by flow cytometry. Data from 1 experiment with 5 mice in each group. (E–H) OT-II CD4⁺ T cells transduced with the indicated retrovirus were transferred into CD4-knockout mice and analyzed 12 days after immunization with NP-OVA in alum. (E) Frequencies of FAS⁺GL7⁺ cells as percentage of total B cells. (F) Histological analysis of spleens from immunized mice. Representative images of B220, CD3, and GL7 staining and comparison of GC areas in mice reconstituted with control and shEntpd1 OT-II T cells. Scale bars: 50 μm. (G) Frequencies of CD138⁺IgD^– plasma cells. Representative contour plots and summary data from 1 experiment with 5 mice in each group. (H) NP-OVA–specific IgG titers. Data shown as mean ± SEM were compared by unpaired 2-tailed t test. *P < 0.05; **P ≤ 0.01; ***P ≤ 0.001. NS, not significant.