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. Author manuscript; available in PMC: 2020 Sep 1.
Published in final edited form as: Transplantation. 2019 Sep;103(9):1809–1820. doi: 10.1097/TP.0000000000002683

Figure 4. Suppression of alloantibody production by alloprimed CXCR5+CD8+ T cells is accompanied by a reduction in the quantity of Germinal Center B cells and CD4+ TFH cells.

Figure 4.

WT mice were transplanted with FVB/N hepatocytes (day 0) and on day 5 received adoptive transfer (AT) of sorted alloprimed CXCR5+CXCR3−CD8+ or CXCR3+CXCR5−CD8+ T cells. On day 14 post transplant, recipient splenocytes were analyzed for the number of germinal center (GC) B cells and CD4+ TFH by flow cytometry. A) GC B cells (GL-7+B220+) were analyzed by gating on lymphocytes. B) Recipients which received AT of CXCR5+CXCR3−CD8+ T cells exhibited significantly reduced number of GC B cells (22,000±2,000 per million splenocytes; n=5) compared to WT recipients without AT (38,000±2,000 per million splenocytes; n=5, p=0.001 signified by “*”). In contrast, the number of GC B cells was not significantly altered in recipients which received AT of CXCR3+CXCR5−CD8+ T cells (41,000±3,000 per million splenocytes; n=6, p=ns) compared to control recipients without CD8+ T cell transfer. C) CD4+ TFH cells were analyzed by gating on lymphocytes, CD4+ T cells, and PD-1+CXCR5+ cells. D) Recipients which received AT of CXCR5+CXCR3−CD8+ T cells exhibited significantly reduced number of IL-4+IL-21+ CD4+ TFH cells (4,400±100 per million splenocytes; n=5, p=0.003 signified by “*”) compared to WT recipients without AT (9,300±800 per million splenocytes; n=5). AT of CXCR5+CXCR3−CD8+ T cells was associated with a reduced number of IL-4+ CD4+ TFH cells (2,400±100 per million splenocytes; n=5, p=0.04) compared to control recipients without CD8+ T cell transfer (4,600±500 per million splenocytes). In contrast, the number of IL-4+ (4,600±300 per million splenocytes; n=6) and IL-4+IL-21+ CD4+ TFH cells (8,100±500 per million splenocytes; n=6) was not significantly altered after AT of CXCR3+CXCR5−CD8+ T cells compared to the control group with no CD8+ T cell transfer (p=ns for both IL-4+ and IL-4+IL-21+ CD4+ TFH cells). Error bars indicate standard error from duplicate experiments.