Figure 1.

Antiprogrammed cell death protein 1 (anti-PD-1) treatment induces circulating T follicular helper cells (cTfh) in non-small cell lung cancer (NSCLC)-bearing mice. (A) Anti-PD-1 therapy reduces tumor development in KLN205 murine model of NSCLC. Graph represents tumor development in isotype-treated or anti-PD-1-treated mice (n=9) estimated as percentage of tumor size at the onset of therapy, measured by bioluminescence. (B) Quantification of follicular helper T cells (Tfh) as percentage of CD4+ T cells in peripheral blood (isotype n=29 and anti-PD-1 n=26), spleen (isotype n=18 and anti-PD-1 n=15) and draining lymph nodes (isotype n=8 and anti-PD-1 n=9) from isotype-treated and anti-PD-1-treated mice. (C) Plots show representative CXCR5 and PD1 surface staining of Tfh lymphocytes from peripheral blood and spleen of anti-PD-1-treated and isotype-treated mice. Percentages of each subpopulation are indicated. (D) Percentage of CD38, CXCR3 and CCR7 expression in Tfh from peripheral blood and spleens of anti-PD-1-treated or isotype-treated mice; n=9 for peripheral blood and n=12 for spleen of isotype-treated mice, and n=15 for both peripheral blood and spleen of anti-PD-1-treated mice. (E) Representative histograms of surface markers showed in (D). (F) Follicular regulatory T cells (Tfr)/Tfh lymphocyte proportion in spleen (left) and peripheral blood (right) of anti-PD-1-treated and isotype-treated mice (n=8 and n=4, respectively for peripheral blood and n=3 for spleen). Tfr and Tfh percentages (referred to CD4+ lymphocyte population, mean±SD) are the following: spleen, Tfr-isotype 0.49%±0.36%; Tfr-anti-PD-1 1.13%±0.53%; Tfh-isotype 2.5%±0.04%; Tfh-anti-PD-1 4.82%±2.19%; peripheral blood, Tfr-isotype 0.385%±0.22%; Tfr-anti-PD-1 3.82%±1.19%; Tfh-isotype 1.6%±1.2%; Tfh-anti-PD-1 63.55%±18.5%. Tfr population was quantified as CD25+FoxP3+ cells within CD4+CXCR5+PD-1+ cells. ns, not significant. *P<0.05; **p<0.01; ***p<0.001; ****p<0.0001.