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Fig 3 | Journal of Neuroinflammation

Fig 3

From: CXCR5-negative natural killer cells ameliorate experimental autoimmune myasthenia gravis by suppressing follicular helper T cells

Fig 3

NK cells inhibit CD4+ T cells and Tfh cells in vitro. a Rat splenocytes were co-cultured with NK cells or not. Representative flow cytometric plots of Tfh cells (CD4+CXCR5+ICOS+) among CD4+ T cells (left). Frequencies of Tfh cells in CD4+ T cells were downregulated by NK cells. b, c Mouse splenocytes were co-cultured with NK cells or not. CD4+ T cell and Tfh (CD4+CXCR5+PD1+) cell numbers were counted by flow cytometry and shown in relative forms (b, left and median). The Tfh cell frequencies in CD4+ T cells were not changed (b, right). The apoptosis of CD4+ T cells was determined by staining cells with CD4 and Annexin V (c). d, e Splenic CD3+ T were co-cultured with NK cells or not. The CD4+ T cells and Tfh cells were determined by flow cytometry (d). The apoptosis of CD4+ T cells and Tfh cells was determined by staining cells with Annexin V (e). f, g Mouse splenocytes were cultured with IL-15 or not. IL-15 increased NK cells dramatically in vitro (f, left). IL-15 treatment decreased CD4+ T cell numbers and Tfh cell frequencies in CD4+ T cells (f, median and right) and induced apoptosis of CD4+ T cells (g). h Mouse splenocytes were labeled with CFSE and cultured with IL-15 or not. Cells were then stained with CD19 to detect the proliferation of B cells. Representative flow cytometric histogram of CFSE dilutions of B cells. Data were from one (a, h) or two independent experiments (bg). Data were presented as mean ± SEM. Unpaired Student’s t test was used. ns means not significant, *p < 0.05, **p < 0.01, ***p < 0.001

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