Sex-Specific Immune Responses to Seasonal Influenza Vaccination in Diabetic Individuals: Implications for Vaccine Efficacy

Figure 4

Determining population percentage of memory B cells, immature/transitional B cells, activated B cells, and plasma cells. (a) B220+ cells and B220^low populations are gated out from live single splenocyte suspension keeping the unstained sample as a reference for gating as usual. CD19+ cells are gated out from B220+ cells and CD19^low/neg cell populations are gated out from B220^low populations. (b) On the B220+CD19+ cell populations, CD27+MHCII+ populations are gated, on which CD40+CD80+ populations are determined to get B220/CD45R+CD19+CD27+MHCII+CD40+CD80+ memory B cells. (c) IgM^high+IgD^low/neg population (yellow box) and IgM+IgD+ (thick black box) population are gated on B220+CD19+ populations. B220/CD45R+CD19+IgM^high+IgD^low/neg+CD43− immature/transitional B cells and B220/CD45R+CD19+IgM+IgD+MHCII+CD138− activated B cell population are determined. (d) From the B220^low CD19^low/neg populations IgM−IgD− cells are determined to obtain B220^low CD19^low/neg IgM−IgD−CD138+ plasma cells. (e) The bar diagram represents the memory B cells and immature/transitional B cells. (f) The bar diagram represents the activated B cells and plasma cells. The percentage exhibited in the Y axis of (e) and (f) is the percentage within the total 10,000 splenocytes analyzed. Hence, for comparison for absolute cell number, the conversion: 1% in the Y axis = 100 cells. D−, nondiabetic groups; D+, diabetic groups. Data in graphs are the representative images derived from at least four independent experiments (, , and ). For each group n = 5, the error bar indicates the standard deviation.