Regulatory Role of B Cells and Its Subsets in Hepatitis E Virus Infection

Figure 8

Flow cytometric analysis showing the effect of depletion of CD19⁺IL-10⁺ cells on HEV-rORF2p specific CD4 and CD8 T cells response in terms of IFN-γ expression. B cells were magnetically sorted from PBMCs from 33 acute hepatitis E patients, 22 hepatitis E recovered individuals and 52 healthy controls, and were stimulated with/without HEV-rORF2p followed by detection and isolation of IL-10 expressing B cells. Sorted CD19⁺IL-10⁺ and CD19^-IL-10^- cells were co-cultured with PBMCs derived from the same patients/controls and were intracellularly stained to determine the expression of IFN-γ on CD4⁺ T cells (%CD3⁺CD4⁺IFN- γ⁺) and CD8⁺ T cells (%CD3⁺CD8⁺IFN- γ⁺). Expression of IFN-γ on HEV-rORF2p specific CD4 T cells (CD3⁺CD4⁺IFN-γ⁺) in (a) acute hepatitis E patients, (b) hepatitis E recovered individuals, (c) healthy controls and CD8 T cells (CD3⁺CD8⁺IFN-γ⁺) in (d) acute hepatitis E patients, (e) hepatitis E recovered individuals, (f) healthy controls were assessed pre vs. post CD19⁺IL-10⁺ cells depletion. Pair wise comparisons were done on matched samples wherein dots represent individual values.