Analysis of FOXP3+ Regulatory T Cells That Display Apparent Viral Antigen Specificity during Chronic Hepatitis C Virus Infection
Figure 4
CD25+/↑ cells are suppressive.
(A) A simple illustration of the CD4 gate. The co-cultured cells from the patient were sorted as described in Figure 2A, except that a CD4 gate was introduced in addition to the CD3 gate, into P5 (CFSE+CD25+), P6 (CFSE+CD25−) and P9 (CD4+CD3+CFSE−) fractions. (B) The in vitro suppression assay. The target cells were a HCV-specific CD8 T cell line (comprised of autologous CD8 T cells and monocytes, cultured for 5 days in the presence of HCVpp). 1×105 of the sorted cells (P5, P6 or P9) were mixed with 2×105 target cells and 2×104 feeders (autologous iDC). This co-culture was stimulated with HCVpp for 7 days, and analysed for Ki67 expression by flow cyometry, gating on CD8+ target cells. (Representative data of N = 3 independent experiments using different donors).