Figure 4
Flow cytometric analysis of GC B-cell and TFH-cell formation in Lungs. TLN+MLN and ILN were analyzed 10 days after s.c./i.n., i.n./i.n. and s.c./s.c. immunization with ScpA/CAF01 compared with naïve background levels. Mice were intravascular stained by iv injection of FITC conjugated anti-CD45, 3 min before being killed. (a) Dot plots (concatenated within groups) showing GL7 vs. CD95 expression on B220+CD19+ B cells from lungs (gated for iv.CD45−) or lymph nodes (LNs; gated for IgD−). (b) Boxplots (min/max; median w. s.e.m.) showing total numbers of GC B cells in lungs (top), TLN+MLN (middle) and ILN (bottom). (c) Dot plot showing PD1 vs. CXCR5 expression on CD44+CD4+ T cells from lungs (gated for iv.CD45−) and lymph nodes. (d) Boxplots (min/max; median w. s.e.m.) showing total numbers of TFH cells in lungs (top), TLN+MLN (middle), and ILN (bottom). Statistical analysis: one-way ANOVA using Tukey’s post-test *P<0.05. ANOVA, analysis of variance; FITC, fluorescein isothiocyanate; GC, germinal center; ILN, inguinal lymph node; i.n., intranasal; iv, intravenously; MLN, mediastinal lymph node; s.c., subcutaneous; ScpA, streptococcal C5a peptidase; TFH, follicular T helper cells; TLN, tracheobronchial lymph nodes.
