Fig. 2a–d

Analysis of Tfh stimulated by soluble egg antigen (SEA) and soluble worm antigen (SWA). For each of three independent immunization experiments, C57BL/6 mice (five per group) were injected subcutaneously in the back with 200 μl of incomplete Freund’s adjuvant containing 50 μg of SEA, 50 μg of SWA or phosphate-buffered saline (PBS), boosted twice at 14-day intervals. Single-cell splenocyte suspensions of mice were prepared 2 weeks after the final injection. For each of three independent in vitro experiments, splenocytes of C57BL/6 normal mice were stimulated with PBS, final concentration of 10 μg/ml SEA or SWA for 72 h at 37 ℃. The splenocytes were stained with anti-CD4 FITC and anti-CXCR5 APC or isotype IgG2a control antibody for Tfh cells. a Analysis of Tfh stimulated by SEA and SWA in vivo by flow cytometry. b Comparison of Tfh stimulated by SEA and SWA in vivo. Results are expressed as mean ± SD (****P < 0.0001). d Analysis of Tfh stimulated by SEA and SWA in vitro by flow cytometry. d Comparison of Tfh stimulated by SEA and SWA in vitro (**P < 0.01). Cells were gated on the CD3⁺CD4⁺ population for analysis of Tfh cells. MDSC Myeloid-derived suppressor cells, CFSE 5(6)-carboxyfluorescein diacetate N-succinimidyl ester; for other abbreviations, see Fig. 1