Fig. 5.

Tfh frequencies are increased by adjuvants associated with protection from challenge.

C57BL/6 mice were vaccinated a single time IM with VLP, with or without adjuvant. Seven days after vaccination, the draining popliteal LN was isolated. (a) Untouched T cells were isolated from the LN (n = 3 or 4/group). RNA was isolated from the purified T cells and pooled for each vaccination group. Samples were then evaluated using the SABiosciences T and B cell activation PCR array in triplicate. For the volcano plot, the fold difference is the average of triplicate, and the p value was calculated comparing animals vaccinated with VLP to those vaccinated with VLP and polyICLC. (b) Seven days after vaccination, cells from the draining LN were evaluated for Tfh populations. Cells were gated on viable lymphocytes after doublet exclusion, and then on CD4 + T cells expressing CXCR5 and PD1. Median and IQR shown. (c) CD3 + CD4 + CXCR5 + PD1 + cell population in mice vaccinated with VLP and polyICLC. ICOS and Bcl6 expression of this population is shown. Data shown are pooled from three separate vaccination experiments. Comparisons between animals receiving VLP with or with adjuvant are shown, where “*” indicates 0.01 < p < 0.05 (Kruskal–Wallis test).