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. 2020 Nov 21;53(6):1281–1295.e5. doi: 10.1016/j.immuni.2020.11.009

Figure 6.

Figure 6

SARS-CoV-2 mRNA Vaccines Drive High Expression of Tfh Signature Molecules

Mice were i.m. immunized with SARS-CoV-2 mRNA vaccines or rRBD-AddaVax. For (A)–(D), (G), (H), and (J), inguinal LNs and pooled inguinal and popliteal LNs ([E], [I], and [K]) were analyzed. In (A)–(E) and (G)–(K), data were analyzed at 7 days and in (F) at 60 days post-immunization.

(A) Representative analysis of IFN-γ+ and IL-4+ Tfh cells (PD-1+CXCR5+) measured by ICS after PMA/ionomycin stimulation. Cells were pre-gated on live CD4+B220−CD44hi populations.

(B) Frequency of IFN-γ+ Tfh cells as described in (A). In (B), (C), and (H), dotted line represents an average of 35 unstimulated samples.

(C) Frequency of IL-4+ Tfh cells as described in (A).

(D and E) Ratio of IL-4+/IFN-γ+ Tfh cells measured after (D) PMA/ionomycin stimulation or (E) SARS-CoV-2 peptide stimulation. Data used for (E) are detailed in Figure S6.

(F) Ratio of IgG1/IgG2a (left) and IgG1/IgG2b titers (right). Data used for this ratio is detailed in Figure 4.

(G) Representative analysis of IL-21+ Tfh cells (PD-1+CXCR5+) measured by ICS after PMA/ionomycin stimulation. Cells were pre-gated on live CD4+B220−CD44hi populations.

(H) Frequency of IL-21+ Tfh cells as described in (G).

(I) Relative expression of Ascl-2 on purified naive CD4 and Tfh cells (PD-1+CXCR5+) measured by qPCR.

(J and K) Mean fluorescence intensity (MFI) of (J) CXCR5 or (K) ICOS on Tfh cells (CXCR5+Bcl-6+).

In (A)–(D) (F), (G), (H), and (J), n = 9 mice per group were analyzed. Data were combined from three independent experiments. In (I) and (K), n = 8 mice per group were analyzed. Data were combined from two independent experiments. In (B)–(E) and (H)–(K), mean ± SEM is shown. In (F), ratios of Ab titers are shown as geometric mean ± geometric SD. Each data point represents an individual mouse. One-way-ANOVA with Bonferroni correction or unpaired two-tailed Mann-Whitney U tests were conducted according to the distribution of the data. ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, ∗∗∗∗p ≤ 0.0001 See also Figure S6 and Table S1.