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. 2022 May 13;119(20):e2118312119. doi: 10.1073/pnas.2118312119

Fig. 2.

Fig. 2.

Spike-specific mucosal and systemic effector CD8 T cell responses to adjuvanted vaccines. C57BL/6 mice were IN or SQ vaccinated twice (21 d apart) with recombinant SARS-CoV-2 coronavirus S protein formulated in ADJ (5%) and CpG (5 μg) or GLA (5 μg). To identify circulating/vascular cells in the lungs, mice were injected intravenously with labeled anti-CD45.2 antibodies, 3 min prior to killing (CD45.2−, nonvascular; CD45.2+, vascular). At day 8 postimmunization, single-cell suspensions from lungs and spleens were stained with viability dye, followed by H2-Kb/S525 tetramers in combination with anti-CD4, CD8, CD44, CD69, CD103, CD49a, CX3CR1, KLRG1, CD127, CXCR3, and CD62L antibodies. (A and B) FACS plots show numbers and percentages of S525 tetramer-binding cells among CD8 T cells from IN-vaccinated mice (A) and SQ-vaccinated mice (B). (C) Relative proportions of vascular (CD45.2+) and nonvascular (CD45.2−) cells among S525-specific CD8 T cells. (D–I) Percentages of CD103, CD69, CD49a, CX3CR1, KLRG1, CD127, CXCR3, and CD62L+ cells among S525-specific CD8 T cells in lungs and spleen. Data represent one of two independent experiments. Asterisks indicate significance at *P < 0.05, **P < 0.005, ***P < 0.0005, and ****P < 0.00005. Data in each graph indicate mean ± SEM.