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. 2018 Jun 4;215(6):1571–1588. doi: 10.1084/jem.20171450

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© 2018 Pardi et al.

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Figure 7. — Potent Tfh cell and sustained neutralizing antibody responses are elicited by a single immunization with m1Ψ-mRNA-LNPs in nonhuman primates. (A and B) Rhesus macaques were immunized with 50 µg of CH505 Env m1Ψ-mRNA-LNPs or 100 µg of Env gp140 protein + poly-ICLC adjuvant, and Tfh cell responses in draining lymph nodes were analyzed 7 d after immunization. (A) The percentage of Tfh cells (CXCR5^hiPD1^hi) among total CD4⁺ T cells in lymph nodes is shown. (B) Lymph node cells were analyzed for Env-specific Tfh cells by activation-induced marker assay. The percentage of OX40⁺CD25⁺ cells in the Tfh gate (CD4⁺CXCR5^hiPD1^hi cells) after stimulation with Env peptide pool + protein is shown. The percentage of Env-specific Tfh cells for each sample was calculated as the percentage of OX40⁺CD25⁺ in Env-stimulated conditions minus the percentage of OX40⁺CD25⁺ in unstimulated conditions. Each point represents one animal. (C) Rhesus monkeys were immunized with 50 µg of 1086C Env m1Ψ-mRNA-LNPs at weeks 0, 4, 20, and 32, and neutralization titers (expressed as the reciprocal serum dilution resulting in ID₅₀) from preimmune and week 34 sera were determined against the MW965.26 (tier 1A) and autologous Ce1086_B2 (tier 2) viruses. Each point represents one animal. (D) Rhesus macaques were immunized with 600 µg (n = 4) or 200 µg (n = 3) of ZIKV prM-E m1Ψ-mRNA-LNPs, and the antibody response was measured by PRNT against ZIKV MR-766. Points represent individual monkeys; horizontal lines indicate the mean. Statistical analysis: (A–B) two-way ANOVA with Bonferroni correction, *, P < 0.05; (D) dose groups were compared by Kruskal-Wallis test, *, P > 0.05 for all comparisons.