Fig. 3: CD8α ALN-1 promotes CD8+ T cell proliferation and effector functions in response to antigen in vivo.
From: Specific targeting of IL-1β activity to CD8+ T cells allows for safe use as a vaccine adjuvant
a OT-I cells were transferred i.v. in C57BL/6 (IL-1R1−/−) mice and OVA (100 μg) was delivered i.p. 1 day later or not (PBS). OVA-immunized mice received i.p. treatments with PBS, LPS (25 μg), WT IL-1β (5 μg), CD8α ALN-1, or untargeted BcII10 ALN-1 (10 μg) every 24 h for 3 days. Flow cytometry was performed 1 day after the last treatment. b Representative flow cytometry histograms showing OT-I proliferation in C57BL/6 recipient mice. Stacked histograms summarizing OT-I proliferation in C57BL/6 (c) or C57BL/6 IL-1R1−/− (d) recipient mice. Individual stacks represent the mean percentages of proliferating OT-I cells in certain stages of cell division ± s.e.m. Shown is a pool of two independent experiments with n = 6 (c) or 10 (d) mice/group combined. e CD44/CD62L expression on CD8+ T cells (spleen) from C57BL/6 recipient mice. Bars represent the mean ± s.e.m. of a pool of two independent experiments with n = 6 mice/group combined. f Mice were treated as described in a. One week after OVA delivery, a mixed splenocyte pool (CTVhigh labeled/SIINFEKL-loaded and CTVlow labeled/nonloaded) was i.v. transferred. Flow cytometry was performed 1 day later. Representative flow cytometry histograms illustrating SIINFEKL-directed killing in C57BL/6 recipient mice (g) and quantification (h). Bars represent the mean ± s.e.m. of a pool of two independent experiments with n = 13 (OVA alone, OVA + LPS and OVA + WT IL-1β) or 10 (OVA + CD8α ALN-1 and OVA + BcII10 ALN-1) mice/group combined. i CTL responses against SIINFEKL were measured in spleens by IFN-γ ELISPOT 1 week after OVA immunization. Bars represent the mean ± s.e.m. of a pool of two independent experiments with n = 10 mice/group combined. j Representative ELISPOT pictures, depicting numbers of spots after different treatments. ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05; ns, p ≥ 0.05 by Kruskal–Wallis test with Dunn’s multiple comparisons test in c, h, and i or by unpaired Mann–Whitney U test (two-tailed) in d or by one-way ANOVA with Tukey’s multiple comparisons test in e. See also Supplementary Figs. 4 and 5.
