Fig. 4: Lmna^−/− CD4⁺ T-cells show impaired in vivo and in vitro activation in response to VACV infection

a, b WT CD45.1⁺/CD45.2⁺ mice were adoptively transferred with a mix of CD4⁺ T-cells from CD45.1⁺/OTII WT and CD45.2⁺/OTII Lmna^−/− mice and then intradermally infected in the footpad with VACV-OVA for 2 days. Spleen and popliteal lymph nodes were analyzed, and the expression of a CD25 and b CD69 was quantified (n = 6 mice). Data are means±SEM analyzed by unpaired Student’s t-test c Splenocytes from CD45.1⁺ WT/OTII and CD45.2⁺ Lmna^−/−/OTII mice were mixed, stained with cell violet, and cultured in the presence of OVA for 2 days. CD4⁺ T-cell proliferation was quantified and the number of cell divisions is shown in the graph. Data are means±SEM of six independent experiments analyzed by two-way ANOVA with Bonferroni’s multiple comparison test. d, e CD4⁺ T-cells from CD45.1⁺ WT and CD45.2⁺ Lmna^−/− mice were mixed 1:1 and adoptively transferred to CD45.1/CD45.2 WT mice. Recipient mice were then intraperitoneally infected with VACV for 7 days. The peritoneal exudates and mesenteric lymph nodes were analyzed, and the percentage of d Ki67⁺ or e PCNA⁺ CD4⁺ T-cells was quantified (n = 5 mice from 2 independent experiments) Data are means±SEM analyzed by unpaired Student’s t-test. f CD45.1⁺ WT recipient mice were transplanted with a 1:1 mix CD45.1⁺ WT and CD45.2⁺ Lmna^−/− bone marrow and infected intraperitoneally with VACV. Mice were injected with BrdU, and 24 h later the percentage of BrdU⁺ CD4⁺ T-cells was quantified in mesenteric lymph nodes, peritoneal exudate, and spleen (n = 3 mice from 1 representative experiment) Data are means±SEM analyzed by paired Student’s t-test. g CD4⁺ T-cells from CD45.2⁺ WT or CD45.2⁺ Lmna^−/− mice were mixed 1:1 and adoptively transferred to CD45.1⁺/CD45.2⁺ WT mice. Recipient mice were infected intraperitoneally with VACV for 6 days and injected with BrdU. After 24 h, the percentage of BrdU⁺ CD4⁺ T-cells was quantified in the peritoneal exudate and mesenteric lymph nodes (n = 5 and 5 mice). Data are means±SEM analyzed by paired Student’s t-test. ***P < 0.001