Figure 6
Figure 6. LT signaling during LCMV-induced PLN remodeling. (A) Experimental layout for LT blocking in WT mice and B-cell reconstitution in JHT mice, followed by LCMV infection. WT mice received 2 injections on day 0 and 3 after infection of 100 μg of control or LTβR-Ig. JHT mice were reconstituted with WT or LTβ−/− B cells as described in Figure 4A. (B) Representative OPT images of inguinal LN isolated from day 8 (D8) LCMV-infected WT, WT + LTβR-Ig, JHT + WT B cells, and JHT + LTβ−/− B cells mice. Scale bar = 400 μm. (C) Inguinal LN volume on day 0 (D0) or day 8 (D8) after infection after treatment of mice with control Ig or LTβR-Ig or in JHT mice reconstituted with WT or LTβ−/− B cells. (D) Total HEV length on day 0 (D0) or day 8 (D8) after infection as in panel C. All day 8 values in panels C and D were significantly greater than on corresponding day 0 (Student t test). *P < .05, **P < .01, ***P < .001 comparing day 8 LCMV “WT + control Ig” vs “WT + LTβR-Ig” and “JHT + WT B cells” vs “JHT + LTβ−/− B cells,” respectively. No significant difference was found in panels C and D between day 8 “WT + control Ig” and “JHT + WT B cells” nor between “WT + LTβR-Ig” and “JHT + LTβ−/− B cells” (1-way ANOVA). (E) Percentage of B-cell volume over total PLN volume in WT mice on day 0 (D0) or day 8 (D8) after infection after treatment of mice with control Ig or LTβR-Ig. ***P < .001. (F) Absolute B-cell number per inguinal LN in mice treated with control Ig or LTβR-Ig on day 8 after infection. n.s. indicates not significant. Samples were pooled from 3 to 9 PLNs from 2 to 8 mice in 2 independent experiments. Data are shown as mean ± SEM. (G) Average HEV segment length on day 0, 2, 5, and 8 after infection. ▭ indicate 20 to 100 μm; , 100 to 200 μm; ▬, greater than 200 μm. On day 2, the proportion of segments less than 100 μm is significantly increased, whereas the proportion of segments greater than 200 μm is increased on day 8 after infection. Samples were pooled from a total of 4 to 5 inguinal LNs from 2 to 3 mice for each time point. (H) Total HEV segment number in WT and JHT mice infected with LCMV. Filled columns represent day 8 after infection. (I) Percentage of HEV with diameter greater than 50 μm during LCMV infection. Filled columns represent day 8 after infection. Data are shown as mean ± SEM. *P < .05, ***P < .001 compared with “D0” (1-way ANOVA).

LT signaling during LCMV-induced PLN remodeling. (A) Experimental layout for LT blocking in WT mice and B-cell reconstitution in JHT mice, followed by LCMV infection. WT mice received 2 injections on day 0 and 3 after infection of 100 μg of control or LTβR-Ig. JHT mice were reconstituted with WT or LTβ−/− B cells as described in Figure 4A. (B) Representative OPT images of inguinal LN isolated from day 8 (D8) LCMV-infected WT, WT + LTβR-Ig, JHT + WT B cells, and JHT + LTβ−/− B cells mice. Scale bar = 400 μm. (C) Inguinal LN volume on day 0 (D0) or day 8 (D8) after infection after treatment of mice with control Ig or LTβR-Ig or in JHT mice reconstituted with WT or LTβ−/− B cells. (D) Total HEV length on day 0 (D0) or day 8 (D8) after infection as in panel C. All day 8 values in panels C and D were significantly greater than on corresponding day 0 (Student t test). *P < .05, **P < .01, ***P < .001 comparing day 8 LCMV “WT + control Ig” vs “WT + LTβR-Ig” and “JHT + WT B cells” vs “JHT + LTβ−/− B cells,” respectively. No significant difference was found in panels C and D between day 8 “WT + control Ig” and “JHT + WT B cells” nor between “WT + LTβR-Ig” and “JHT + LTβ−/− B cells” (1-way ANOVA). (E) Percentage of B-cell volume over total PLN volume in WT mice on day 0 (D0) or day 8 (D8) after infection after treatment of mice with control Ig or LTβR-Ig. ***P < .001. (F) Absolute B-cell number per inguinal LN in mice treated with control Ig or LTβR-Ig on day 8 after infection. n.s. indicates not significant. Samples were pooled from 3 to 9 PLNs from 2 to 8 mice in 2 independent experiments. Data are shown as mean ± SEM. (G) Average HEV segment length on day 0, 2, 5, and 8 after infection. ▭ indicate 20 to 100 μm; , 100 to 200 μm; ▬, greater than 200 μm. On day 2, the proportion of segments less than 100 μm is significantly increased, whereas the proportion of segments greater than 200 μm is increased on day 8 after infection. Samples were pooled from a total of 4 to 5 inguinal LNs from 2 to 3 mice for each time point. (H) Total HEV segment number in WT and JHT mice infected with LCMV. Filled columns represent day 8 after infection. (I) Percentage of HEV with diameter greater than 50 μm during LCMV infection. Filled columns represent day 8 after infection. Data are shown as mean ± SEM. *P < .05, ***P < .001 compared with “D0” (1-way ANOVA).

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