Fig. 3.

Mutant SOD1-expressing CD8⁺ T cells selectively trigger the death of motoneurons in vitro. (A) Motoneurons were isolated from Hb9::GFP (where GFP represents green fluorescent protein) mice and cocultured for 24, 48, 72, and 96 h with CD8⁺ T cells immunopurified from the lymph nodes (LNs) of wild-type or SOD1^G93A mice. Motoneuron survival was determined by direct counting of GFP⁺ motoneurons and expressed relative to survival in the absence of any T cells at 24 h. (B) CD8⁺ T cells were isolated either from the LNs or from the CNS of SOD1 mutant mice and cocultured with wild-type motoneurons. (C) CD8⁺ T cells were isolated from the LNs of SOD1^G93A mice at the indicated age and cocultured with wild-type motoneurons. (D) CD8⁺ T cells were isolated from SOD1^G93A mice at 150 d of age and added to motoneurons at the time of seeding (0) or 7 d later. (B–D) Survival was determined 72 h later and expressed relative to the survival in the absence of T cells. (E) Motoneurons were isolated from either wild-type or SOD1^G93A mice and cocultured with SOD1 mutant CD8⁺ T cells. Motoneuron survival was determined after 72 h and expressed relative to wild-type motoneurons cultured in the absence of T cells. The results shown are the mean values ± standard deviation (SD) of, at least, three independent experiments performed in triplicate. (B and D) Unpaired two-tailed t test, (A, C, and E) ANOVA with Tukey–Kramer’s post hoc test. *P < 0.05; **P < 0.01; ***P < 0.001; n.s, nonsignificant.