Figure 6

Clone 10 treatment reduces T cell neurotoxicity. Peripheral blood mononuclear cells (PBMCs) incubated in the absence (non-act.) or presence of anti-CD3 and anti-CD28 (act.) were applied to human fetal neurons, and neuronal killing was then assessed (A). Using ImageXpress analysis, the number of microtubule-associated protein 2 (MAP2) + neurons in defined fields per well was enumerated; neuronal counts were reduced by activated T cells, representing neuronal death, and this was prevented by clone 10 treatment of T cells (B). Values are mean ± SEM of quadruplicate wells; ***P <0.001 (one-way analysis of variance (ANOVA) with Tukey’s post hoc comparisons). (C) Representative images of MAP2-labeled neurons. Fluorescence-activated cell sorting (FACS) analysis was used to estimate Granzyme B levels in CD3+ T cells with or without treatment with clone 10 (D). Values in (D) are mean ± SEM of triplicates samples; ***P <0.001 (one-way ANOVA with Tukey’s post hoc comparisons).