Simultaneous inhibition of COX-1/COX-2 and iNOS synergistically enhances OT-II CD4⁺ T cell proliferation stimulated with OVA protein plus CL097 in 2-week culture.

<p>5 x 10³ OT-II CD4⁺ T cells were mixed with 1 x 10⁵ congenic DCs, and incubated with a combination of OVA protein, CpG, CL097, L-NMMA (40 μM), and Indo (10 μM) for 2 weeks to measure long-term CD4⁺ T cell expansion <i>in vitro</i>. At the end of the culture period, 4 x 10⁵ splenocytes from WT C57BL/6 mice (CD45.2) were spiked into the cultures to calculate the number of OT-II CD4⁺ T cells (CD45.1). Cells were stained with anti-CD45.1 and anti-CD4 antibody to measure the expansion of OT-II CD4⁺ T cells. 7-AAD was used to exclude dead cells in the analysis. Shown are mean numbers of OT-II CD4⁺ T cells of triplicate wells ± SD. Statistical differences were analyzed by one-way ANOVA. Data are expressed as mean ± SD of triplicate wells. Data shown are representative of three independent experiments. **<i>p</i><0.001. iNOS: inducible nitric oxide synthase; COX: cyclooxygenase.