Fig 8. Expansion of OT-II CD4⁺ T cells are not enhanced by systemic treatment with Indo or AG.

Mice were adoptively transferred with 1 x 10⁶ splenocytes obtained from OT-II mice and, 24 h later, immunized with 25 μg OVA protein with or without 20 μg CpG (A) or the same dose of CL097 (B) formulated in IFA. To inhibit NO, mice were given drinking water containing 2.5% AG and 1% glucose. To inhibit PGE₂, mice were injected i.p. with Indo at a dose of 2.5 mg/Kg on the day of immunization and two more injections after 4 and 8 days. Mice were sacrificed 11 days post immunization, and the number of expanded OT-II CD4⁺ T cells were quantified by cell surface staining of CD4 and CD45.1 followed by flow cytometric analysis. The total numbers of OT-II CD4⁺ T cells in spleens were calculated based on the percentages of OT-II CD4⁺ T cells and known numbers of splenocytes. Statistical differences were analyzed by one-way ANOVA between treatment groups. *p<0.05. AG: aminoguanidine hemisulfate; Indo: indomethacin.