Study of T-cell functional avidity and in vivo T-cell specific killing activity.

<p>Pooled splenocytes from control and IL-12₅₀+CTB groups (three to six mice/group), were analyzed for functional avidity of the V3- loop specific T-cells at (A) 10 and (B) 30 days after immunization, by ELISPOT using serial dilutions of the peptide from 20 to 0.00002 µg/ml for triplicate wells. Data represents the average percentage of the maximal response. SD50: sensitizing dose of peptide required to yield 50% of maximal T-cell triggering IFN-γ production. (C) <i>In vivo</i> cytotoxicity was analyzed adoptive transfer of control CFSElow and Env-peptide pulsed CFSEhigh dye cells. At 10 and 30 days after immunization, individual spleens of mice from naïve (I), DNA-EnvB/MVA-EnvB (II) and DNA-EnvB+DNA-IL-12₅₀+CTB/MVA-EnvB+CTB (III) groups were analyzed at the indicated times after adoptive transfer. A representative histogram is depicted in the left panel. Dots represent each analyzed mice. Lines represent median Statistical differences between groups: *p<0.05; **p<0.01 by Mann-Whitney test.