Previous studies on the effects of IL-2 on IL-17 secretion in non-HIV models have shown divergent results

Previous studies on the effects of IL-2 on IL-17 secretion in non-HIV models have shown divergent results. frequency of TH17 cells declined, while counts of TH17 cells did not change due FBL1 to an expansion of the CD4+ na?ve T cell population (CD27+CD45RA+). Counts of HIV-1 Gag-specific T cells declined modestly, but CMV pp65 and CD3/CD28 stimulated populations did not change. Hence, in contrast with recent studies, our results suggest IL-2 is not a potent in vivo regulator of TH17 cell populations in HIV-1 disease. However, IL-2-mediated T-reg expansions may selectively reduce responses to certain antigen-specific populations, such as HIV-1 Gag. Electronic supplementary material The online version of this article (doi:10.1007/s10875-010-9432-3) contains supplementary material, which is available to authorized users. (SEB) at a concentration of 5?g/ml (Sigma Aldrich). Soluble anti-CD3 (0.5?g/mL, clone HIT3; BD Biosciences) and soluble anti-CD28 (0.5?g/mL clone 28.2; BD Biosciences) was used as previously described in the ELISPOT assay [4]. Spot totals for duplicate wells were averaged, and all spot numbers were normalized to numbers of IFN- spot-forming models (SFU) per 1??105 PBMCs. Spot values from medium control wells were subtracted to determine responses to each peptide. Responses 100 spots/105 PBMCs were considered as a positive responding populace. Statistical Analysis We tested for IL-2-associated changes on test variables by applying the sign rank test to test if the difference between pre- and post-IL-2 administration time periods within each group differed from zero. We compared differences in measurements between groups at baseline and during the post-IL-2 time period with the Wilcoxon two-sample test. We assessed correlations between continuous variables by use of the Spearman rank correlation test. Data were manipulated and statistical assessments performed in the SAS System 9.2 for Windows XP. We employed GraphPad/Prism (La Jolla, CA, USA) to display results from this study. Results Study Populace and Description of HAART and IL-2 Therapy Using HIV-1 as a model to study the effects of IL-2 on IL-17 production in Chlorquinaldol vivo, we studied 18 participants from a randomized clinical trial of IL-2 performed at the University of California San Francisco. We restricted selection of IL-2 treated subjects to those who completed at least five cycles (out of a possible six) of IL-2 (Table?I). All 18 adults remained on ART for at least 1?12 months after randomization. Of these 18 subjects, 11 subjects received IL-2 and are referred to as ART + IL-2. The remaining seven comparison subjects who received ART therapy only during the study period are referred to as ART. Virologic responses to ART were excellent among all participants who achieved and maintained complete virologic suppression for the duration of the study. We did not observe a viral rebound effect among those who either did or did not receive IL-2. We focused on steps at two time points. Visit 1 was designated at the time when a viral load of less than 500 copies/mL had been achieved on ART, but before IL-2 had been administered in the ART + IL-2 group, and as corresponding time in the ART only group. Visit 2 was approximately 48?weeks later, and represented a time when at least five cycles of IL-2 therapy had been administered in Chlorquinaldol the ART + IL-2, and represented a corresponding time on treatment in the ART alone group. Table?I Demographic, Clinical, and Laboratory Measures at Study Entry All Patients on Suppressive ART, but Prior to Receipt of Study Drug = b indicated the gated population subsequently analyzed. Fluorescence minus one (FMO) samples were used to define the gates used Baseline T Cell Responses to Stimuli at Visit 1 (Prior to IL-2 on Both Randomized Populations) We observed that this magnitude of T cell response cell counts at visit 1 (pre IL-2 trial period) in response to CD3/CD28, HIV-1 Gag, and CMV pp65 peptide pools in the two patient populations did not differ. Thus, the patient groups intended to subsequently receive IL-2 and not receive IL-2 did not differ from one another at baseline and, hence, show evidence the groups were well randomized (Table?I). We observed that IL-17 responses were not induced by viral peptide pool stimulation of T cells in either CD4+ or CD8+ T cells, but Chlorquinaldol were induced by TCR (anti-CD3/CD28) cross-linking in both groups by CD4+ T cells and did not differ between the two groups (Fig.?2). Open in a separate windows Fig.?2 Counts of.