Erformed comparing HD and TB using Mann Whitney test. For comparisons between HD, nsTB and sTB was performed using Kruskal-Wallis variance analysis followed by Dunn’s test for multiple comparisons. Analyses were performed using GraphPad Prism 5.01 software package (San Diego, CA, USA). In all cases, significance was considered at p#0.05.Results Higher frequencies of CD42CD82 (DN) ab T-cells are associated with TB severityThe proportion of CD4+, CD8+ and CD42CD82 (DN) ab Tcells, gated as described in Fig. 1A, were analyzed and compared among groups. The frequencies of CD4+ and CD8+ ab T-cells were not 22948146 different between HD and TB patients. Differences were also not observed between the frequencies of CD4+ and CD8+ ab T-cells from HD and nsTB or sTB patients, or between nsTB and sTB patients. However, the frequencies of DN ab T-cells were significantly higher in TB patients than in HD. When the comparison was done between HD and nsTB or sTB subgroups, the difference was seen between HD and sTB patients but not between HD and nsTB patients, indicating that this change happens due the severity of the 
disease. 16960-16-0 Corroborating with this finding, sTB patients present higher frequencies of DN ab T-cells than those classified as nsTB patients (Fig. 1B). The activation status of different ab T-cells subsets was analyzed based on CD69 and HLA-DR expression (Fig. 1C). The proportions of CD4+ and CD8+ ab T-cells expressing the early activation marker CD69 did not differ among the groups analyzed. However, significantly higher proportions of CD69 expressing DN ab T-cells were observed in TB patients than in HD. These differences were kept when the frequencies of CD69 expressing DN ab T-cells were compared between HD and either nsTB or sTB patients. The expression of HLA-DR was also analyzed (Fig. 1D). The 
frequencies of HLA-DR expressing CD4+, CD8+ and DN ab Tcells were significantly higher in TB patients compared with HD. Differences were also observed in the proportions of HLA-DR expressing CD4+, CD8+ and DN ab T-cells between HD and nsTB or sTB. nsTB and sTB displayed similar levels of HLA-DR expression on all ab T subsets evaluated.CD8+ cd T-cells T-cells compared with HD (Fig. 2B). The proportion of CD4+ cd T-cells from sTB patients was by itself higher than the ones observed in HD, however the same was not observed when nsTB and DH individuals were compared. Frequencies of DN cd T-cells did not differ between total TB patients and HD, but sTB patients displayed lower frequencies of this cell subset when compared with nsTB patients. Thus, lower frequencies of DN cd T-cells might suggest a severe form of AZ876 tuberculosis. Distinct of the ab T-cells, the frequencies of CD69 expressing cells were higher on CD4+, CD8+ and DN cd T-cells from TB patients compared with HD (Fig. 2C). When the CD69 expression was analyzed in CD8+ cd T-cells, its expression was 15755315 also higher in sTB patients the compared with HD. The same did not hold true for CD4+ and DN cd T-cell populations. Moreover, the opposite was seen for the DN cd T-cell subset. The increased frequencies of CD69 expressing cells in TB patients were due the high expression observed in the nsTB patients group compared to HD. The frequencies of HLA-DR expressing cells were also analyzed on CD4+, CD8+ and DN cd T-cells (Fig. 2D). The frequencies of HLA-DR expressing cells were significantly higher in TB patients compared with HD in the CD4+, CD8+ and DN cd T-cell subsets. Differences were also observed in the.Erformed comparing HD and TB using Mann Whitney test. For comparisons between HD, nsTB and sTB was performed using Kruskal-Wallis variance analysis followed by Dunn’s test for multiple comparisons. Analyses were performed using GraphPad Prism 5.01 software package (San Diego, CA, USA). In all cases, significance was considered at p#0.05.Results Higher frequencies of CD42CD82 (DN) ab T-cells are associated with TB severityThe proportion of CD4+, CD8+ and CD42CD82 (DN) ab Tcells, gated as described in Fig. 1A, were analyzed and compared among groups. The frequencies of CD4+ and CD8+ ab T-cells were not 22948146 different between HD and TB patients. Differences were also not observed between the frequencies of CD4+ and CD8+ ab T-cells from HD and nsTB or sTB patients, or between nsTB and sTB patients. However, the frequencies of DN ab T-cells were significantly higher in TB patients than in HD. When the comparison was done between HD and nsTB or sTB subgroups, the difference was seen between HD and sTB patients but not between HD and nsTB patients, indicating that this change happens due the severity of the disease. Corroborating with this finding, sTB patients present higher frequencies of DN ab T-cells than those classified as nsTB patients (Fig. 1B). The activation status of different ab T-cells subsets was analyzed based on CD69 and HLA-DR expression (Fig. 1C). The proportions of CD4+ and CD8+ ab T-cells expressing the early activation marker CD69 did not differ among the groups analyzed. However, significantly higher proportions of CD69 expressing DN ab T-cells were observed in TB patients than in HD. These differences were kept when the frequencies of CD69 expressing DN ab T-cells were compared between HD and either nsTB or sTB patients. The expression of HLA-DR was also analyzed (Fig. 1D). The frequencies of HLA-DR expressing CD4+, CD8+ and DN ab Tcells were significantly higher in TB patients compared with HD. Differences were also observed in the proportions of HLA-DR expressing CD4+, CD8+ and DN ab T-cells between HD and nsTB or sTB. nsTB and sTB displayed similar levels of HLA-DR expression on all ab T subsets evaluated.CD8+ cd T-cells T-cells compared with HD (Fig. 2B). The proportion of CD4+ cd T-cells from sTB patients was by itself higher than the ones observed in HD, however the same was not observed when nsTB and DH individuals were compared. Frequencies of DN cd T-cells did not differ between total TB patients and HD, but sTB patients displayed lower frequencies of this cell subset when compared with nsTB patients. Thus, lower frequencies of DN cd T-cells might suggest a severe form of tuberculosis. Distinct of the ab T-cells, the frequencies of CD69 expressing cells were higher on CD4+, CD8+ and DN cd T-cells from TB patients compared with HD (Fig. 2C). When the CD69 expression was analyzed in CD8+ cd T-cells, its expression was 15755315 also higher in sTB patients the compared with HD. The same did not hold true for CD4+ and DN cd T-cell populations. Moreover, the opposite was seen for the DN cd T-cell subset. The increased frequencies of CD69 expressing cells in TB patients were due the high expression observed in the nsTB patients group compared to HD. The frequencies of HLA-DR expressing cells were also analyzed on CD4+, CD8+ and DN cd T-cells (Fig. 2D). The frequencies of HLA-DR expressing cells were significantly higher in TB patients compared with HD in the CD4+, CD8+ and DN cd T-cell subsets. Differences were also observed in the.