Percentage peripheral blood in three groups of individuals

Percentage of CD4+ T cells and its regulatory subsets
(Tregs) within CD4+ cells and frequencies of cells expressing PD-1
and PD-L1 in peripheral blood in three groups of individuals are shown in Table
2.

CD+CD25+ T-cells were gated and frequencies
were analysed as a percentage of CD4+cells. CD4+CD25+Foxp3+
T-cells (Tregs)  frequency were gated and
analysed as a percentage of CD4+CD25+ cells. The patients
with active pulmonary tuberculosis had an average proportion of 29.51%±11.93 CD4+ lymphocytes ,
similar to Latent tuberculosis (29.23%±8.16)and house hold
contacts(28.16%±9.73), p=0.8420. (Fig. 1A)

Within the CD4+ lymphocytes a significantly higher
percentages of CD4+CD25+ T-cells, CD4+CD25+Foxp3+
T-cells and CD4+CD25+Foxp3+PD1+ T-cells were observed in
APTBs than that in HCs ( 19.05% vs 4.58%, p<0.0001; 11.3% vs 1.84%, p<0.0001 and 14.54% vs 0.37%, p<0.0001 respectively). Similar trend was also observed from APTBs to  LTBIs (19.05% vs 6.45%, p<0.0001; 11.30% vs 4.32%, p<0.0001 and 14.54% vs 2.99% p<0.0001 respectively) and from  LTBIs to HCs (6.45% vs 4.58%,p=0.001; 4.32% vs 1.84% , p<0.0001 and 2.99% vs 0.37%, p<0.0001 respectively). A significantly lower percentage of CD4+CD25-Foxp3- was observed in APTBs than that in HCs (47.47% vs 82.69%, p<0.0001). A significantly negative correlation between CD4+CD25-Foxp3- and CD4+CD25+Foxp3+ (p<0.0001, R2=0.4248) in the study cohort was observed. (Fig. 1B, 1C, 1D, 1E)  The percentage of PD-1 and PD-L1 expressing CD4+ T- cells in APTBs patients was significantly higher than in HCs (41.93% vs 27.95%, p=0.0004 and 7.13% vs 0.80%, p<0.0001 respectively, Fig 2A,2B). Because PD-1 mediated inhibition of effector T-cells function required engagement with the expression of its ligands on Antigen presenting cells (APC), we further examined the expression of PD1 ligands (PD-L1) on CD14+ monocytes. Significantly greater increases of PD-L1 on CD14+ cells were observed in APTBs compared to  HCs(6.78% vs 0.90%, p<0.0001, Fig. 2D). We further examined the expression level  of PD1 both in CD4+CD25+Foxp3+  T- cells and CD4+CD25+Foxp3-  T- cells and Significantly high expression level of PD1 (MFI) in CD4+CD25+ Foxp3+ Tregs than in CD4+CD25+ Foxp3- Tregs (111.5 vs 76.03, p<0.0001, Fig. 2E) was observed (Fig 5B) . PD- L1 expression in CD4+CD25+Foxp3+ cells   could not be evaluated due to technical resion.

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