Frequency of HLA-DR⁺ Teff cells is elevated in PTB.

PBMC were stained with Avid, anti-CD3, anti-CD4, anti-CD45RA, anti-CD127, anti-CD25 and anti-HLA-DR, anti-CD38 and anti-PD-1. Stained samples were acquired on a FACS Aria Fusion after using appropriate single color compensation controls. A sequential gating strategy was employed to arrive at live CD3+CD4+CD45RA-CD127hiCD25lo Teff cells. Expression of HLA-DR, CD38 and PD-1 was studied in the Teff fraction. Representative HLA-DR staining on Teff cells from different clinical categories is shown (A). Frequency of HLA-DR+ Teff cells from different clinical categories was plotted (B). SPICE analyses of Boolean gating data of HLA-DR, CD38 and PD-1 expression on Teff cells derived from FlowJo was carried out and frequencies of Teff cells expressing different combinations of markers is shown (C). Data shown is median frequency with range from multiple donors (IGRA-ve N = 9, IGRA+ve N = 11, PTB N = 27, ATT 6 months N = 8) in each clinical category. P value for (B) was determined by non-parametric One-Way ANOVA Kruskal–Wallis test and Dunn’s multiple comparisons test. P value for (C) was determined by Mann Whitney test. ***p < 0.001, **p < 0.01, *p < 0.05.