Data for: DosR proteins of Mycobacterium tuberculosis upregulate effector T cells and down regulate T regulatory cells in TB patients and their healthy contacts
Description
Fig S1. Cloning and Expression of recombinant proteins Rv2627 and Rv2628 a) Positive plasmid PCR of gene Rv2627 (L1) with product size 1242bp and gene Rv2628 (L2) with product size 363bp and M- Marker. b) SDS PAGE of purified Rv2627 and Rv2628 protein in native condition. M-Marker and L1- Rv2627 and L2- Rv2628 purified protein. The molecular weight of Rv2627 was 46.5 kDa and Rv2628 was 13.1 kDa. c) Western blot of purified Rv2627 and Rv2628. M-Marker and L1- Rv2627 and L2- Rv2628 purified protein. Fig S2. Gating strategies used for acquiring and analysis of CD3+ cells using labeled un-stimulated and stimulated cells and their FMO controls a) CD4+ IFN-γ+, b) CD4+CD45RA+, c) CD4+CD45RO+, d) CD8+IFN-γ+, e) CD8+CD45RA+, f) CD8+ CD45RO+ and g) Treg (CD4+CD25+FoxP3+). Fig S3. Basal percentage of different T cell markers in the three sample population of Normal (n=20), Contacts (n=20) and PTB patients (n=20). The normal individuals had highest percentage of CD4+CD45RA+, CD4+CD45RO+, CD8+CD45RA+, CD8+CD45RO+ cells as compared to that of contacts and patients whereas the PTB patients had highest percentage of Treg cells. Statistical differences between samples were calculated by the Mann-Whitney U test for unpaired samples. Values were shown as mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001. Fig S4. Basal Cytokine production profile in PBMC of three sample population of Normal (n=15), Contacts (n=15) and PTB patients (n=15). The contacts had higher basal level of cytokines like IFN-γ and IL-2 whereas PTB patients had higher basal levels of cytokines like IL-4 and TGF-. Statistical differences between samples were calculated by the Mann-Whitney U test for unpaired samples. Values were shown as mean ± SD, *P < 0.05, **P < 0.01 and ***P < 0.001. Fig S5. Rv2627 and Rv2628 skew the immune response towards Th1 in both contacts and patients by increasing the IFN-γ production In contacts, IFN-γ/IL-4 ratio increased significantly only for Rv2628, whereas in PTB samples the ratio increased for both the proteins. Statistical differences between unstimulated and stimulated samples were calculated by the Wilcoxon rank sum test for paired samples. Values were shown as mean ± SD, *P < 0.05.