Data for: DosR proteins of Mycobacterium tuberculosis upregulate effector T cells and down regulate T regulatory cells in TB patients and their healthy contacts

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.

补充图S1。重组蛋白Rv2627与Rv2628的克隆与表达

a) 基因Rv2627（泳道L1，产物大小1242bp）与基因Rv2628（泳道L2，产物大小363bp）的阳性质粒PCR扩增结果，M为分子量Marker。
b) 天然条件下纯化的Rv2627与Rv2628蛋白的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE, Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis）结果：M为分子量Marker，L1为纯化的Rv2627蛋白，L2为纯化的Rv2628蛋白；Rv2627的分子量为46.5 kDa，Rv2628的分子量为13.1 kDa。
c) 纯化的Rv2627与Rv2628蛋白的蛋白质免疫印迹（Western Blot）结果：M为分子量Marker，L1为纯化的Rv2627蛋白，L2为纯化的Rv2628蛋白。

补充图S2。利用标记的未刺激与刺激细胞及FMO对照(Fluorescence Minus One)获取并分析CD3阳性T细胞的设门策略：
a) CD4+IFN-γ+双阳性细胞；b) CD4+CD45RA+双阳性细胞；c) CD4+CD45RO+双阳性细胞；d) CD8+IFN-γ+双阳性细胞；e) CD8+CD45RA+双阳性细胞；f) CD8+CD45RO+双阳性细胞；g) 调节性T细胞（Treg, Regulatory T cells，CD4+CD25+FoxP3+）。

补充图S3。正常人群（n=20）、接触者（n=20）与肺结核（PTB, Pulmonary Tuberculosis）患者（n=20）三类样本群体中不同T细胞标志物的基础占比。
相较于接触者与肺结核患者，正常人群的CD4+CD45RA+、CD4+CD45RO+、CD8+CD45RA+、CD8+CD45RO+阳性T细胞占比最高；而肺结核患者的调节性T细胞占比最高。样本间的统计学差异采用非配对样本的曼-惠特尼U检验（Mann-Whitney U test）计算。结果以均值±标准差（mean ± SD）表示，*P < 0.05，**P < 0.01，***P < 0.001，****P < 0.0001。

补充图S4。正常人群（n=15）、接触者（n=15）与肺结核患者（n=15）三类样本群体的外周血单个核细胞（PBMC, Peripheral Blood Mononuclear Cells）基础细胞因子分泌谱。
接触者的干扰素γ（IFN-γ, Interferon Gamma）、白细胞介素2（IL-2, Interleukin 2）等细胞因子基础水平更高，而肺结核患者的白细胞介素4（IL-4, Interleukin 4）、转化生长因子β（TGF-β, Transforming Growth Factor Beta）基础水平更高。样本间的统计学差异采用非配对样本的曼-惠特尼U检验计算。结果以均值±标准差表示，*P < 0.05，**P < 0.01，***P < 0.001。

补充图S5。Rv2627与Rv2628通过提高干扰素γ的分泌量，使接触者与肺结核患者的免疫反应向Th1型（辅助性T细胞1型，T helper cell 1）倾斜。
在接触者群体中，仅Rv2628可显著提高IFN-γ/IL-4比值；而在肺结核患者样本中，两种蛋白均可提高该比值。未刺激与刺激样本间的统计学差异采用配对样本的威尔科克森秩和检验（Wilcoxon rank sum test）计算。结果以均值±标准差表示，*P < 0.05。