Analysis of transcription factors, microRNAs and cytokines involved in T lymphocyte differentiation in patients with tuberculosis after directly observed treatment short-course
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Tuberculosis (Tb) is an infectious disease in which the immune system plays an important role. MicroRNAs are involved in the development and maintenance of CD4 T lymphocyte subpopulations. miR-326 regulates the differentiation to Th17 cells and miR-29 correlates with the Th1 response. The aim of this study was to determine the role of microRNAs, Transcription Factors, and cytokines in Th differentiation before and after the directly observed treatment short-course (DOTS). Peripheral blood mononuclear cells and serum from Tb patients were collected at times 0 (before therapy), 2 (after the intensive phase), and 6 months (after the holding phase). The cells were cultivated in presence or absence of ESAT-6 (10 μg/ml) and CFP-10 (10 μg/ml). Transcription Factor and microRNA expressions were analyzed by qPCR and cytokine production in both serum and culture supernatant using ELISA. A decrease in Th1 response with a diminishing in the relative expression of TBET and miR-29a at 2 and 6 months after the anti-Tb therapy (p < 0.01) were found. The miR-326 levels decreased after the intensive phase of the DOTS scheme. However, subdivision of the Tb patients according to gender, showed increased levels of miR-29a and miR-155 in females after the intensive phase of the therapeutic treatment when compared to time 0 and similar increased levels of miR-326 at time 6 versus time 0. In contrast, we observed a decrease in miR-326 levels in males at 6 months when compared to before therapy (time 0). In addition, high production of IL-17 in the culture supernatant was found at 2 and 6 months (p < 0.05) while in serum IL-17 was decreased. A positive correlation between IL-17 and RORC2 at time 6 was detected (p = 0.0202, r = 0.7880). In conclusion, these data suggest a reduction in Th1 and an induction of Th17 response after the anti-Tb therapy. © 2017 Elsevier Ltd
Tuberculosis (Tb) is an infectious disease in which the immune system plays an important role. MicroRNAs are involved in the development and maintenance of CD4 %2b T lymphocyte subpopulations. miR-326 regulates the differentiation to Th17 cells and miR-29 correlates with the Th1 response. The aim of this study was to determine the role of microRNAs, Transcription Factors, and cytokines in Th differentiation before and after the directly observed treatment short-course (DOTS). Peripheral blood mononuclear cells and serum from Tb patients were collected at times 0 (before therapy), 2 (after the intensive phase), and 6 months (after the holding phase). The cells were cultivated in presence or absence of ESAT-6 (10 μg/ml) and CFP-10 (10 μg/ml). Transcription Factor and microRNA expressions were analyzed by qPCR and cytokine production in both serum and culture supernatant using ELISA. A decrease in Th1 response with a diminishing in the relative expression of TBET and miR-29a at 2 and 6 months after the anti-Tb therapy (p < 0.01) were found. The miR-326 levels decreased after the intensive phase of the DOTS scheme. However, subdivision of the Tb patients according to gender, showed increased levels of miR-29a and miR-155 in females after the intensive phase of the therapeutic treatment when compared to time 0 and similar increased levels of miR-326 at time 6 versus time 0. In contrast, we observed a decrease in miR-326 levels in males at 6 months when compared to before therapy (time 0). In addition, high production of IL-17 in the culture supernatant was found at 2 and 6 months (p < 0.05) while in serum IL-17 was decreased. A positive correlation between IL-17 and RORC2 at time 6 was detected (p = 0.0202, r = 0.7880). In conclusion, these data suggest a reduction in Th1 and an induction of Th17 response after the anti-Tb therapy. © 2017 Elsevier Ltd
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Anti-Tb therapy; Cytokines; microRNAs; T cells; Transcription factors culture filtrate protein 10; cytokine; early secretory antigenic target 6; ethambutol plus isoniazid plus pyrazinamide plus rifampicin; gamma interferon; interleukin 10; interleukin 17; interleukin 4; microRNA; microRNA 146a; microRNA 155; microRNA 29a; microRNA 326; retinoid related orphan receptor gamma; retinoid related orphan receptor gamma 2; transcription factor; transcription factor GATA 3; transcription factor T bet; tumor necrosis factor; unclassified drug; cytokine; interleukin 17; microRNA; MIRN155 microRNA, human; MIRN29 microRNA, human; MIRN326 microRNA, human; retinoid related orphan receptor gamma; RORC protein, human; T box transcription factor; T-box transcription factor TBX21; transcription factor; tuberculostatic agent; adult; anti-infective therapy; Article; CD4 T lymphocyte; clinical article; cytokine production; enzyme linked immunosorbent assay; extrapulmonary tuberculosis; female; gene expression; human; human cell; lung tuberculosis; lymphocyte differentiation; male; middle aged; peripheral blood mononuclear cell; priority journal; prospective study; protein analysis; protein expression; short course therapy; Th1 cell; blood; cell culture; cell differentiation; directly observed therapy; drug effects; genetics; host pathogen interaction; immunology; metabolism; microbiology; Mycobacterium tuberculosis; Th1 cell; Th17 cell; time factor; treatment outcome; tuberculosis; Adult; Antitubercular Agents; Cell Differentiation; Cells, Cultured; Cytokines; Directly Observed Therapy; Female; Host-Pathogen Interactions; Humans; Interleukin-17; Male; MicroRNAs; Middle Aged; Mycobacterium tuberculosis; Nuclear Receptor Subfamily 1, Group F, Member 3; Prospective Studies; T-Box Domain Proteins; Th1 Cells; Th17 Cells; Time Factors; Transcription Factors; Treatment Outcome; Tuberculosis
Anti-Tb therapy; Cytokines; microRNAs; T cells; Transcription factors culture filtrate protein 10; cytokine; early secretory antigenic target 6; ethambutol plus isoniazid plus pyrazinamide plus rifampicin; gamma interferon; interleukin 10; interleukin 17; interleukin 4; microRNA; microRNA 146a; microRNA 155; microRNA 29a; microRNA 326; retinoid related orphan receptor gamma; retinoid related orphan receptor gamma 2; transcription factor; transcription factor GATA 3; transcription factor T bet; tumor necrosis factor; unclassified drug; cytokine; interleukin 17; microRNA; MIRN155 microRNA, human; MIRN29 microRNA, human; MIRN326 microRNA, human; retinoid related orphan receptor gamma; RORC protein, human; T box transcription factor; T-box transcription factor TBX21; transcription factor; tuberculostatic agent; adult; anti-infective therapy; Article; CD4+ T lymphocyte; clinical article; cytokine production; enzyme linked immunosorbent assay; extrapulmonary tuberculosis; female; gene expression; human; human cell; lung tuberculosis; lymphocyte differentiation; male; middle aged; peripheral blood mononuclear cell; priority journal; prospective study; protein analysis; protein expression; short course therapy; Th1 cell; blood; cell culture; cell differentiation; directly observed therapy; drug effects; genetics; host pathogen interaction; immunology; metabolism; microbiology; Mycobacterium tuberculosis; Th1 cell; Th17 cell; time factor; treatment outcome; tuberculosis; Adult; Antitubercular Agents; Cell Differentiation; Cells, Cultured; Cytokines; Directly Observed Therapy; Female; Host-Pathogen Interactions; Humans; Interleukin-17; Male; MicroRNAs; Middle Aged; Mycobacterium tuberculosis; Nuclear Receptor Subfamily 1, Group F, Member 3; Prospective Studies; T-Box Domain Proteins; Th1 Cells; Th17 Cells; Time Factors; Transcription Factors; Treatment Outcome; Tuberculosis
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