Induction of transcription factors, miRNAs and cytokines involved in T lymphocyte differentiation in BCG-vaccinated subjects
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The BCG vaccine induces a Th1 phenotype, which is essential for protection against Mycobacterium tuberculosis. However, the effects of BCG vaccination over time on the T helper subpopulation and the microRNAs involved in adulthood have not been studied. In the present study, we explored the involvement of microRNAs, transcription factors and multifunctional cytokines in BCG vaccination by examining their levels both before and after vaccination of healthy adults. Peripheral blood mononuclear cells were obtained at 0, 2 and 6 months after vaccination. Cells were cultured in the presence or absence of ESAT-6-6 and CFP-10-10 or M. tuberculosis filtrate. The expression levels of miRNAs and transcription factors were evaluated using qRT-PCR. Cytokine production in supernatants and serum samples was evaluated using ELISA. Multifunctional CD4 T cells were analyzed using multiparametric flow cytometry. We observed a decrease in the expression levels of T-BET, GATA3 and FOXP3 at 2 months and miR-146a, miR-326 and miR-155 at 6 months after receiving the vaccine. In the supernatant, the production of IL-17 was increased after 6 months, with both stimuli. In contrast, IL-10, TNF-α and IFN-γ increased at 2 months. In the serum, high levels of IL-10 were found after 2 months compared to time 0 and 6 months. The production of multifunctional cells that expressed the cytokine profiles CD4 TNF-α IFN-γ-IL-10-, CD4 TNF-α IL-1IFN-γ-, CD4 IL-10 IFN-γ-TNF-α- and CD4 IL-17 IFN-γ- predominantly increased after 2 months with and without the stimulus. Correlation analysis revealed a negative association between FOXP3 and miR-155 (r = −0.5120, p = 0.0176) and between IL-17 and miR-326 (r = −0.5832, p = 0.0364). This study is the first to demonstrate roles for microRNAs, transcription factors and cytokines in the T helper differentiation lineage and to describe the possible mechanism by which their expression is modulated by the presence of the BCG vaccine in adulthood. In conclusion, our results suggest that the BCG vaccine induces a modulation in transcription factors and miRNAs with high production of multifunctional cells CD4 TNF-α IL-10 IFN-γ-. © 2016 Elsevier Ltd
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The BCG vaccine induces a Th1 phenotype, which is essential for protection against Mycobacterium tuberculosis. However, the effects of BCG vaccination over time on the T helper subpopulation and the microRNAs involved in adulthood have not been studied. In the present study, we explored the involvement of microRNAs, transcription factors and multifunctional cytokines in BCG vaccination by examining their levels both before and after vaccination of healthy adults. Peripheral blood mononuclear cells were obtained at 0, 2 and 6 months after vaccination. Cells were cultured in the presence or absence of ESAT-6-6 and CFP-10-10 or M. tuberculosis filtrate. The expression levels of miRNAs and transcription factors were evaluated using qRT-PCR. Cytokine production in supernatants and serum samples was evaluated using ELISA. Multifunctional CD4%2b T cells were analyzed using multiparametric flow cytometry. We observed a decrease in the expression levels of T-BET, GATA3 and FOXP3 at 2 months and miR-146a, miR-326 and miR-155 at 6 months after receiving the vaccine. In the supernatant, the production of IL-17 was increased after 6 months, with both stimuli. In contrast, IL-10, TNF-α and IFN-γ increased at 2 months. In the serum, high levels of IL-10 were found after 2 months compared to time 0 and 6 months. The production of multifunctional cells that expressed the cytokine profiles CD4%2bTNF-α%2bIFN-γ-IL-10-, CD4%2bTNF-α%2bIL-1IFN-γ-, CD4%2bIL-10%2bIFN-γ-TNF-α- and CD4%2bIL-17%2bIFN-γ- predominantly increased after 2 months with and without the stimulus. Correlation analysis revealed a negative association between FOXP3 and miR-155 (r = −0.5120, p = 0.0176) and between IL-17 and miR-326 (r = −0.5832, p = 0.0364). This study is the first to demonstrate roles for microRNAs, transcription factors and cytokines in the T helper differentiation lineage and to describe the possible mechanism by which their expression is modulated by the presence of the BCG vaccine in adulthood. In conclusion, our results suggest that the BCG vaccine induces a modulation in transcription factors and miRNAs with high production of multifunctional cells CD4%2bTNF-α%2bIL-10%2bIFN-γ-. © 2016 Elsevier Ltd
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BCG vaccination; MicroRNAs; Multifunctional cells; Transcription factors; Treg cells APC protein; BCG vaccine; culture filtrate protein 10; cytokine; early secretory antigenic target 6; gamma interferon; interleukin 1; interleukin 10; interleukin 17; microRNA; microRNA 146a; microRNA 155; microRNA 29; microRNA 29a; microRNA 326; retinoid related orphan receptor gamma; transcription factor; transcription factor FOXP3; transcription factor GATA 3; transcription factor T bet; tumor necrosis factor; unclassified drug; BCG vaccine; cytokine; forkhead transcription factor; FOXP3 protein, human; gamma interferon; IL10 protein, human; interleukin 10; microRNA; MIRN146 microRNA, human; MIRN155 microRNA, human; MIRN326 microRNA, human; T box transcription factor; T-box transcription factor TBX21; transcription factor; adult; Article; BCG vaccination; CD4 T lymphocyte; cell culture; cell lineage; comparative study; controlled study; cytokine production; enzyme linked immunosorbent assay; female; flow cytometry; gene expression; human; human cell; immunomodulation; lymphocyte differentiation; male; Mycobacterium tuberculosis; normal human; peripheral blood mononuclear cell; priority journal; protein blood level; protein expression; reverse transcription polymerase chain reaction; T lymphocyte; tuberculin test; tuberculosis; adolescent; biosynthesis; cell differentiation; immunology; polymerase chain reaction; young adult; Adolescent; BCG Vaccine; CD4-Positive T-Lymphocytes; Cell Differentiation; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Forkhead Transcription Factors; Humans; Interferon-gamma; Interleukin-10; Male; MicroRNAs; Polymerase Chain Reaction; T-Box Domain Proteins; Transcription Factors; Young Adult
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BCG vaccination; MicroRNAs; Multifunctional cells; Transcription factors; Treg cells APC protein; BCG vaccine; culture filtrate protein 10; cytokine; early secretory antigenic target 6; gamma interferon; interleukin 1; interleukin 10; interleukin 17; microRNA; microRNA 146a; microRNA 155; microRNA 29; microRNA 29a; microRNA 326; retinoid related orphan receptor gamma; transcription factor; transcription factor FOXP3; transcription factor GATA 3; transcription factor T bet; tumor necrosis factor; unclassified drug; BCG vaccine; cytokine; forkhead transcription factor; FOXP3 protein, human; gamma interferon; IL10 protein, human; interleukin 10; microRNA; MIRN146 microRNA, human; MIRN155 microRNA, human; MIRN326 microRNA, human; T box transcription factor; T-box transcription factor TBX21; transcription factor; adult; Article; BCG vaccination; CD4+ T lymphocyte; cell culture; cell lineage; comparative study; controlled study; cytokine production; enzyme linked immunosorbent assay; female; flow cytometry; gene expression; human; human cell; immunomodulation; lymphocyte differentiation; male; Mycobacterium tuberculosis; normal human; peripheral blood mononuclear cell; priority journal; protein blood level; protein expression; reverse transcription polymerase chain reaction; T lymphocyte; tuberculin test; tuberculosis; adolescent; biosynthesis; cell differentiation; immunology; polymerase chain reaction; young adult; Adolescent; BCG Vaccine; CD4-Positive T-Lymphocytes; Cell Differentiation; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Forkhead Transcription Factors; Humans; Interferon-gamma; Interleukin-10; Male; MicroRNAs; Polymerase Chain Reaction; T-Box Domain Proteins; Transcription Factors; Young Adult
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