ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2017, Vol. 48 ›› Issue (5): 914-921.doi: 10.11843/j.issn.0366-6964.2017.05.016

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Cyclic GMP-AMP Synthase-mediated Maturation of Murine Bone Marrow Derived Dendritic Cells after Infection with Mycobacterium bovis

LI Qiang1, LIU Chun-fa2, HE Xiao-li1, LI Fan-fei1, WEI Fan-hua1, ZHAO De-ming2, ZHOU Xiang-mei2*, XU Li-hua1*   

  1. 1. College of Agriculture, Ningxia University, Yinchuan 750021, China;
    2. National Animal Transmissible Spongiform Encephalopathy Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
  • Received:2016-09-21 Online:2017-05-23 Published:2017-05-23

Abstract:

The study was performed to investigate the regulation of cyclic GMP-AMP synthase (cGAS) on Mycobacterium bovis infected murine bone marrow derived dendritic cell (BMDC). The murine bone marrow cells were cultured and induced into DCs in vitro. Then BMDCs were infected by Mycobacterium bovis to establish infection models. Three groups were designed for the in vitro study. They were control group (BMDC control), infection group (M. bovis infected BMDC) and silence group. In silence group, BMDC cGAS gene was knocked down via siRNA and then cells were infected similar to infection group. Flow cytometry was used to analyze the expression of surface markers. In cGAS pathway, STING, TBK1, p-TBK1 and IRF3 were determined by Western blot and Immunofluorescence, and the cytokine production in cell supernatant was assessed by ELISA. The data showed that M. bovis induced higher expression levels of the surface markers CD86, CD80, CD40, MHC-Ⅱ and cytokine production IFN-β, TNF-α, IL-12p70, IL-6 in BMDC. Meanwhile, the cGAS pathway was activated and induced higher expression of related protein, however corresponding data reduced significantly in silence group. Our research indicated that cGAS pathway can be activated by Mycobacterium bovis when BMDC was infected, and cGAS contribute to maturation and activation of BMDC.

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