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Murine Flt3 ligand-generated plasmacytoid and conventional dendritic cells display functional differentiation in activation, inflammation, and antigen presentation during BCG infection in vitro

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Abstract

Dendritic cells (DCs) are composed of distinct subsets. Their immunologic functions (especially in pathogenic infection, such as with mycobacteria) are poorly understood, largely because of their rarity and difficulty of preparation. We used the murine Fms-like tyrosine kinase 3 (Flt3) ligand to generate conventional DCs (FL-cDCs) and plasmacytoid DCs (FL-pDCs) and further evaluated their immunological responses to bacillus Calmette–Guérin (BCG) infection in vitro. BCG cells were observed inside both FL-cDCs and FL-pDCs by confocal microscopy, as confirmed by flow cytometric analysis showing a low infection rate of approximately 6 %, which was similar to in vivo results. The CD40, CD80, CD86, and MHC-II proteins were significantly upregulated in both FL-cDCs and -pDCs beginning at 4 h post-BCG exposure. FL-pDCs secreted TNF-α and IL-6 earlier and at significantly higher levels in the first 12 h following infection, but demonstrated delayed and weak activation and maturation compared to FL-cDCs. Although both subsets proved capable of presenting a mycobacterial antigen, FL-pDCs exhibited weaker activity in this respect than did FL-cDCs. In summary, the existence of FL-generated cDCs and pDCs imply functional differentiation in activation, inflammation, and antigen presentation, although both cells types participated extensively in the immune response to BCG infection.

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Acknowledgments

This work was supported by the National Basic Research and Development Program of China (2012CB518805), the National Natural Science Foundation of China (31372414), the Research and Development Program of Jiangsu (BE2015343), and the Priority Academic Development Program of Jiangsu Higher Education Institutions.

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Authors and Affiliations

  1. Jiangsu Key Laboratory of Zoonosis/Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, Jiangsu, China

    Chuang Meng, Xiaoyan Wang, Zhengzhong Xu, Maozhi Hu, Jiaying Liu, Zhiming Pan, Xiang Chen & Xinan Jiao

  2. Testing Center, Yangzhou University, Yangzhou, Jiangsu, China

    Maozhi Hu

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  1. Chuang Meng
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  2. Xiaoyan Wang
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  3. Zhengzhong Xu
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  4. Maozhi Hu
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  5. Jiaying Liu
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  6. Zhiming Pan
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  7. Xiang Chen
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  8. Xinan Jiao
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Correspondence to Xiang Chen or Xinan Jiao.

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Editor: Tetsuji Okamoto

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Meng, C., Wang, X., Xu, Z. et al. Murine Flt3 ligand-generated plasmacytoid and conventional dendritic cells display functional differentiation in activation, inflammation, and antigen presentation during BCG infection in vitro. In Vitro Cell.Dev.Biol.-Animal 53, 67–76 (2017). https://doi.org/10.1007/s11626-016-0076-3

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  • DOI: https://doi.org/10.1007/s11626-016-0076-3

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