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The recombinant fusion protein of cholera toxin B and neutrophil-activating protein expressed on Bacillus subtilis spore surface suppresses allergic inflammation in mice

  • Applied microbial and cell physiology
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Abstract

The neutrophil-activating protein of Helicobacter pylori (HP-NAP) has been identified as a modulator with anti-Th2 inflammation activity, and cholera toxin B (CTB) is a mucosal adjuvant that can also induce antigen tolerance. In this study, we constructed a CTB-NAP fusion protein on the surface of Bacillus subtilis spore and evaluate the efficiency of oral administration of the recombinant CTB-NAP spores in preventing asthma in mice. Oral administration of recombinant CTB or CTB-NAP spores significantly decreased serum ovalbumin (OVA)-specific IgE (p < 0.001) and increased fecal IgA (p < 0.01) compared to the treatment with non-recombinant spores. Oral administration of recombinant CTB or CTB-NAP spores induced IL-10 and IFN-γ expression and reduced IL-4 levels in bronchoalveolar lavage fluid (BALF). Moreover, CTB and CTB-NAP spores reduced the eosinophils in BALF and inflammatory cell infiltration in the lungs. Furthermore, CD4+CD25+Foxp3+ Tregs in splenocytes were significantly increased in mice treated with recombinant CTB or CTB-NAP spores. The number of CD4+CD25+Foxp3+ Tregs caused by CTB-NAP was higher than that by CTB alone. Our study indicated that B. subtilis spores with surface expression of subunit CTB or CTB-NAP could inhibit OVA-induced allergic inflammation in mice. The attenuated inflammation was attributed to the induction of CD4+CD25+Foxp3+ Tregs and IgA. Moreover, the fusion protein CTB-NAP demonstrated a better efficiency than CTB alone in inhibiting the inflammation.

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Acknowledgements

This study was founded by the Guangdong Science and Technology Foundation (no. 2014A020212013 and no. 2016A020215013), Natural Science Foundation of Guangdong (no. 8451012001001570 and no. 915101200F009), National Natural Science Foundation of China (no. 30801054), and Science and Technology Foundation of Guangzhou (no. 201707010010).

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  1. Hui Dong and Yanmei Huang contributed equally to the article.

Authors and Affiliations

  1. Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou, 510120, China

    Hui Dong, Yanmei Huang, Shuwen Yao, Bingshao Liang, Yan Long, Yongqiang Xie, Jialiang Mai, Sitang Gong & Zhenwen Zhou

  2. The First Women and Children’s Hospital of Huizhou, Huizhou, 516000, China

    Hui Dong

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  1. Hui Dong
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Dong, H., Huang, Y., Yao, S. et al. The recombinant fusion protein of cholera toxin B and neutrophil-activating protein expressed on Bacillus subtilis spore surface suppresses allergic inflammation in mice. Appl Microbiol Biotechnol 101, 5819–5829 (2017). https://doi.org/10.1007/s00253-017-8370-x

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