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Adjuvant Effects of Deleting Hypervariable Domains of FliC of Escherichia coli Nissle 1917

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Abstract

Bacterial flagellin (FliC) can be used as a TLR5 ligand-like adjuvant. However, the sequences of hypervariable regions(HVR) of FliC from different bacteria vary, and their effects on adjuvants remain unclear. In this study, FliCΔ274–406 (deleting of D3 domain) and FliCΔ174–506 (deleting of D2-D3 domain) from Escherichia coli Nissle 1917 FliC (FliCEcN) were constructed and expressed in host bacteria, BL21. Purification was conducted using affinity chromatography on a Ni-NTA column, validation was done using SDS-PAGE and western blotting, the antigenicity and immunogenicity were detected using ELISA, and adjuvant effects were evaluated in Caco-2 cells and mice. The results showed that FliCEcN was mainly expressed in the bacterial supernatant, and the two truncated flagellins were expressed as inclusion bodies. Compared with FliCEcN, both FliCΔ274–406 and FliCΔ174–506 had considerable decreased antigenicity and immunogenicity. In Caco-2 cells, FliCΔ174–506 had a higher ability to promote the secretion of IL-8 than FliCEcN and FliCΔ274–406. In mice, FliCΔ174–506 showed a comparable adjuvant level to FliCEcN, while FliCΔ274–406 was less effective. Our data shows that adjuvant effects of FliCEcN with deletion of different regions of its HVR are inconsistent, and deleting its entire D2–D3 domain is better.

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DATA AVAILABILITY

The datasets generated in the present study are available from the authors upon request.

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ACKNOWLEDGMENTS

The authors would like to be very thankful to the College for Veterinary Medicine, Guizhou University, China for supporting and helping to complete this research work timely. At the same time, we thank all the contributors who provided financial support for this project.

Funding

The project was supported by the National Natural Science Foundation of China (Grant no. 32102703), Guangdong Basic and Applied Basic Research Foundation (2019A1515111186), Guizhou Provincial Science and Technology Projects (Qian Ke He Basic_ZK[2023] General 105), the Plan of Science and Technology of Guizhou Province (Qian Ke He LH Zi [2017] no. 7268), the project of Introducing Talents in Guizhou University (GDRJHZ[2016]77), the Support Plan Project of Science and Technology of Guizhou Province in 2021 (Qian Ke He Support [2021 ] General 164).

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Authors and Affiliations

  1. Guizhou University, College of Veterinary Medicine, 550025, Guizhou, China

    Shuang Li, Yongjun Deng, Ming Wen, Ying Yang & Guilan Wen

  2. School of Life Sciences, Zhaoqing University, Zhaoqing, 526000, China, China

    Bingming Ou

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  1. Shuang Li
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  2. Yongjun Deng
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  3. Bingming Ou
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  4. Ming Wen
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  5. Ying Yang
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  6. Guilan Wen
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Contributions

Conceptualization: Ying Yang, Ming Wen, Guilan Wen; Data curation: Bingming Ou; Formal analysis: Yongjun Deng; Funding acquisition: Ying Yang, Ming Wen; Methodology: Yongjun Deng; Project administration: Ying Yang; Supervision: Ying Yang, Ming Wen, Guilan Wen; Validation: Shuang Li; Writing—original draft: Shuang Li; Writing—review and editing: Shuang Li, Ying Yang.

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Correspondence to Ying Yang.

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Li, S., Deng, Y., Ou, B. et al. Adjuvant Effects of Deleting Hypervariable Domains of FliC of Escherichia coli Nissle 1917. Mol. Genet. Microbiol. Virol. 38, 207–214 (2023). https://doi.org/10.3103/S0891416823030047

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