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Development of Lactobacillus paracasei harboring nucleic acid-hydrolyzing 3D8 scFv as a preventive probiotic against murine norovirus infection

  • Applied microbial and cell physiology
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Abstract

The protein 3D8 single-chain variable fragment (3D8 scFv) has potential anti-viral activity due to its ability to penetrate into cells and hydrolyze nucleic acids. Probiotic Lactobacillus paracasei engineered to secrete 3D8 scFv for oral administration was used to test the anti-viral effects of 3D8 scFv against gastrointestinal virus infections. We found that injection of 3D8 scFv into the intestinal lumen resulted in the penetration of 3D8 scFv into the intestinal villi and lamina propria. 3D8 scFv secreted from engineered L. paracasei retained its cell-penetrating and nucleic acid-hydrolyzing activities, which were previously shown with 3D8 scFv expressed in Escherichia coli. Pretreatment of RAW264.7 cells with 3D8 scFv purified from L. paracasei prevented apoptosis induction by murine norovirus infection and decreased messenger RNA (mRNA) expression of the viral capsid protein VP1. In a mouse model, oral administration of the engineered L. paracasei prior to murine norovirus infection reduced the expression level of mRNA encoding viral polymerase. Taken together, these results suggest that L. paracasei secreting 3D8 scFv provides a basis for the development of ingestible anti-viral probiotics active against gastrointestinal viral infection.

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Acknowledgments

This study was supported by a grant from the Agenda (No. PJ0102012014) from the Rural Development Administration (RDA) of Korea and a grant from the Korea Institute of Ocean Science and Technology project (No. PE99154). The authors wish to thank Dr. Jos Seegers (Falcobio, Netherlands) for providing the Lactobacillus casei strain and expression vector, as well as for providing valuable advice. We also wish to thank Dr. Jai Myung Yang for providing RAW264.7 cells and murine norovirus.

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Authors and Affiliations

  1. Department of Genetic Engineering, Sungkyunkwan University, 2066 Seobu-ro, Jangan-gu, Suwon, 440-746, Korea

    Phuong Mai Hoang, Seungchan Cho, Kee Eun Kim & Sukchan Lee

  2. Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, 143-13 Seosuwonro, Gwonsun-gu, Suwon, 441-706, Korea

    Sung June Byun

  3. South Sea Environment Research Department, Korea Institute of Ocean Science and Technology, Geoje, 656-830, Korea

    Taek-Kyun Lee

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  1. Phuong Mai Hoang
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  2. Seungchan Cho
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Correspondence to Sukchan Lee.

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Hoang, P.M., Cho, S., Kim, K.E. et al. Development of Lactobacillus paracasei harboring nucleic acid-hydrolyzing 3D8 scFv as a preventive probiotic against murine norovirus infection. Appl Microbiol Biotechnol 99, 2793–2803 (2015). https://doi.org/10.1007/s00253-014-6257-7

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  • DOI: https://doi.org/10.1007/s00253-014-6257-7

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