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

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• 作者：Phuong Mai Hoang (1)
  Seungchan Cho (1)
  Kee Eun Kim (1)
  Sung June Byun (2)
  Taek-Kyun Lee (3)
  Sukchan Lee (1)
  
  1. Department of Genetic Engineering ; Sungkyunkwan University ; 2066 Seobu-ro ; Jangan-gu ; Suwon ; 440-746 ; Korea
  2. Animal Biotechnology Division ; National Institute of Animal Science ; Rural Development Administration ; 143-13 Seosuwonro ; Gwonsun-gu ; Suwon ; 441-706 ; Korea
  3. South Sea Environment Research Department ; Korea Institute of Ocean Science and Technology ; Geoje ; 656-830 ; Korea
  
• 关键词：3D8 scFv ; Anti ; viral ; Lactobacillus paracasei ; Oral administration ; Murine norovirus
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2015
• 出版时间：March 2015
• 年：2015
• 卷：99
• 期：6
• 页码：2793-2803
• 全文大小：3,328 KB
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• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614

文摘

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.