Construction and characterization of Lactobacillus pentosus expressing the D antigenic site of the spike protein of Transmissible gastroenteritis virus

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Abstract:

This study explored the feasibility of Lactobacillus pentosus as a live vehicle to deliver and express antigen. First of all, L. pentosus transformed by electroporation with the plasmids pg611-6D (anchored) and pg612-6D (secretory) based on the xylose operon generated the recombinant strains rLppg611-6D and rLppg612-6D, respectively, expressing the D antigenic site of the spike (S) protein of Transmissible gastroenteritis virus (TGEV), for intragastric administration in mice. Secondly, we collected serum, fecal, nasal, ophthalmic, and vaginal samples from pre-immune mice and after the first immunization (on days 7, 14, 21, 28, 35, and 42) that were used to analyze the levels of immunoglobulins G and A against TGEV by using ELISA. In addition, a plaque reduction assay was performed using sera from groups pg611, pg612-6D, pg11-6D, and phosphate-buffered saline (blank control) to analyze TGEV-neutralizing antibody activity in vitro. A statistically significant difference in serum tests between groups demonstrated that rLppg612-6D induced better immunogenicity than rLppg611-6D, making rLppg612-6D the better candidate for oral vaccine. Taken together, L. pentosus possessed the potential to become a novel vector for mucosal vaccine in the future.

Keywords: Lactobacillus pentosus; S protein D site; Transmissible gastroenteritis virus; site D de la protéine S; virus de la gastroentérite transmissible

Document Type: Research Article

DOI: http://dx.doi.org/10.1139/w11-027

Affiliations: Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, Harbin 150030, the People’s Republic of China.

Publication date: May 4, 2011

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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