非致病性大肠杆菌鞭毛蛋白对O型口蹄疫病毒的佐剂效果

doi:10.3864/j.issn.0578-1752.2017.09.017

Abstract

Abstract: 【Objective】Foot and mouth disease (FMD) is highly infections and spread wide. It has brought a great harm to the world economic and social development. The current vaccination is still the main and effective means of prevention and control of this disease. To enhance FMD Virus (FMDV) antigenicity, the Flagella (F0) was modified from the non-pathogenic Escherichia coli to construct recombinant F0 protein and evaluate the single F0 protein and fusion protein F0-LTMT adjuvant effectiveness on FMDV based on the research about flagella protein adjuvants from home and abroad.【Method】Four recombinant proteins of F0, F0-LTMT, F0NC, and F0NC-LTMT were expressed, and their biological activities were detected by GM1-ELISA. The FMDV immunogen was prepared by mixing inactivated FMDV with a single recombinant F0 at 5 μg per mouse, and additional ISA 206 adjuvant was set according to the proportion to evaluate the immune effect in mice. The Balb/c mice was subcutaneously inoculated with emulsified immunogen twice at intervals of 2 weeks. Sera and feces were collected, respectively, on the day 0, 14, 21, 28, 35 and 45, and distal intestine was also collected after immunization on day 45. Titers of IgG in serum were detected by blocking ELISA kit, specific sIgA in feces and intestinal liquid by indirect ELISA to evaluate the titers.【Result】SDS-PAGE and Western blot analyses showed that four different recombinant flagellin were successfully obtained. Their biological activities were verified by GM l-ELISA. In mice no significant levels of antibodies were detected in the single FMDV immunized group compared with higher IgG titer sIgA titers in flagellin adjuvant group. Moreover, the synergistic effect of LTMT and flagellin facilitated to produce more IgG and sIgA in mice. When flagellin combined with the ISA 206 adjuvant, the titer of serum IgG increased significantly and gave extended duration. The mixed immunogen of FMDV, ISA 206 and H0NC-LTMT displayed the best antibody titers.【Conclusion】F0-LTMT provided dual adjuvant effectiveness to FMDV, induced systemic specific IgG and sIgA in mice. It is a potential mucosal adjuvant for FMDV.

Key words: FMDV, Escherichia coli, flagellin, mucosal adjuvant

SUN XiaoHan, ZHANG BiCheng, ZHANG Qiang, HE KongWang, ZHANG XueHan. Adjuvant Effects of Flagellin from Non-Pathogenic E.coli on FMDV[J].Scientia Agricultura Sinica, 2017, 50(9): 1714-1722.

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