Assessments of different inactivating reagents in formulating transmissible gastroenteritis virus vaccine
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RESEARCH
Assessments of different inactivating reagents in formulating transmissible gastroenteritis virus vaccine Fujie Zhao1, Lintao Liu1, Menglong Xu1, Xiangli Shu1, Lanlan Zheng1* and Zhanyong Wei1,2*
Abstract Background: Transmissible gastroenteritis virus (TGEV) causes enteric infection in piglets, characterized by vomiting, severe diarrhea and dehydration, and the mortality in suckling piglets is often high up to 100%. Vaccination is an effective measure to control the disease caused by TGEV. Methods: In this study, cell-cultured TGEV HN-2012 strain was inactivated by formaldehyde (FA), β-propiolactone (BPL) or binaryethylenimine (BEI), respectively. Then the inactivated TGEV vaccine was prepared with freund’s adjuvant, and the immunization effects were evaluated in mice. The TGEV-specific IgG level was detected by ELISA. The positive rates of CD4+, CD8+, CD4+IFN-γ+, CD4+IL-4+ T lymphocytes were detected by flow cytometry assay. Lymphocyte proliferation assay and gross pathology and histopathology examination were also performed to assess the three different inactivating reagents in formulating TGEV vaccine. Results: The results showed that the TGEV-specific IgG level in FA group (n = 17) was earlier and stronger, while the BEI group produced much longer-term IgG level. The lymphocyte proliferation test demonstrated that the BEI group had a stronger ability to induce spleen lymphocyte proliferation. The positive rates of C D4+ and C D8+ T lymphocyte subsets of peripheral blood lymphocyte in BEI group was higher than that in FA group and BPL groups by flow cytometry assay. The positive rate of CD4+IFN-γ+ T lymphocyte subset was the highest in the BPL group, and the positive rate of CD4+IL-4+ T lymphocyte subset was the highest in the FA group. There were no obvious pathological changes in the vaccinated mice and the control group after the macroscopic and histopathological examination. Conclusions: These results indicated that all the three experimental groups could induce cellular and humoral immunity, and the FA group had the best humoral immunity effect, while the BEI group showed its excellent cellular immunity effect. Keywords: Transmissible gastroenteritis virus (TGEV), Formaldehyde, β-propiolactone, Binary-ethylenimine, Inactivated vaccine
*Correspondence: [email protected]; [email protected] 1 The College of Veterinary Medicine, Henan Agricultural University, Nongye Road 63#, Zhengzhou 450002, Henan Province, People’s Republic of China Full list of author information is available at the end of the article
Introduction Transmissible gastroenteritis virus (TGEV) is an enveloped, positive, single-stranded RNA virus, which belongs to the Alphacoronavirus genus, Coronaviridae family. TGEV causes acute enteric disease in pigs, characterized by vomiting, severe diarrhea and dehydration. The mortality of TGEV often reaches 100% in suckling piglets less than two weeks of age, and causes huge economic losses in pig industry around the world [1, 2]. Until now, there
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