The College of Veterinary Medicine, Henan Agricultural University, Nongye Road 63#, Zhengzhou, 450002, Henan Province, People's Republic of China.
Key Laboratory for Animal-Derived Food Safety of Henan Province, Zhengzhou, 450002, Henan, China.
Virol J. 2020 Oct 23;17(1):163. doi: 10.1186/s12985-020-01433-8.
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.
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, CD4IFN-γ, CD4IL-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.
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 CD4 and CD8 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 CD4IFN-γ T lymphocyte subset was the highest in the BPL group, and the positive rate of CD4IL-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.
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.
传染性胃肠炎病毒(TGEV)引起仔猪的肠内感染,其特征为呕吐、严重腹泻和脱水,而哺乳仔猪的死亡率通常高达 100%。接种疫苗是控制 TGEV 引起的疾病的有效措施。
本研究分别用甲醛(FA)、β-丙内酯(BPL)或双乙烯亚胺(BEI)对细胞培养的 TGEV HN-2012 株进行灭活,然后用弗氏佐剂制备灭活的 TGEV 疫苗,并在小鼠中评估其免疫效果。通过 ELISA 检测 TGEV 特异性 IgG 水平。通过流式细胞术检测 CD4、CD8、CD4IFN-γ、CD4IL-4 T 淋巴细胞的阳性率。还进行了淋巴细胞增殖试验以及大体病理和组织病理学检查,以评估三种不同的灭活试剂在制定 TGEV 疫苗中的作用。
结果表明,FA 组(n=17)的 TGEV 特异性 IgG 水平更早且更强,而 BEI 组产生的 IgG 水平则更持久。淋巴细胞增殖试验表明,BEI 组具有更强的诱导脾淋巴细胞增殖的能力。流式细胞术检测显示,BEI 组外周血淋巴细胞 CD4 和 CD8 T 淋巴细胞亚群的阳性率高于 FA 组和 BPL 组。BPL 组 CD4IFN-γ T 淋巴细胞亚群的阳性率最高,FA 组 CD4IL-4 T 淋巴细胞亚群的阳性率最高。接种疫苗的小鼠和对照组在宏观和组织病理学检查后均未见明显的病理变化。
这些结果表明,所有三组实验都可以诱导细胞和体液免疫,FA 组具有最佳的体液免疫效果,而 BEI 组则表现出优异的细胞免疫效果。