摘要
将大肠杆菌重组表达的鳜(Siniperca chuatsi)传染性脾肾坏死病毒(ISKNV)主衣壳蛋白(MCP),以不同剂量(20μg/尾、50μg/尾、100μg/尾)腹腔注射免疫幼鳜(2月龄),测定各免疫组的血清抗体效价变化规律、头肾淋巴细胞增殖刺激指数(SI)、肝脏Mx蛋白表达及相对免疫保护率(RPS)。结果表明,各免疫组抗体效价在第14天时达到峰值,其中50μg/尾免疫组的抗体效价最高,随后各组的效价均下降,至第35天时与对照组无差异;头肾淋巴细胞经LPS、ConA刺激后,50μg/尾免疫组及100μg/尾免疫组的刺激指数均升高,其中50μg/尾免疫组的刺激指数最高,20μg/尾免疫组与对照组无差异;在免疫后48h各剂量免疫组Mx蛋白均有低量表达,对照组无表达,各免疫组表达量无显著差异;免疫后第36天攻毒,50μg/尾免疫组的相对保护率最高,为64.3%。研究结果表明,重组MCP蛋白有较好的免疫原性,可以激发鳜特异性免疫及非特异性免疫应答,当免疫剂量为50μg/尾时,免疫保护效果最好。
Infectious spleen and kidney necrosis virus（ ISKNV） from mandarin fish, Siniperca chuatsi, is the causative agent of a disease causing high mandarin fish mortalities and leading to great economic losses in China. In this study, to develop a vaccine to protect mandarin fish against infection with ISKNV, the immunogenicity of the recombinant major capsid protein（MCP） was tested with several experiments. The juvenile Mandarin fish were vaccinated by intraperitoneal injection with MCP expressed in E. coli, and the dose was 20 μg/ind,50 μg/ind and 100 μg/ind respectively. Several immunological indexes were measured which include the kinetics of serum antibody, the stimulation index（SI） of the head kidney lymphocyte proliferation, the expression of Mx（myxovirus resistance） protein in liver and the relative percentage survival（RPS）.The results showed that the antibody titer of the 50μg/ind group was higher than the others, and on the 14th day that of each vaccination group reached the peak, then dropped down, on the 35th day that of each vaccination group showed no difference from the control group. After head kidney lymphocyte of each group stimulated with LPS and ConA, respectively, the SI of 50 μg/ind group was the highest and that of 20 μg/ind group showed no difference from the control group. The Mx protein expressed low-levelly at 48 h post-vaccination, and its expression showed no significant difference in three vaccination groups. After virulent challenge infection, the RPS of 50 μg/ind group was the highest,accounting for 64.3%. In conclusion, the best specific and nonspecific immunity of Siniperca chuatsi were induced by renatured MCP with dose of 50 μg/ind.
出处
《中国水产科学》
CAS
CSCD
北大核心
2009年第3期388-393,共6页
Journal of Fishery Sciences of China
基金
国家科技支撑计划资助项目(2006BAD03B05)
广东省自然科学基金资助项目(7004728).
关键词
鳜
传染性脾肾坏死病毒
主衣壳蛋白
免疫保护
Siniperca chuatsi
infectious spleen and kidney necrosis viru（sISKNV）
major capsid protein（MCP）
immune protection
