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目的:通过研究EIAV弱毒疫苗株和强毒株在诱导多种特异性抗体方面的差异,初步探讨特异性体液免疫应答在EIAV弱毒疫苗株保护性免疫中的作用。方法:试验马匹根据接种毒株分为疫苗组和强毒隐性感染组,应用ELISA方法对血清中病毒囊膜蛋白Env和衣壳蛋白P26结合抗体的滴度、亲和力及Env结合抗体的构象依赖性进行动态实相检测,同时采用细胞蚀斑染色和逆转录酶活性检测方法对中和抗体水平进行同期检测。结果:Env和P26结合抗体的滴度、亲和力、构象依赖性及中和抗体的中和活性均存在一个缓慢上升的过程。对于P26抗体滴度及Env抗体亲和力,疫苗组和强毒隐性感染组仅在病毒接种2个月内,表现为统计学差异(P<0.05)。疫苗组从接种后14d开始,Env抗体的构象依赖性高于强毒隐性感染组(P<0.05),并持续至最后一个监测点。最明显的是,疫苗组诱导产生具有中和强毒特性毒株(EIAVDLV34)和弱毒特性毒株(EIAVFDDV)活性的中和抗体均高于强毒隐性感染组(P<0.05)。结论:弱毒疫苗株和强毒株诱导机体产生结合抗体和中和抗体的时间、水平和性质均存在明显差异,其中中和抗体和构象依赖性抗体的差异,可能是EIAV弱毒疫苗诱导保护性免疫的主要因素之一。
OBJECTIVE: To investigate the role of specific humoral immune responses in protective immunity against EIAV attenuated vaccine strains by studying the differences in the induction of multiple specific antibodies between attenuated and attenuated EIAV strains. Methods: The test animals were divided into vaccine group and virulent latent infection group according to the inoculated strains. The titer and affinity of the virus envelope protein Env and the capsid protein P26-binding antibody in serum and the conformational dependence of Env-binding antibody Sexual real-time detection, at the same time using cell plaque staining and reverse transcriptase activity detection method of neutralizing antibody levels for simultaneous detection. RESULTS: The titer, affinity, conformational dependence, and the neutralizing activity of neutralizing antibodies of Env and P26-bound antibodies all had a slowly increasing process. For the P26 antibody titer and Env antibody affinity, the vaccine group and virulent latent infection group showed statistical difference (P <0.05) only within 2 months after virus inoculation. From the 14th day after inoculation, Env antibody conformational dependence was higher in the vaccine group than in the virulent latent infection group (P <0.05), and continued until the last monitoring point. Most notably, the neutralizing antibodies induced by the vaccine group that produced both EIAVDLV34 and EIAVFDDV were significantly higher than those of the virulent HTV group (P <0.05). Conclusions: The time, level and nature of attenuated and neutralizing antibodies induced by attenuated vaccine strains and virulent strains are obviously different. The differences of neutralizing antibodies and conformation-dependent antibodies may be caused by the protective immunity induced by attenuated EIAV vaccine One of the main factors.