CAJ | 학술논문

应用荧光定量PCR技术,研究了嗜水气单胞菌疫苗浸泡免疫后,鳜(Siniperca chuatsi)鳃、皮肤、脾脏和头肾中IgM 基因表达量变化,同时应用ELISA检测皮肤黏液和血清中抗体滴度变化.结果显示:最早检测到Ig MmRNA转录水平上调的是皮肤和鳃(第4天),而脾脏和头肾在第7天才达到高峰.IgM基因在头肾和脾脏中表达量较高(高峰值分别达到16.3和23.8),皮肤和鳃中的表达量较小(高峰值分别为4.3和8.6).抗体效价方面,皮肤黏液中抗体滴度峰值出现时间较早(第7天),但抗体从开始形成到消失持续时间较短(28 d);血清中抗体滴度峰值出现时间较迟(第21天),但持续较长(42 d).结果表明:浸泡免疫能够引发鳜机体和局部均出现免疫应答.从抗体滴度和IgM基因表达量方面考虑,系统免疫组织均高于黏膜免疫组织,表明前者是合成IgM的主要场所.从时间上比较,对抗原刺激最早做出应答的是皮肤黏膜和鳃,其后系统免疫才表现应答作用,这表明了局部黏膜能够在抗原入侵的早期起到抵制作用.
응용형광정량PCR기술,연구료기수기단포균역묘침포면역후,궐(Siniperca chuatsi)새、피부、비장화두신중IgM 기인표체량변화,동시응용ELISA검측피부점액화혈청중항체적도변화.결과현시:최조검측도Ig MmRNA전록수평상조적시피부화새(제4천),이비장화두신재제7천재체도고봉.IgM기인재두신화비장중표체량교고(고봉치분별체도16.3화23.8),피부화새중적표체량교소(고봉치분별위4.3화8.6).항체효개방면,피부점액중항체적도봉치출현시간교조(제7천),단항체종개시형성도소실지속시간교단(28 d);혈청중항체적도봉치출현시간교지(제21천),단지속교장(42 d).결과표명:침포면역능구인발궐궤체화국부균출현면역응답.종항체적도화IgM기인표체량방면고필,계통면역조직균고우점막면역조직,표명전자시합성IgM적주요장소.종시간상비교,대항원자격최조주출응답적시피부점막화새,기후계통면역재표현응답작용,저표명료국부점막능구재항원입침적조기기도저제작용.
IgM gene expressions in Siniperca chuatsi immersed with inactivated Aeromonas hydrophilla were monitored by real-time quantitative PCR.And titers of antibody production in skin mucus and serum were studied by indirect ELISA.The results were as follows:raise of IgM mRNA transcription appeared to be primarily in the skin and gill (4 d) and secondarily in the head kidney and spleen (7 d).The gene expression levels of IgM were higher in the head kidney and spleen(peak value reached 16.3 and 23.8,respectively) than that in skin and gill (tiptop reached 4.3 and 8.6,respectively).Additionally,peak of antibody titer appeared on the 7th day and sustained 28 d in mucus and that of serum showed on the 21th day and lasted about 42 d.The results showed:immersion vaccination may induce systematic and local immune responses.Considering the expression amounts of IgM gene and titer,they were higher in lymphoid tissues than mucosa-associated tissues.It indicated that the lymphoid tissues were the major sites producing antibody.Judging from time,mucosa-associated tissues made immune responses in advance,followed by lymphoid tissues.The results support that mucosa-associated tissues play important roles in the early phase of antigen invasion.