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CD80是T细胞活化和增殖过程中不可缺少的膜抗原,为T细胞的增殖分化提供必要的共刺激信号,在特异性细胞免疫过程中具有重要作用。本研究从罗非鱼脾脏组织中克隆获得CD80基因的编码区序列(Gen Bank登录号:KU884324;命名为On-CD80),该基因全长810 bp,可编码269个氨基酸,理论分子量为29.83 k D,等电点为5.97。氨基酸序列分析显示On-CD80具有2个保守的免疫球蛋白样结构域(Ig-like domain),5'端有一个典型的信号肽序列,3'端具有1个跨膜结构域。系统进化树中罗非鱼与伯氏朴丽鱼CD80蛋白聚为一支,显示了较高的同源性。荧光定量PCR分析显示,On-CD80在健康尼罗罗非鱼组织中均有表达,且在脾脏组织中表达量最高。经灭活的无乳链球菌免疫后,On-CD80在肾脏、脾脏、胸腺、肠道、鳃、脑组织中的表达量均显著上调,且表现出较为明显的时间依赖性表达模式,肠道、鳃和脑分别在8 h、24 h出现表达高峰,并于48 h、96 h再次出现表达高峰。脾脏和肾脏、胸腺的On-CD80表达则分别是在48 h、72 h达到表达高峰。综上所述,On-CD80表达能被无乳链球菌所诱导,提示该分子可能参与罗非鱼的抗菌免疫应答,该结果为进一步研究On-CD80在罗非鱼感染无乳链球菌过程中的作用奠定了一定的基础。
Abstract：CD80 is an indispensable membrane antigen in the process of T cell activation and proliferation, which provides necessary co stimulatory signal for proliferation and differentiation of T cells and plays an important role in cell-mediated immunity. In this study, the coding sequence of CD80 gene was cloned from the spleen of tilapia(Gen Bank accession number: KU884324; designated as On-CD80). This gene was 810 bp in full length, encoding269 amino acids. The deduced amino acid sequence of On-CD80 with an estimated molecular weight of 29.83 k D and a theoretical p I of 5.97. Amino acid alignment indicated that it had two Ig-like domain conserved region. In 5'and 3' ends, there was a typical signal peptide sequence and a transmembrane domain respectively. Phylogenetic analysis showed that On-CD80 was clustered closely with Haplochromis burton, which indicated that the two species had high homology. Moreover, the m RNA expression levels in different tissues were analyzed by real time quantitative PCR. We found that On-CD80 could be detected in all the examined tissues in healthy Nile tilapia and the highest expression level was in the spleen. When immunized with inactivated Streptococcus agalactiae, the expression of On-CD80 significantly increased in kidney, spleen, thymus, intestine, gill and brain, and showed a time-dependent expression pattern of On-CD80. The highest level of expression in the intestine appeared at 8 h,and at 24 h in the gill and brain, and the second highest level reached at 48 h in intestine, and 96 h in the gill and brain. The highest On-CD80 expression level in the spleen and kidney occurred at 48 h, and at 72 h in the thymus.In summary, we forecasted that the expression of On-CD80 might be induced by Streptococcus agalactiae, which indicated that On-CD80 might be involved in the antibacterial immune response. In return, it might lay a foundation for the further research on what roles On-CD80 played in the immune response to Streptococcus agalactiae in the Nile tilapia.
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基本信息:
DOI：10.13417/j.gab.036.000190
中图分类号:S943
引用信息:
[1]汪志文,张海艳,黄瑜,等.尼罗罗非鱼CD80基因的分子克隆与mRNA表达分析[J].基因组学与应用生物学,2017,36(01):190-200.DOI:10.13417/j.gab.036.000190.
基金信息:
广东海洋大学创新强校项目(GDOU2013050108,GDOU2013050202);; 湛江市非资助性项目(2010C3107001)共同资助
2016-10-10
2016-10-10
2016-10-10