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尼罗罗非鱼Igλ单克隆抗体的制备及尼罗罗非鱼Igλ B细胞亚群研究

Preparation of the monoclonal antibody against Nile tilapia Igλ and study on the Igλ B cell subset in Nile tilapia.

作者信息

Song Yan-Ling, Yao Yuan-Yuan, Liu Xun, Tian Tian-Tian, Ye Jian-Min, Zhang Yong-An, Zhang Xu-Jie

机构信息

State Key Laboratory of Agricultural Microbiology, Hubei Hongshan Laboratory, Engineering Research Center of Green Development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt, Ministry of Education, Huazhong Agricultural University, Wuhan, China.

Key Laboratory for Feed Biotechnology of the Ministry of Agriculture and Rural Affairs, Institute of Feed Research, Chinese Academy of Agricultural Sciences, Beijing, China.

出版信息

Fish Shellfish Immunol. 2023 May;136:108705. doi: 10.1016/j.fsi.2023.108705. Epub 2023 Mar 21.

Abstract

Immunoglobulins (Igs) are important effector molecules that mediate humoral immunity. A typical Ig consists of two heavy and two light chains. In teleosts, three Ig heavy chain isotypes (Igμ, Igδ and Igτ) and three Ig light chain isotypes (Igκ, Igλ and Igσ) have been identified. Compared to the heavy chains, teleost Ig light chains have been poorly studied due to the lack of antibodies. In this study, a mouse anti-Nile tilapia Igλ monoclonal antibody (mAb) was prepared, which could specifically recognize Igλ in serum and Igλ B cells in tissues. Further, the composition of IgM and Igλ B cell subsets was analyzed using this antibody and a mouse anti-tilapia IgM heavy chain mAb. The ratio of IgMIgλ B cells to total IgM B cells in head kidney and peripheral blood was about 30%, while that in spleen was about 50%; the ratio of IgMIgλ B cells to total Igλ B cells in head kidney and peripheral blood was about 45%, while that in spleen was about 25%. The IgMIgλ B cells was speculated to be IgT B cells. Finally, we detected an increase in the level of specific antibodies against the surface antigen-Sip of Streptococcus agalactiae in serum after S. agalactiae infection, indicating that mouse anti-tilapia Igλ mAb can be used to detect the antibody level after immunization of Nile tilapia, which lays a foundation for the evaluation of immunization effect of tilapia vaccine.

摘要

免疫球蛋白(Igs)是介导体液免疫的重要效应分子。典型的免疫球蛋白由两条重链和两条轻链组成。在硬骨鱼中,已鉴定出三种免疫球蛋白重链同种型(Igμ、Igδ和Igτ)和三种免疫球蛋白轻链同种型(Igκ、Igλ和Igσ)。与重链相比,由于缺乏抗体,硬骨鱼免疫球蛋白轻链的研究较少。在本研究中,制备了一种小鼠抗尼罗罗非鱼Igλ单克隆抗体(mAb),该抗体可特异性识别血清中的Igλ和组织中的Igλ B细胞。此外,使用该抗体和小鼠抗罗非鱼IgM重链单克隆抗体分析了IgM和Igλ B细胞亚群的组成。头肾和外周血中IgMλ B细胞与总IgM B细胞的比例约为30%,而脾脏中约为50%;头肾和外周血中IgMλ B细胞与总Igλ B细胞的比例约为45%,而脾脏中约为25%。推测IgMλ B细胞为IgT B细胞。最后,我们检测到无乳链球菌感染后血清中针对无乳链球菌表面抗原-Sip的特异性抗体水平升高,表明小鼠抗罗非鱼Igλ单克隆抗体可用于检测尼罗罗非鱼免疫后的抗体水平,为评估罗非鱼疫苗的免疫效果奠定了基础。

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