抗草鱼c来d4l.docx

中文摘要 中文摘要 抗草鱼CD4L-2单克隆抗体的制备及其应用研究 低的顺序为胸腺、鳃、肝脏、肠道、脾脏、头肾、体肾、血液，其中，胸腺的CD4L-2+ 细胞含量极显著地高于血液与体肾(pO．01)，鳃、肝脏和肠道中CD4L-2+细胞含 量显著高于血液与体‘肾(p0．05)。用PHA刺激48 h后，各免疫相关组织中CD4L-2+ 细胞比例发生了明显变化，其中，头肾、脾脏、肝脏与肠道中CD4L-2+细胞比例 显著升高，而胸腺中CD4L-2+细胞比例有所下降。激光共聚焦分析显示，肠粘膜 中嗜酸性细胞、嗜中性细胞、单核细胞、淋巴细胞等均为CD4L-2+细胞。 结论本研究克隆了草鱼CD4L-2基因，制备并鉴定了抗草鱼CD4L-2单克隆 抗体，探明了草鱼CD4L-2+细胞在相关组织中的分布特征，初步明确了草鱼肠粘 膜中表达CD4L-2+的细胞类型。鉴于CD4+T细胞在获得性免疫系统中的作用，抗 草鱼CD4L-2单克隆抗体的获得，为鱼类T细胞亚群的分类及功能研究奠定了基 础。 关键词：草鱼；CD4L-2；单克隆抗体；CD4L-2+细胞 作 者：柳昭君 指导教师：宋学宏 Ⅱ 万方数据 抗草鱼CD4L-2单克隆抗体的制各及其应用研究 抗草鱼CD4L-2单克隆抗体的制各及其应用研究 英文摘要 Preparation and application of monoclonal antibody against grass carp C D4L-2 Abstract Objective：CD4 molecule is a single chain transmembrane glycoprotein found on the surface of lymphocytes and all immunoglobulin family membrane-anchored protein， which plays an important role in immune responses by binding tO MHC—II molecules and involving in TCR signaling in Th cells．The grass carp(Ctenopharyngodon idella) is a major aquaculture species in China，and is also a representative species of teleost， the major primitive vertebrate taxa．Investigation on the immune system in fish will not only deepen Our understanding of the diversity and evolution of vertebrate immune system，but provide theoretical basis for the prevention and control of aquatic diseases as well．The aim of this study is to prepare monoclonal antibody against the grass carp CD4，and SOrt out CD4+cells，and then lay a foundation for classification of T cell subsets and for their functional analysis． Methods：The specific primers were designed based on partial sequence from RNA-seq data，and then the grass carp CD4L-2 gene was isolated by using PCR technique．The expression patterns of CD4L-2 were examined by qPCR assay．Highly antigenic CD4L-2 extracellular protein was obtained by prokaryotic expression，