摘要
目的:制备抗人RANTES分子单克隆抗体(mAb)并进行初步鉴定,为研究RANTES分子的组织分布和功能提供实验手段。方法:应用淋巴细胞杂交瘤技术,制备小鼠源性抗人RANTESmAb。用ELISA法鉴定腹水mAb的效价。用Q FastFlow阴离子交换柱纯化mAb。用Westernblot鉴定mAb的抗原结合活性。用免疫组化染色法,对RANTES分子在进行小肠移植术的大鼠移植肠组织中的分布进行鉴定。结果:获得4株分泌抗RANTESmAb的杂交瘤细胞株。间接ELISA法测定腹水mAb的效价均达1×10-6,3株mAb为IgG1亚类(κ),1株为IgG2b(κ)。Westernblot的结果显示,3株mAb与人RANTES均有良好的结合活性。用3株mAb进行免疫组化染色的结果显示,RANTES分子在进行小肠移植术的大鼠小肠腺上皮细胞胞质中呈高表达。结论:获得4株能特异性识别天然RANTES分子的mAb。大鼠小肠移植后其肠上皮细胞中可高水平地表达RANTES。
AIM: To prepare monoclonal antibodies (against) human RANTES molecule and identify the expression of RANTES in rat small intestine after small bowel transplantation. METHODS: Murine mAbs were prepared by B lymphocyte hybridoma technique. The expression of RANTES in rat small intestine after small bowel transplantation was detected by immunohistochemistry. RESULTS: Four hybridoma cell lines secreting monoclonal antibodies to human RANTES, FMU-RANTES 1, FMU-RANTES 2, FMU-RANTES 3 and FMU-RANTES 4, were established. The titers of a scetic mAbs reached to 1×10^(-6) and the Ig subclass of FMU-RANTES 1, FMU-RANTES 3 and FMU-RANTES 4 was IgG1(κ) and that of FMU-RANTES 2 was IgG2b(κ). Among these mAbs, FMU-RANTES 1, FMU-RANTES 2 and FMU-RANTES 3 could bind human RANTES protein in Western bolt. FMU-RANTES 1, FMU-RANTES 2 and FMU-RANTES 4 could be used in immunohistochemistry staining. Rat RANTES molecule could be detected in the cyto plasm of epithelial cells in rat small intestine after small bowel transplantation. CONCLUSION: Four mAbs against RANTES molecule were prepared, which can provide a useful tool in research on the structure and function of RANTES molecule. High expression of RANTES may be involved in the rejection of allogeneic graft.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2005年第3期322-324,327,共4页
Chinese Journal of Cellular and Molecular Immunology
