Military Medical Sciences 2014;(8):617-620,625

doi:10.7644/j.issn.1674-9960.2014.08.012

Biological activity and application study of a monoclonal antibody against human Tim-3

Chunmei HOU ; Xiaoling LANG ; Yan XIAO ; Mingke ZHENG ; Xingwei JIANG ; He XIAO ; Guojiang CHEN ; Renxi WANG ; Jiannan FENG ; Gencheng HAN ; Beifen SHEN ; Yan LI

Keywords

Tim-3; antibodies,monoclonal; L3D; U937 cells; inflammatory factors; sepsis

Country

China

Language

Chinese

Abstract

Objective To develop a human Tim-3 specific monoclonal antibody and evaluate its biological activity and possible use in clinical diseases associated with dysregulated Tim-3 expression .Methods The BALB/c mice were immu-nized by conventional method, and positive clones were used to develop anti-human Tim-3 antibody, the binding and neutralization activities of which in vitro and in vivo were investigated.Results ①A monoclonal antibody (clone L3D) which could specifically bind to human Tim-3 protein in ELISA assay was obtained and the subtype of the monoclonal antibody was IgG2a .②Flow cytometry indicated that the monoclonal antibody could bind to Tim-3 expressed in human U937 cells.This antibody also showed a cross activity to mice′Tim-3.③The monoclonal antibody inhibited the apoptosis of THP1 cells induced by Gal-9, the ligand of Tim-3.④Injection of Tim-3 antibody exacerbated sepsis in mice as marked by the decreased survival rate and increased expression of pro-inflammatory cytokines .Conclusion An anti-human Tim-3 monoclonal antibody is successfully obtained.The excellent binding and neutralization activities of this antibody enable it to be widely used in clinical diseases associated with deregulated Tim-3 expression .