医学分子生物学杂志2020，Vol.17Issue(1)：52-56,5.DOI:10.3870/j.issn.1672-8009.2019.01.009
抗TIGIT单克隆抗体的制备及活性鉴定
Preparation and Activity Identification of Anti-TIGIT Monoclonal An-tibody
摘要
Abstract
Objective A monoclonal antibody against TIGIT was prepared and its effects on cardiac allograft tolerance was examined. Methods Mice were immunized with human TIGIT-GST recombinant protein and monoclonal antibodies were prepared.Antibody specificity assay was used to detect the binding ability of antibodies against TIGIT.The effect of TIGIT antibody on the killing of P815 by NK cells was examined by using a redirected killing assay.The effect of TIGIT ac-tivating antibodies on the activity of CTLs and CD8 Treg cells was examined by enzyme absorption assay.Results A non-activated anti-TIGIT antibody 7M13 and an activated anti-TIGIT antibody 4D9 were successfully prepared.Antibody specificity experiments showed that both antibodies 7M13 and 4D9 bound to TIGIT in humans and mice.The retargeting killing experiment showed that the an-tibody 7M13 did not significantly inhibit the killing effect of NK cells on P815 cells, indicating that 7M13 can bind to TIGIT without activating TIGIT while 4D9 can significantly inhibit the killing func-tion of NK cells.The isolation of CTLs and CD8 Treg from 37 heart transplant patients, TIGIT acti- vating antibody treatment had no significant effects on cell viability, but significantly inhibited the proliferation of CTLs, but exerted no significant effect on the proliferation of CD8 Treg cells.Conclusion The non-activated anti-TIGIT antibody 7M13 and the activated anti-TIGIT anti-body 4D9 were successfully prepared, respectively, and 4D9 could significantly inhibit the killing function of NK cells.关键词
心脏移植/ TIGHT (T cell immunoglobulin and ITIM domain)/ 单克隆抗体/ NK细胞Key words
hearttransplantation/TcellimmunoglobulinandITIMdomain/monoclonalanti-body/NKcells分类
医药卫生引用本文复制引用
李国华,程伟华,康建军,王航辉,姜丽..抗TIGIT单克隆抗体的制备及活性鉴定[J].医学分子生物学杂志,2020,17(1):52-56,5.基金项目
资助项目: 延安市科学技术研究发展计划项目 (No.2018ks-22) This work was supported by a grant from the Science and Technology Research Program of Yan’an, China (No.2018ks-22) （No.2018ks-22）
