title: Establishment of a cell model for screening antibody drugs against rheumatoid arthritis with ADCC and CDC.
abstract: TNFα played a dominant role in the development and progression of rheumatoid arthritis (RA). Clinical trials proved the efficacies of anti-TNFα agents for curing RA. However, most researchers were concentrating on their abilities of neutralizing TNFα, the potencies of different anti-TNFα agents varied a lot due to the antibody-dependent cell-mediated cytotoxicity (ADCC) or complement dependent cytotoxicity (CDC). For better understanding and differentiating the potentiality of various candidate anti-TNF reagents at the stage of new drug research and development, present study established a cell model expressing the transmembrane TNFα for usage in in vitro ADCC or CDC assay, meanwhile, the assay protocol described here could provide guidelines for screening macromolecular antibody drugs. A stable cell subline bearing transmembrane TNFα was first established by conventional transfection method, the expression of transmembrane TNFα was approved by flow cytometer, and the performance of the stable subline in ADCC and CDC assay was evaluated, using human peripheral blood mononuclear cells as effector cells, and Adalimumab as the anti-TNFα reagent. The stable cell subline demonstrated high level of surface expression of transmembrane TNFα, and Adalimumab exerted both ADCC and CDC effects on this cell model. In conclusion, the stable cell line we established in present research could be used in ADCC or CDC assay for screening antibody drugs, which would provide in-depth understanding of the potencies of candidate antibody drugs in addition to the traditional TNFα neutralizing assay.
authors.affiliation: Department of Reparative and Reconstructive Surgery of Orthopedics, Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430033, China.
authors.forename: Li
authors.initials: L
authors.lastname: Yan
authors.affiliation: Department of Reparative and Reconstructive Surgery of Orthopedics, Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430033, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Hu
authors.affiliation: Department of Reparative and Reconstructive Surgery of Orthopedics, Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430033, China.
authors.forename: Song
authors.initials: S
authors.lastname: Tu
authors.affiliation: Department of Reparative and Reconstructive Surgery of Orthopedics, Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430033, China.
authors.forename: Wen-Jun
authors.initials: WJ
authors.lastname: Cheng
authors.affiliation: Department of Reparative and Reconstructive Surgery of Orthopedics, Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430033, China.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Zheng
authors.affiliation: Department of Reparative and Reconstructive Surgery of Orthopedics, Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430033, China.
authors.forename: Jun-Wen
authors.initials: JW
authors.lastname: Wang
authors.affiliation: Department of Reparative and Reconstructive Surgery of Orthopedics, Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430033, China.
authors.forename: Wu-Sheng
authors.initials: WS
authors.lastname: Kan
authors.affiliation: Department of Reparative and Reconstructive Surgery of Orthopedics, Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430033, China.
authors.forename: Yi-Jun
authors.initials: YJ
authors.lastname: Ren
keywords: Transmembrane TNFα; antibody-dependent cell-mediated cytotoxicity; complement dependent cytotoxicity; rheumatoid arthritis