title: [Effect of CD80-scFv on recognition of CD80 molecule and proliferation of different tumor cells in vitro].
abstract: OBJECTIVE
  To prepare CD80-scFv in CHO cells and investigate  its effect on the recognition and proliferation of different tumor cells.
  
  
  METHODS
  The CD80-scFv was purified from culture supernatant without fetal calf serum (FCS) by IMAC affinity chromatography,  and then bound to CD80 molecule on the Daudi,  U251,  A375 and 8266 cells detected by flow cytometry. Different concentrations of CD80-scFv were added in the training system of different tumor cells,  and we analyzed the influence on the proliferation of these cells by MTT assay. With 8266 cells which expressed CD80 highly and were treated by mitomycin firstly as the APC,  and human PBMC as the effect cells,  we analyzed the influence of the CD80-scFv on the proliferation of PBMC by blocking the co-stimulatory signal.
  
  
  RESULTS
  The concentration of CD80-scFv purified from FCS was about 6.67 mg/L and the binding rate of CD80-scFv with Daudi,  U251,  A375,  8266 and U266 were 96.8%,  39.6%,  20.5%,  99.9% and 3.8%,  respectively. CD80-scFv suppressed the proliferation of Daudi and 8266 cells which expressed CD80 highly,  and the inhibition rate of Daudi and 8266 cells cultured with CD80-scFv (the final concentration was 25 μg/mL) was 24.04% and 24.16%,  respectively. Additionally,  the antibody suppressed the proliferation of PBMC by blocking the co-stimulatory signal mediated by CD80-CD28.
  
  
  CONCLUSIONS
  CD80-scFv can recognize CD80 molecule and suppress the proliferation of tumor cells which express CD80 highly.
authors.affiliation: Department of Immunology, College of Basic Medicine and Biosciences, Soochow University, Suzhou, China.
authors.forename: Li-jun
authors.initials: LJ
authors.lastname: Shen
authors.forename: Lian-hua
authors.initials: LH
authors.lastname: Han
authors.forename: Hua-ting
authors.initials: HT
authors.lastname: Zhu
authors.forename: Yong
authors.initials: Y
authors.lastname: Kong
authors.forename: Meng
authors.initials: M
authors.lastname: An
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Yong-jie
authors.initials: YJ
authors.lastname: Xia
authors.forename: Yu-hua
authors.initials: YH
authors.lastname: Qiu
mesh_terms: D000818:Animals; D018122:B7-1 Antigen / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D016466:CHO Cells; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D009369:Neoplasms / Q000276:immunology / Q000378:metabolism*; D011485:Protein Binding / Q000276:immunology; D057127:Single-Chain Antibodies / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D018122:B7-1 Antigen; D057127:Single-Chain Antibodies