Preparation of four monoclonal antibodies against human B7-1 and analysis of their biological characteristics / 中国免疫学杂志

ABSTRACT

Objective:To prepare the monoclonal antibody(mAb) against human B7 1 and analyse its biological characteristics.Methods:The B lymphocytes hybridization technique was applied by using XG7 B7 cell,a multiple myeloma(MM) cell line transfected with human B7 1 gene,as immunogen;the specificity and the antigen binding activity of mAbs were identified by flow cytometry and Western blot analysis;its biological effects on human PBTC and human B lymphoma cell line were examined by 3H TdR incroporation and annexin V satining.Results:Four mouse anti human B7 1(B7 1) hybridoma(1F11,3H8,6H2,7B10) were obtained.They secrete continuosly and steadily specific anti human B7 1(B7 1) mAb and their subclasses belong to IgG1 and IgM respectively;three of four mAbs could inhibit the proliferation of response cells(the human peripheral blood T lymphocytes),stimulated by costimulatory molecule B7 1.Furthermore,it was found that these mAbs induced the apoptosis of human B lymphoma cell line,Raji,which express naturally human B7 1 molecule,by using annexin V staining analysis after 24 hours of mAb treatment.Conclusion:Sucessefully obtained four mouse anti human B7 1 functional monoclonal antibodies,which have a potential value in anti allogenetic graft rejection and in the therapeutic approach of B lymphoma.