[Preparation and characterization of two novel functional monoclonal antibodies against human OX40L].
OBJECTIVE
To prepare novel anti-OX40L functional monoclonal antibodies and characterize their distinct biological functions.


METHODS
Routine immunization of BALB/c mice by a tansfected cell line L929/OX40L expressing high level of OX40L as antigens. Then fuse the immunized spleen with Sp2/0, a kind of myloma, and screen the positive clones by FCS with L929/OX40L as a positive control.After acquisition of the hybidomas secreting anti-OX40L mAb, investigation of their biological activities by Western blot, rapid isotyping analysis, karyotype analysis, competitive inhibition test, indirect immunofluorescence and MTT incorporation assay were followed.


RESULTS
Obtain two stable hybridomas, 4D6 and 5C2, which could continuously secret specific anti-OX40L monoclonal antibodies. The following biological activity studies showed that these monoclonal antibodies could both recognize the natural OX40L expressed on the mature DC, especially the OX40L on the several leukemia cell lines, such as Jurkat, SHI-1, U937 etc. Furthermore, they could also suppress the proliferation of SHI-1 in vitro.


CONCLUSIONS
Two hybridomas secreting anti-OX40L monoclonal antibodies continuously and steadily have been established. These monoclonal antibodies could specifically recognize human OX40L and effect the proliferation of leukemia cell line SHI-1 through OX40/OX40L costimulatory signals.
Wu
Hong-ya
HY
Institute of Medical Biotechnology, Soochow University, Suzhou 215007, China. wuhongya000@sina.com
Wang
Qin
Q
Chen
Yong-jing
YJ
Xie
Fang
F
Mao
Yi-xiang
YX
Liu
Gao-qin
GQ
Zhang
Xue-guang
XG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000915:Antibody Affinity; D000918:Antibody Specificity; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007938:Leukemia / Q000188:drug therapy; D051379:Mice; D008807:Mice, Inbred BALB C; D053278:OX40 Ligand / Q000276:immunology*
D000911:Antibodies, Monoclonal; D053278:OX40 Ligand; C089251:TNFSF4 protein, human