![]() ![]() The cells were fixed and reacted with FITC conjugated antimouse IgG. The antibody (4F2) was added to 293T cells transfected with pCAGGS-hSTEAP4 and the cells continued to be cultured for 30 minutes. (f) Anti-hSTEAP4 monoclonal antibody bound to hSTEAP4 proteins on cell surface. (e) hSTEAP4 proteins detected by anti-hSTEAP4 monoclonal antibody (4F2 IgG2a) in 293T cells transfected with pCAGGS-hSTEAP4. ![]() (d) hSTEAP4 expression in androgen stimulated LNCaP cells using immunofluorescent staining. For androgen treatment, LNCaP cells were cultured with 5-DHT (10-9 M) for 24 hours. (c) Detection of hSTEAP4 expression in LNCaP cells using flow cytometry analysis. The cells were reacted with the indicated serum and stained with PE conjugated antimouse IgG. ![]() The lower panels show 293T cells transfected with pCAGGS-hSTEAP4-IRES-GFP. The upper panels show 293T cells transfected with pMRX-hSTEAP1-IRES-GFP. (b) Specific binding of anti-STEAP antibody in flow cytometry analysis. 293T cells transiently transfected with pMRX-hSTEAP4-IRES-GFP were stained with sera, followed by PE conjugated antimouse IgG. (a) Flow cytometry analysis to detect anti-hSTEAP4 in sera from immunized mice. Our results suggest that DC immunization is an effective method to produce antibodies reactive to extracellular regions of plasma membrane proteins with multiple-transmembrane domains, and may be useful to develop antibody mediated antitumor therapies. The immunized mice produced significant amounts of antibodies against these proteins. Mice were immunized with these DCs and followed by repeated booster immunization with plasmids expressing each protein. DCs were transduced with genes encoding the human six transmembrane epithelial antigen of prostate 1 (STEAP1), STEAP4, and seven transmembrane prostate specific G-protein coupled receptor (PSGR). In order to develop methods to generate antibodies reactive to the extracellular domains of multipass plasma membrane proteins specifically expressed in malignant tumors, we examined the use of dendritic cells (DCs) for immunization. Antibody mediated therapeutic strategies against human malignant tumors have been widely authorized and clinically applied to cancer patients. ![]()
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |