TRAIL(TNF related apoptosis inducing ligand)是新发现的肿瘤坏死因子家族(TNF)成员,能够诱导多种肿瘤细胞凋亡,其中受体决定着TRAIL的生物学功能,在TRAIL诱导的细胞凋亡中发挥着重要作用,随着研究的不断深入,发现其受体的结构组成与...TRAIL(TNF related apoptosis inducing ligand)是新发现的肿瘤坏死因子家族(TNF)成员,能够诱导多种肿瘤细胞凋亡,其中受体决定着TRAIL的生物学功能,在TRAIL诱导的细胞凋亡中发挥着重要作用,随着研究的不断深入,发现其受体的结构组成与功能比原先所认识的更为复杂,并且在组成结构与功能上存在不一致性,提示TRAIL受体对TRAIL功能的调节具有系统性,尚有许多待认识和研究的地方,本文仅对TRAIL受体的研究新进展进行综述。展开更多
Objective To establish hybridomas that produce anti-death receptor-5 (DR5) monoclonal antibodies (mAbs) and check the surface expression of DR5 (sDR5) on cell lines.Methods The cDNA of human DR5 was cloned into pGAPZ...Objective To establish hybridomas that produce anti-death receptor-5 (DR5) monoclonal antibodies (mAbs) and check the surface expression of DR5 (sDR5) on cell lines.Methods The cDNA of human DR5 was cloned into pGAPZα. Recombinant Pichia pastoris clones generated via homologous recombination secreted high levels of sDR5. The sDR5 was purified using a nickel ion column. BALB/c mice were immunized with sDR5 and spleen cells were fused with the SP2/0-Ag 14. Monoclonal antibodies were tested by ELISA for their abilities binding to sDR5 and by flow cytometry for the reactivities to surface DR5 of Jurkat cells. Surface expression of the TRAIL receptor was determined by flow cytometric analysis measuring the binding of anti-DR5 mAb.Results Isotypes of mAbs were determined to be IgG, and IgM, all of which were reactive to sDR5 as observed through ELISA. It was discovered using flow cytometry that only IgG was able to bind to DR5 on the plasma membrane of Jurkat cells. sDR5 was found to completely inhibit anti-DR5 mAb binding to Jurkat cells. Approximately 95% of Jurkat cells, 98% SW480, 99% U937, 100% U87, 86% HCT116, 64% HL-60, 47% HeLa and 13% K562 cells express membrane DR5.Conclusions These results demonstrate that anti-DR5 mAb is able to specifically bind to DR5 and that DR5 is expressed at high levels on Jurkat, SW480, U87, U937 and HCT116 cell lines, and at medium levels on HL-60 and HeLa cell lines. The expression of DR5 on K562 cell line is low.展开更多
文摘TRAIL(TNF related apoptosis inducing ligand)是新发现的肿瘤坏死因子家族(TNF)成员,能够诱导多种肿瘤细胞凋亡,其中受体决定着TRAIL的生物学功能,在TRAIL诱导的细胞凋亡中发挥着重要作用,随着研究的不断深入,发现其受体的结构组成与功能比原先所认识的更为复杂,并且在组成结构与功能上存在不一致性,提示TRAIL受体对TRAIL功能的调节具有系统性,尚有许多待认识和研究的地方,本文仅对TRAIL受体的研究新进展进行综述。
基金supported in part by grants HNIFOS 0321001800 and HNIFMSTS 2002-119.
文摘Objective To establish hybridomas that produce anti-death receptor-5 (DR5) monoclonal antibodies (mAbs) and check the surface expression of DR5 (sDR5) on cell lines.Methods The cDNA of human DR5 was cloned into pGAPZα. Recombinant Pichia pastoris clones generated via homologous recombination secreted high levels of sDR5. The sDR5 was purified using a nickel ion column. BALB/c mice were immunized with sDR5 and spleen cells were fused with the SP2/0-Ag 14. Monoclonal antibodies were tested by ELISA for their abilities binding to sDR5 and by flow cytometry for the reactivities to surface DR5 of Jurkat cells. Surface expression of the TRAIL receptor was determined by flow cytometric analysis measuring the binding of anti-DR5 mAb.Results Isotypes of mAbs were determined to be IgG, and IgM, all of which were reactive to sDR5 as observed through ELISA. It was discovered using flow cytometry that only IgG was able to bind to DR5 on the plasma membrane of Jurkat cells. sDR5 was found to completely inhibit anti-DR5 mAb binding to Jurkat cells. Approximately 95% of Jurkat cells, 98% SW480, 99% U937, 100% U87, 86% HCT116, 64% HL-60, 47% HeLa and 13% K562 cells express membrane DR5.Conclusions These results demonstrate that anti-DR5 mAb is able to specifically bind to DR5 and that DR5 is expressed at high levels on Jurkat, SW480, U87, U937 and HCT116 cell lines, and at medium levels on HL-60 and HeLa cell lines. The expression of DR5 on K562 cell line is low.
