Functional analysis of TRAIL receptors using monoclonal antibodies.
Functional analysis of TRAIL receptors using monoclonal antibodies.
复制标题
使用单克隆抗体对 TRAIL 受体进行功能分析。
作者：
T. Griffith;C. Rauch;P. J. Smolak;Jennifer Y. Waugh;N. Boiani;D. Lynch;Craig A. Smith;R. Goodwin;M. Kubin
针对四种已知的tnf相关凋亡诱导配体（TRAIL）受体的细胞外结构域生成单克隆抗体，并在一组人类黑色素瘤细胞系上进行了测试。该单抗的特异性允许对诱导细胞凋亡的TRAIL受体（TRAIL- r1和-R2）与可能调节TRAIL介导的细胞凋亡的TRAIL受体（TRAIL- r3和-R4）进行精确评估。固定的抗TRAIL- r1或-R2单克隆抗体对TRAIL敏感的肿瘤细胞具有细胞毒性，而对重组TRAIL耐药的肿瘤细胞也对这些单克隆抗体耐药，只有在放线菌素d培养时才变得敏感。抗TRAIL- r1和-R2单克隆抗体诱导的死亡的特征是细胞内半胱天酶的激活。可被碳苄氧基-缬氨酸- α - asp （OMe）氟甲基酮（zVAD-fmk）和碳苄氧基- il - glu (OMe)-Thr-Asp （OMe）氟甲基酮（zIETD-fmk）阻断。在溶液中使用时，其中一种抗TRAIL- r2单克隆抗体能够阻断亮氨酸拉链-人TRAIL与TRAIL- r2表达细胞的结合，并阻止TRAIL诱导的细胞死亡，而两种抗TRAIL- r1单克隆抗体能够抑制亮氨酸拉链-人TRAIL与TRAIL- r1:Fc的结合。此外，使用阻断抗TRAIL-R2 mAb使我们能够证明通过TRAIL-R1或TRAIL-R2转导的信号是介导细胞死亡的必要和充分的。相反，TRAIL- r3或TRAIL- r4的表达似乎并不是决定这些肿瘤靶细胞对TRAIL作用的抗性或敏感性的重要因素。
mAbs were generated against the extracellular domain of the four known TNF-related apoptosis-inducing ligand (TRAIL) receptors and tested on a panel of human melanoma cell lines. The specificity of the mAb permitted a precise evaluation of the TRAIL receptors that induce apoptosis (TRAIL-R1 and -R2) compared with the TRAIL receptors that potentially regulate TRAIL-mediated apoptosis (TRAIL-R3 and -R4). Immobilized anti-TRAIL-R1 or -R2 mAbs were cytotoxic to TRAIL-sensitive tumor cells, whereas tumor cells resistant to recombinant TRAIL were also resistant to these mAbs and only became sensitive when cultured with actinomycin D. The anti-TRAIL-R1 and -R2 mAb-induced death was characterized by the activation of intracellular caspases, which could be blocked by carbobenzyloxy-Val-Ala-Asp (OMe) fluoromethyl ketone (zVAD-fmk) and carbobenzyloxy-Ile-Glu(OMe)-Thr-Asp (OMe) fluoromethyl ketone (zIETD-fmk). When used in solution, one of the anti-TRAIL-R2 mAbs was capable of blocking leucine zipper-human TRAIL binding to TRAIL-R2-expressing cells and prevented TRAIL-induced death of these cells, whereas two of the anti-TRAIL-R1 mAbs could inhibit leucine zipper-human TRAIL binding to TRAIL-R1:Fc. Furthermore, use of the blocking anti-TRAIL-R2 mAb allowed us to demonstrate that the signals transduced through either TRAIL-R1 or TRAIL-R2 were necessary and sufficient to mediate cell death. In contrast, the expression of TRAIL-R3 or TRAIL-R4 did not appear to be a significant factor in determining the resistance or sensitivity of these tumor target cells to the effects of TRAIL.
影响因子：
56.9
作者：
CHENG, JH;ZHOU, T;MOUNTZ, JD
通讯作者：
MOUNTZ, JD