Preparation and characterization of a set of monoclonal antibodies to TRAIL and TRAIL receptors DR4, DR5, DcR1, and DcR2.
The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo 2L) is a novel cytotoxic ligand belonging to TNF superfamily. Among TRAIL receptors, death receptor 4 (DR4) and DR5 containing death domain (DD) in their cytoplasmic region mediate apoptosis-signaling upon TRAIL binding, while decoy receptor 1 (DcR1) and DcR2 with a truncated or non-functional DD play "decoy" role. The interaction of TRAIL and TRAIL receptors plays important roles both in immunoregulation and immune pathogenesis of some diseases. In this study, we raised hybridomas secreting monoclonal antibodies against TRAIL (FMU1.1, 1.2, 1.3), DR4 (FMU1.4), DR5 (FMU1.5, 1.6), DcR1 (FMU1.7) and DcR2 (FMU1.8, 1.9). These MAbs could be used for fluorescent staining and flow cytometry (FCM) analysis as well as immunohistochemistry (IHC). Moreover, FMU1.1, 1.3, 1.4 and 1.5 could be used as coating antibodies paring corresponding polyclonal antibodies to develop sandwich ELISAs to quantitate the soluble TRAIL (sTRAIL), sDR4 or sDR5 in serum samples respectively. In addition, cross-linking of DR4/DR5 by FMU1.4 or FMU1.5 MAbs could induce apoptosis of some DR4/DR5-expressing tumor cells. Thus, this set of monoclonal antibodies against TRAIL or TRAIL receptors may be useful in expression phenotypic and functional study of TRAIL and TRAIL receptors.
Liu
Xue-Song
XS
Department of Immunology, Fourth Military Medical University, Xi'an 710032, China.
Zhu
Yong
Y
Han
Wei-Ning
WN
Li
Ying-Na
YN
Chen
Li-Hua
LH
Jia
Wei
W
Song
Chao-Jun
CJ
Liu
Fei
F
Yang
Kun
K
Li
Qi
Q
Jin
Bo-Quan
BQ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D051017:Apoptosis Regulatory Proteins; D005260:Female; D058851:GPI-Linked Proteins; D006801:Humans; D006825:Hybridomas; D008562:Membrane Glycoproteins / Q000032:analysis / Q000276:immunology*; D008565:Membrane Proteins / Q000032:analysis / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D018124:Receptors, Tumor Necrosis Factor / Q000032:analysis / Q000276:immunology*; D053320:Receptors, Tumor Necrosis Factor, Member 10c; D053221:TNF-Related Apoptosis-Inducing Ligand; D053319:Tumor Necrosis Factor Decoy Receptors; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D051017:Apoptosis Regulatory Proteins; D058851:GPI-Linked Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D018124:Receptors, Tumor Necrosis Factor; D053320:Receptors, Tumor Necrosis Factor, Member 10c; D053221:TNF-Related Apoptosis-Inducing Ligand; C105190:TNFRSF10A protein, human; C505639:TNFRSF10B protein, human; C108661:TNFRSF10C protein, human; C451501:TNFRSF10D protein, human; C505653:TNFSF10 protein, human; C505640:Tnfrsf10b protein, mouse; C505654:Tnfsf10 protein, mouse; D053319:Tumor Necrosis Factor Decoy Receptors; D014409:Tumor Necrosis Factor-alpha