Anti-human kappa opioid receptor antibodies: characterization of site-directed neutralizing antibodies specific for a peptide kappa R(33-52) derived from the predicted amino terminal region of the human kappa receptor.
Site-directed polyclonal Abs specific for a synthetic peptide with sequence homology to the predicted N-terminal sequence of the human kappa opioid receptor [anti-kappa R-(33-52)] are capable of binding to normal human cells and cell lines expressing mRNA specific for the human kappa receptor. Flow cytometric analysis of 1) a neuronal cell line (NT2), 2) blood-derived CD14+ monocytes, 3) monocyte-like cell lines (U937 and THP 1), 4) blood-derived CD3+ T cells and a T cell line, and 5) human B cell lines bound anti-kappa R-(33-52) in a specific manner. Anti-kappa R-(33-52) was also found to specifically neutralize the immunosuppressive activities associated with the kappa R-selective agonist U50,488H. This antiserum was found to block U50,488H-mediated inhibition of 1) Staphylococcus aureus Cowen strain I-induced B and T lymphocyte proliferation, 2) PHA-induced T lymphocyte proliferation, and 3) S. aureus Cowen strain I-induced IgG production. However, this antiserum failed to neutralize mu R-selective agonist (Tyr-D-Ala-Gly-NMe-Phe-Gly-ol)-mediated suppression of IgG synthesis. Finally, the kappa R-selective antagonist nor-binaltorphimine hydrochloride inhibits the binding of anti-kappa R-(33-52) to the U937 cell line. These results suggest that anti-kappa R-(33-52) specifically interacts with the human kappa R molecule. Studies conducted with anti-kappa R-(33-52) indicated that this antiserum effectively blocked U50,488H-mediated immunosuppression, but by itself did not enhance or suppress lymphocyte activation. These data suggest that anti-kappa R-(33-52) 1) does not interact with the effector binding site of the receptor, but sterically interferes with U50,488H binding to the receptor; and/or 2) the antiserum interacts with a secondary binding site that is important for ligand binding, but may not be involved in signal transduction.
Buchner
R R
RR
Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Vogen
S M
SM
Fischer
W
W
Thoman
M L
ML
Sanderson
S D
SD
Morgan
E L
EL
D000328:Adult; D000595:Amino Acid Sequence; D019138:Antibodies, Blocking / Q000494:pharmacology; D000918:Antibody Specificity; D001483:Base Sequence; D001666:Binding Sites, Antibody; D002460:Cell Line; D005260:Female; D006801:Humans; D007106:Immune Sera / Q000096:biosynthesis / Q000737:chemistry* / Q000494:pharmacology*; D007074:Immunoglobulin G / Q000096:biosynthesis; D007166:Immunosuppressive Agents / Q000037:antagonists & inhibitors / Q000276:immunology; D019169:Jurkat Cells; D008213:Lymphocyte Activation; D016403:Lymphoma, Large B-Cell, Diffuse; D008297:Male; D008969:Molecular Sequence Data; D009000:Monocytes; D009447:Neuroblastoma; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*; D012333:RNA, Messenger / Q000096:biosynthesis; D017473:Receptors, Opioid, kappa / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D019138:Antibodies, Blocking; D007106:Immune Sera; D007074:Immunoglobulin G; D007166:Immunosuppressive Agents; D010446:Peptide Fragments; D012333:RNA, Messenger; D017473:Receptors, Opioid, kappa