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
| Autor: | R R Buchner, S M Vogen, W Fischer, M L Thoman, S D Sanderson, E L Morgan |
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| Rok vydání: | 1997 |
| Předmět: | |
| Zdroj: | The Journal of Immunology. 158:1670-1680 |
| ISSN: | 1550-6606 0022-1767 |
| DOI: | 10.4049/jimmunol.158.4.1670 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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