An antagonistic monoclonal antibody (B-N6) specific for the human neurotensin receptor-1

Neuropeptides. 1998 Jun;32(3):247-56. doi: 10.1016/s0143-4179(98)90044-0.

Abstract

The neuropeptide neurotensin (NT) interacts with two types of human receptors (hNTR) termed hNTR-1 and hNTR-2. This study describes a monoclonal antibody (MAb) specific for hNTR-1, B-N6. This MAb binds specifically to hNTR-1, but not to hNTR-2 transfected CHO cells. B-N6 and NT display a reciprocal competition and react in a similar way to trypsin, suggesting that the B-N6 epitope is at or close to the NT binding site on the third extracellular loop. Unlike B-N6, NT induces hNTR-1 internalization. Although neither NT-FITC nor B-N6 binding was detected by flow cytometry on different human cells, specific mRNA expression for hNTR-1 was detected in these cells. In CHO cells expressing hNTR-1 and a luciferase gene coupled to the krox24 reporter, B-N6 and the antagonist SR 48692 inhibited NT-induced intracellular activation of krox24 in a dose-dependent manner. From these results it is concluded that B-N6 is an antagonistic anti-hNTR-1 MAb.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / metabolism
  • Antibodies, Monoclonal / pharmacology*
  • Antibody Specificity
  • Binding Sites, Antibody
  • CHO Cells
  • Cell Line
  • Cricetinae
  • DNA-Binding Proteins / genetics
  • Early Growth Response Protein 1
  • Flow Cytometry
  • Humans
  • Immediate-Early Proteins*
  • Luciferases / genetics
  • Mice
  • Mice, Inbred BALB C
  • Neurotensin / metabolism
  • Neurotensin / pharmacology
  • RNA, Messenger / analysis
  • Receptors, Neurotensin / antagonists & inhibitors*
  • Receptors, Neurotensin / genetics
  • Receptors, Neurotensin / metabolism
  • Recombinant Proteins
  • Transcription Factors / genetics
  • Transfection
  • Trypsin / metabolism
  • Tumor Cells, Cultured

Substances

  • Antibodies, Monoclonal
  • DNA-Binding Proteins
  • EGR1 protein, human
  • Early Growth Response Protein 1
  • Egr1 protein, mouse
  • Immediate-Early Proteins
  • RNA, Messenger
  • Receptors, Neurotensin
  • Recombinant Proteins
  • Transcription Factors
  • Neurotensin
  • Luciferases
  • Trypsin