Functional differences between monocyte chemotactic protein-1 receptor A and monocyte chemotactic protein-1 receptor B expressed in a Jurkat T cell

J Immunol. 2000 Nov 1;165(9):4877-83. doi: 10.4049/jimmunol.165.9.4877.

Abstract

The monocyte chemotactic protein-1 (MCP-1) receptor (MCP-1R) is expressed on monocytes, a subpopulation of memory T lymphocytes, and basophils. Two alternatively spliced forms of MCP-1R, CCR2A and CCR2B, exist and differ only in their carboxyl-terminal tails. To determine whether CCR2A and CCR2B receptors function similarly, Jurkat T cells were stably transfected with plasmids encoding the human CCR2A or CCR2B gene. Nanomolar concentrations of MCP-1 induced chemotaxis in the CCR2B transfectants that express high, intermediate, and low levels of MCP-1R. Peak chemotactic activity was shifted to the right as receptor number decreased. Five-fold more MCP-1 was required to initiate chemotaxis of the CCR2A low transfectant, but the peak of chemotaxis was similar for the CCR2A and CCR2B transfectants expressing similar numbers of receptors. MCP-1-induced chemotaxis was sensitive to pertussis toxin, implying that both CCR2A and CCR2B are G(i)alpha protein coupled. MCP-1 induced a transient Ca(2+) flux in the CCR2B transfectant that was partially sensitive to pertussis toxin. In contrast, MCP-1 did not induce Ca(2+) flux in the CCR2A transfectant. Since MCP-1 can stimulate chemotaxis of the CCR2A transfectant without inducing Ca(2+) mobilization, Ca(2+) flux may not be required for MCP-1-induced chemotaxis in the Jurkat transfectants. These results indicate that functional differences exist between the CCR2A and CCR2B transfectants that can be attributed solely to differences in the carboxyl-terminal tail.

Publication types

  • Comparative Study

MeSH terms

  • Calcium / antagonists & inhibitors
  • Calcium / metabolism
  • Calcium Signaling / genetics
  • Calcium Signaling / immunology
  • Chemokine CCL2 / antagonists & inhibitors
  • Chemokine CCL2 / genetics
  • Chemokine CCL2 / metabolism*
  • Chemokine CCL2 / pharmacology
  • Chemotaxis, Leukocyte / genetics
  • Chemotaxis, Leukocyte / immunology
  • Guanosine 5'-O-(3-Thiotriphosphate) / pharmacology
  • Humans
  • Iodine Radioisotopes
  • Jurkat Cells / immunology*
  • Jurkat Cells / metabolism*
  • Pertussis Toxin
  • Protein Binding / genetics
  • Protein Binding / immunology
  • Receptors, CCR2
  • Receptors, Chemokine / antagonists & inhibitors
  • Receptors, Chemokine / biosynthesis*
  • Receptors, Chemokine / genetics
  • Receptors, Chemokine / physiology
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism*
  • Transfection
  • Virulence Factors, Bordetella / pharmacology

Substances

  • CCR2 protein, human
  • Chemokine CCL2
  • Iodine Radioisotopes
  • Receptors, CCR2
  • Receptors, Chemokine
  • Virulence Factors, Bordetella
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • Pertussis Toxin
  • Calcium