Characterization of the CCR2 chemokine receptor: functional CCR2 receptor expression in B cells.
We have derived anti-human CCR2-specific mAbs by immunization with synthetic peptides corresponding to CCR2 sequences presumably involved in the interaction with its ligand(s). The characterization of these mAbs includes the ability to recognize the CCR2 receptor specifically, as well as the function based on their ability to promote Ca2+ influx or to block MCP-1-induced Ca2+ influx and chemotaxis. One mAb (MCP-1 R02) that is directed to the NH2 terminal domain of the CCR2 receptor has MCP-1 agonist activity, and two that recognize the third extracellular domain (MCP-1R04 and MCP-1 R05) have MCP-1 antagonist activity. We analyzed the presence of CCR2 in several PBL and tonsil-derived leukocyte populations and found expression of this receptor in monocytes, activated T cells, and, surprisingly, in B cells. CCR2 receptor expression in B cells was further corroborated in Southern blot using CCR2-specific probes. Moreover, both MCP-1 and the agonist mAb trigger specific B cell migration via a PTX-sensitive mechanism, indicating the presence of a functional CCR2 receptor in these cells.
Frade
J M
JM
Department of Immunology and Oncology, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Cientificas, Campus Cantoblanco, Universidad Autónoma, Madrid, Spain.
Mellado
M
M
del Real
G
G
Gutierrez-Ramos
J C
JC
Lind
P
P
Martinez-A
C
C
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001402:B-Lymphocytes / Q000378:metabolism*; D002460:Cell Line; D018932:Chemokine CCL2 / Q000276:immunology; D006801:Humans; D054390:Receptors, CCR2; D019707:Receptors, Chemokine / Q000276:immunology / Q000378:metabolism*; D010566:Virulence Factors, Bordetella / Q000276:immunology
D000911:Antibodies, Monoclonal; C515334:CCR2 protein, human; D018932:Chemokine CCL2; D054390:Receptors, CCR2; D019707:Receptors, Chemokine; D010566:Virulence Factors, Bordetella