Identification of high affinity receptors for human monocyte chemoattractant protein-1 on human monocytes

Teizo Yoshimura, E. J. Leonard

Research output: Contribution to journalArticle

115 Citations (Scopus)

Abstract

The binding of human monocyte chemoattractant protein-1 (MCP-1) to human monocytes was studied. MCP-1 was radioiodinated with Iodo-beads (Pierce Chemical Co., Rockford, IL) without significant loss of biologic activity. 125I-MCP-1 binding to PBMC occurred within 5 min at 0°C and the binding was inhibited by unlabeled MCP-1 dose dependently but not by neutrophil attractant/activation protein-1 or FMLP. 125I-MCP-1 bound to monocytes; no significant binding to either neutrophils or lymphocytes was observed. Scatchard plot analysis indicated that monocytes had a minimum of 1700 ± 600 binding sites per cell with a K(d) of 1.9 ± 0.2 x 10-9 M. For analysis of binding by flow cytometry, MCP-1 was biotinylated. In contrast to radioiodination, biotinylation resulted in loss of activity; potency was 10-fold less, but the efficacy was retained. Detection by flow cytometry of bound biotinylated MCP-1 with avidin-FITC confirmed results obtained with 125I-MCP-1. Biotinylated MCP-1 bound to monocytes but not to lymphocytes; and the binding was inhibited by a 100-fold excess of unlabeled MCP-1.

Original languageEnglish
Pages (from-to)292-297
Number of pages6
JournalJournal of Immunology
Volume145
Issue number1
Publication statusPublished - 1990
Externally publishedYes

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Chemokine CCL2
Monocytes
Flow Cytometry
Lymphocytes
Biotinylation
human CCL2 protein
Neutrophil Activation
Protein Binding
Neutrophils
Binding Sites

ASJC Scopus subject areas

  • Immunology

Cite this

Identification of high affinity receptors for human monocyte chemoattractant protein-1 on human monocytes. / Yoshimura, Teizo; Leonard, E. J.

In: Journal of Immunology, Vol. 145, No. 1, 1990, p. 292-297.

Research output: Contribution to journalArticle

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