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 language | English |
|---|---|
| Pages (from-to) | 292-297 |
| Number of pages | 6 |
| Journal | Journal of Immunology |
| Volume | 145 |
| Issue number | 1 |
| Publication status | Published - 1990 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Identification of high affinity receptors for human monocyte chemoattractant protein-1 on human monocytes
AU - Yoshimura, Teizo
AU - Leonard, E. J.
PY - 1990
Y1 - 1990
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0025307165&partnerID=8YFLogxK
M3 - Article
C2 - 2162890
AN - SCOPUS:0025307165
VL - 145
SP - 292
EP - 297
JO - Journal of Immunology
JF - Journal of Immunology
SN - 0022-1767
IS - 1
ER -