Journal article
Serologic analysis of the mouse beta chemokine JE/monocyte chemoattractant protein-1
The Journal of immunology (1950), Vol.153(8), pp.3708-3716
10/15/1994
DOI: 10.4049/jimmunol.153.8.3708
PMID: 7523503
Abstract
Mouse monocyte chemoattractant protein-1 (MCP-1), previously termed JE, is a member of the beta chemokine gene family and a homologue of the human monocyte chemoattractant protein, MCP-1. Mouse rMCP-1 was used to immunize hamsters for the production of mAb. Seven mouse MCP-1-specific mAbs were characterized: two of these mAbs cross-reacted with the human MCP-1, as determined by ELISA. A sensitive and specific capture ELISA for MCP-1 quantitation, which allowed measurement of mouse MCP-1 levels in supernatants from cells stimulated with inflammatory agents, was developed. LPS-stimulated astrocytes produce the highest levels of MCP-1 (80 ng/ml); macrophages and mesangial cells produce lower levels of MCP-1 (2 to 14 ng/ml) after LPS stimulation. IL-1 and TNF-alpha stimulation also can induce low levels of MCP-1 production. Western blot analysis demonstrated that the predominant native form of mouse MCP-1 is a 30-kDa glycoprotein. Two mAbs (2H5 and 6C7) demonstrated dose-dependent neutralization of mouse MCP-1 chemotactic activity. To localize the epitope recognized by one of these neutralizing Abs, the mAb was used to bind a series of genetically engineered truncated variants of human MCP-1. The C-terminal residues 62 to 67 on human MCP-1 molecules seem to be critical to express the epitope recognized by the neutralizing 2H5 anti-MCP-1 mAb. However, multiple sites on the MCP-1 molecule seem to be critical for bioactivity. Thus, these Ab reagents provide a useful tool to explore the biology of the mouse MCP-1 beta chemokine.
Details
- Title: Subtitle
- Serologic analysis of the mouse beta chemokine JE/monocyte chemoattractant protein-1
- Creators
- Yi LuoJoseph LaningMasaharu HayashiPaul R HancockBarrett RollinsMartin E Dorf
- Resource Type
- Journal article
- Publication Details
- The Journal of immunology (1950), Vol.153(8), pp.3708-3716
- DOI
- 10.4049/jimmunol.153.8.3708
- PMID
- 7523503
- ISSN
- 0022-1767
- eISSN
- 1550-6606
- Language
- English
- Date published
- 10/15/1994
- Academic Unit
- Urology
- Record Identifier
- 9984319979802771
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