Journal article
Characterization of Monoclonal Antibodies to Human Soluble MD-2 Protein
Hybridoma (2005), Vol.25(6), pp.349-357
12/2006
DOI: 10.1089/hyb.2006.25.349
PMID: 17203997
Abstract
Toll-like receptors (TLRs) are mammalian innate immune recognition receptors that are activated by pathogen associated molecular patterns (PAMPs). TLR4 is the signaling molecule of the lipopolysaccharide (LPS) receptor complex. TLR4 associates with its adapter molecule, MD-2, which is absolutely required for LPSinduced activation of TLR4. MD-2 exists as a cell surface protein in association with TLR4 and as secreted forms consisting of MD-2 monomers and multimers. To facilitate the studies of MD-2 distribution, abundance, and function, we produced monoclonal antibodies (MAbs) to baculovirally expressed soluble MD-2 (sMD-2). Eleven MAbs were characterized by enzyme-linked immunosorbent assay (ELISA) with soluble TLR4/MD-2 complex (sTLR4/MD-2) and sMD-2, Western blotting against sMD-2 monomer and multimers, and inhibition of direct LPS binding to sMD-2. Four MAbs preferentially recognized mainly MD-2 oligomers, not monomers, as judged by Western blotting and ELISA. Anti-MD-2 MAbs useful for indirect immunofluorescent staining of cells expressing TLR4 and MD-2 were identified. One MAb that recognized all forms of MD-2 was used in an ELISA to measure sMD-2 in normal human sera as well as sera from intensive care patients with and without sepsis. Serum levels of sMD-2 were undetectable or very low in normal and in nonsepsis patients but significantly (p < 0.05) increased in sepsis patients. These MAbs should therefore be very useful new tools for studies of MD-2 expression and function in health and disease.
Details
- Title: Subtitle
- Characterization of Monoclonal Antibodies to Human Soluble MD-2 Protein
- Creators
- Suganya Viriyakosol - Veterans Medical Research Foundation, San Diego, California., Department of Pathology and Medicine, University of California, San Diego, CaliforniaPaul B Mccray - Department of Pediatrics, The Inflammation Program, Veterans Administration Medical Center, Iowa City, IowaMark E Ashbaugh - Veterans Medical Research Foundation, San Diego, CaliforniaJayne Chu - Veterans Medical Research Foundation, San Diego, CaliforniaHong Peng Jia - Department of Pediatrics, The Inflammation Program, Veterans Administration Medical Center, Iowa City, IowaJerold Weiss - Department of Internal Medicine, Carver College of Medicine, University of Iowa; The Inflammation Program, Veterans Administration Medical Center, Iowa City, IowaTheo N Kirkland - Veterans Administration San Diego Healthcare System, San Diego, California., Department of Pathology and Medicine, University of California, San Diego, California
- Resource Type
- Journal article
- Publication Details
- Hybridoma (2005), Vol.25(6), pp.349-357
- DOI
- 10.1089/hyb.2006.25.349
- PMID
- 17203997
- ISSN
- 1554-0014
- eISSN
- 1557-8348
- Language
- English
- Date published
- 12/2006
- Academic Unit
- Microbiology and Immunology; Pulmonary Medicine; Infectious Diseases; Stead Family Department of Pediatrics; Internal Medicine
- Record Identifier
- 9984093354702771
Metrics
14 Record Views