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Title: THE PRODUCTION AND CHARACTERIZATION OF ANTI-BOVINE CD14 MONOCLONAE ANTIBODIES

Author
item SOHN, EUN - UNIVERSITY OF MARYLAND
item PETERS, ROBERT - UNIVERSITY OF MARYLAND
item Fetterer, Raymond
item Talbot, Neil
item Bannerman, Douglas

Submitted to: BARC Poster Day
Publication Type: Abstract Only
Publication Acceptance Date: 4/13/2004
Publication Date: 9/20/2004
Citation: Sohn, E.J., Peters, R., Fetterer, R.H., Talbot, N.C., Bannerman, D.D. 2004. The production and characterization of anti-bovine cd14 monoclonal antibodies. BARC Poster Day.

Interpretive Summary:

Technical Abstract: To characterize further the chemical and biological properties of bovine soluble (bos) CD14, a panel of ten murine monoclonal antibodies (mAb) reactive with recombinant (r) bosCD14 were produced. A sandwich ELISA, using murine mAb and rabbit polyclonal antibodies reactive with rbosCD14 was developed. All the mAb were reactive by ELISA with baculovirus-derived rbosCD14 and they recognized rbosCD14 (40 kDa) by western blot analysis. The mAb also identified by western blot sCD14 (53 kDa) in milk and blood and sCD14 (47kDa) in a lysate of macrophages obtained from involuted bovine mammary gland secretions. Analysis by ELISA of whey samples after intramammary injection of lipopolysaccharide (LPS) (100 µg) revealed increased sCD14 levels between 6 to 42 h after injection. Flow cytometric analysis showed that the mAb bound to macrophages isolated from involuted mammary gland secretions and mouse macrophages but not to swine or horse monocytes. Addition of anti-rbosCD14 mAb to monocytes stimulated with LPS reduced in vitro production of TNF-alpha. The anti-rbosCD14 antibodies generated in this study will be useful in studying CD14, an accessory molecule that contributes to host innate recognition of bacterial cell wall components in mammary secretions produced during mastitis.