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United States Department of Agriculture

Agricultural Research Service

Title: The Production and Characterization of Anti-Bovine Cd14 Monoclonal Antibodies

Authors
item Sohn, Eun - UNIVERSITY OF MARYLAND
item Paape, Max
item Peters, Robert - UNIVERSITY OF MARYLAND
item Fetterer, Raymond
item Talbot, Neil
item Bannerman, Douglas

Submitted to: Veterinary Research
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: April 13, 2004
Publication Date: September 16, 2004
Citation: Sohn, E.J., Paape, M.J., Peters, R.P., Fetterer, R.H., Talbot, N.C., Bannerman, D.D. 2004. The production and characterization of anti-bovine cd14 monoclonal antibodies. Veterinary Research. 35(5):597-608.

Interpretive Summary: Researchers at the USDA-ARS research center in Beltsville, Maryland have developed a panel of monoclonal antibodies that are reactive with CD14. CD14 is found on the surface of a type of white blood cell called monocytes. When a cow becomes infected with coliform bacteria like Escherichia coli, toxins released by the bacteria bind to CD14 and causes the release of chemicals that make cows sick. About 40-50% of all clinical cases of mastitis, a disease of the mammary gland, are caused by coliform bacteria. Mastitis results in decreased milk production, increased veterinary costs, and culling and death of animals. Approximately 80% of all intramammary infections by coliform bacteria will result in clinical mastitis, and about 10% of the cows will develop acute clinical symptoms. Even in well-managed herds coliform mastitis continues to exist as an animal health problem. Antibiotic treatment, extensive fluid supplementation and metabolic support are not effective in relieving symptoms associated with clinical coliform mastitis. Anti-bovine CD14 mAb may provide a means of reducing clinical symptoms during acute coliform mastitis by blocking the binding of LPS to mCD14 , thus down-regulating release of inflammatory mediators.

Technical Abstract: To further characterize the chemical and biological properties of bovine soluble (bos) CD14, a panel of murine monoclonal antibodies (mAb) reactive with recombinant (r) bosCD14 were produced. A sandwich ELISA, using murine mAb and rabbit polyclonal antibodies each reactive with rbosCD14 was developed. All the mAb were reactive by ELISA with baculovirus-derived rbosCD14 and recognized rbosCD14 (43 kDa) by western blot analysis. The mAb also identified by western blot sCD14 (53 kDa) in milk and blood and sCD14 (48kDa) in a lysate of macrophages obtained from involuted bovine mammary gland secretions. Flow cytometric analysis showed that the mAb bound to macrophages isolated from involuted mammary gland secretions. 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 during mastitis.

Last Modified: 8/27/2014