Czech J. Anim. Sci., 2012, 57(5):207-219 | DOI: 10.17221/5919-CJAS

Innate defense capability of challenged primary bovine mammary epithelial cells after an induced negative energy balance in vivo

K. Danowski1,2, D. Sorg1,2, J. Gross3, H.H.D. Meyer1,2, H. Kliem1,2
1 Physiology Weihenstephan, Technical University Munich, Freising-Weihenstephan, Germany
2 ZIEL - Research Centre for Nutrition and Food Sciences, Technical University Munich, Freising-Weihenstephan, Germany
3 Animal Nutrition, Technical University Munich, Freising-Weihenstephan, Germany

Negative energy balance (NEB), if followed by metabolic imbalance, is a common problem in high-yielding dairy cows frequently associated with inflammation of the mammary gland. After entering the teat canal, mammary epithelium is the first line of defense against a pathogen invasion. To investigate the effect of NEB on the innate host defense of the mammary epithelium, primary bovine mammary epithelial cell (pbMEC) cultures were generated by cell extraction of milk derived from energy restricted and control feeding cows. pbMEC were obtained from 8 high-yielding dairy cows affected by induced NEB in mid-lactation due to a reduction to 51 ± 2% of total energy requirement (restriction group) and from 7 control cows (control group). They were exposed to heat-inactivated Escherichia coli and Staphylococcus aureus for 24 and 72 h to investigate the influence of NEB on gene expression profiles of cytokines, chemokines, genes associated with apoptosis and antimicrobial peptides plus their receptors (AMPR) of the innate immune response. The immune challenge of pbMEC demonstrated an effect of immune capacity and NEB in 15 differential expressed genes. NEB induced a substantial up-regulation in restriction compared to control cells by trend in E. coli and a down-regulation in S. aureus exposed cells. Our investigations showed that the dietary-induced NEB in vivo influenced the immune response of pbMEC in vitro and altered the expression of immunological relevant genes due to a difference in energy supply. These results demonstrate that pbMEC are a suitable model for mastitis research, in which even effects of feeding regimes can be displayed.

Keywords: pbMEC; mastitis; energy deficit; E. coli; dairy cow; gene expression; innate immune response

Published: May 31, 2012  Show citation

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Danowski K, Sorg D, Gross J, Meyer HHD, Kliem H. Innate defense capability of challenged primary bovine mammary epithelial cells after an induced negative energy balance in vivo. Czech J. Anim. Sci. 2012;57(5):207-219. doi: 10.17221/5919-CJAS.
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References

  1. Akbar A.N., Borthwick N.J., Wickremasinghe G.R., Panayiotidis P., Pilling D., Bofill M., Krajewski S., Reed J.C., Salmon M. (1996): Interleukin-2 receptor common y-chain signaling cytokines regulate activated T cell apoptosis in response to growth factor withdrawal: Selective induction of anti-apoptotic (bcl-2, bcl-xL) but not pro-apoptotic (bax, bcl-xS) gene expression. European Journal of Immunology, 26, 294-299. Go to original source... Go to PubMed...
  2. Almeida P.F., Pokorny A. (2009): Mechanisms of antimicrobial, cytolytic, and cell-penetrating peptides: from kinetics to thermodynamics. Biochemistry, 48, 8083-8093. Go to original source... Go to PubMed...
  3. Bannerman D.D. (2009): Pathogen-dependent induction of cytokines and other soluble inflammatory mediators during intramammary infection of dairy cows. Journal of Animal Science, 87, 10-25. Go to original source... Go to PubMed...
  4. Bocchinfuso G., Palleschi A., Orioni B., Grande G., Formaggio F., Toniolo C., Park Y., Hahm K.S., Stella L. (2009): Different mechanisms of action of antimicrobial peptides: insights from fluorescence spectroscopy experiments and molecular dynamics simulations. Journal of Peptide Science, 15, 550-558. Go to original source... Go to PubMed...
  5. Bournazou I., Pound J.D., Duffin R., Bournazos S., Melville L.A., Brown S.B., Rossi A.G., Gregory C.D. (2009): Apoptotic human cells inhibit migration of granulocytes via release of lactoferrin. The Journal of Clinical Investigation, 119, 20-32. Go to original source... Go to PubMed...
  6. Boutinaud M., Jammes H. (2002): Potential uses of milk epithelial cells: a review. Reproduction Nutrition Development, 42, 133-147. Go to original source... Go to PubMed...
  7. Buitenhuis B., Rontved C.M., Edwards S.M., Ingvartsen K.L., Sorensen P. (2011): In depth analysis of genes and pathways of the mammary gland involved in the pathogenesis of bovine Escherichia coli-mastitis. BMC Genomics, 12, 130. Go to original source... Go to PubMed...
  8. Griesbeck-Zilch B., Meyer H.H., Kuhn C.H., Schwerin M., Wellnitz O. (2008): Staphylococcus aureus and Escherichia coli cause deviating expression profiles of cytokines and lactoferrin messenger ribonucleic acid in mammary epithelial cells. Journal of Dairy Science, 91, 2215-2224. Go to original source... Go to PubMed...
  9. Griesbeck-Zilch B., Osman M., Kuhn C., Schwerin M., Bruckmaier R.H., Pfaffl M.W., Hammerle-Fickinger A., Meyer H.H., Wellnitz O. (2009): Analysis of key molecules of the innate immune system in mammary epithelial cells isolated from marker-assisted and conventionally selected cattle. Journal of Dairy Science, 92, 4621-4633. Go to original source... Go to PubMed...
  10. Gross J., van Dorland H.A., Bruckmaier R.M., Schwarz F.J. (2011): Performance and metabolic profile of dairy cows during a lactational and deliberately induced negative energy balance with subsequent realimentation. Journal of Dairy Science, 94, 1820-1830. Go to original source... Go to PubMed...
  11. Gunther J., Koczan D., Yang W., Nurnberg G., Repsilber D., Schuberth H.J., Park Z., Maqbool N., Molenaar A., Seyfert H.M. (2009): Assessment of the immune capacity of mammary epithelial cells: comparison with mammary tissue after challenge with Escherichia coli. American Journal of Veterinary Research, 40, 31. Go to original source... Go to PubMed...
  12. Haston W.S., Shileds J.M. (1985): Neutrophil leucocyte chemotaxis: a simplified assay for measuring porlarizing responses to chemotactic factors. Journal of Immunological Methods, 81, 229-237. Go to original source... Go to PubMed...
  13. Kraus D., Peschel A. (2006): Molecular mechanisms of bacterial resistance to antimicrobial peptides. Current Topics in Microbiology and Immunology, 306, 231-250. Go to original source... Go to PubMed...
  14. Kubista M., Andrade J.M., Bengtsson M., Foroota A., Jonak J., Lind K., Sindelka R., Sjoback R., Sjogreen B., Strombom L., Stahlberg A., Zoric N. (2006): The realtime polymerase chain reaction. Molecular Aspects of Medicine, 27, 95-125. Go to original source... Go to PubMed...
  15. Lahouassa H., Moussay E., Rainard P., Riollet C. (2007): Differential cytokine and chemokine responses of bovine mammary epithelial cells to Staphylococcus aureus and Escherichia coli. Cytokine, 38, 12-21. Go to original source... Go to PubMed...
  16. Livak K.J., Schmittgen T.D. (2001): Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 25, 402-408. Go to original source... Go to PubMed...
  17. Loor J.J., Everts R.E., Bionaz M., Dann H.M., Morin D.E., Oliveira R., Rodriguez-Zas S.L., Drackley J.K., Lewin H.A. (2007): Nutrition-induced ketosis alters metabolic and signaling gene networks in liver of periparturient dairy cows. Physiological Genomics, 32, 105-116. Go to original source... Go to PubMed...
  18. Lutzow Y.C., Donaldson L., Gray C.P., Vuocolo T., Pearson R.D., Reverter A., Byrne K.A., Sheehy P.A., Windon R., Tellam R.L. (2008): Identification of immune genes and proteins involved in the response of bovine mammary tissue to Staphylococcus aureus infection. BMC Veterinary Research, 4, 18. Go to original source... Go to PubMed...
  19. Mehrzad J., Duchateau L., Burvenich C. (2005): Influence of resident milk neutrophils chemiluminescence and viability on the severity of bovine coliform mastitis. Veterinary Research, 36, 101-116. Go to original source... Go to PubMed...
  20. Molenaar A.J., Harris D.P., Rajan G.H., Pearson M.L., Callaghan M.R., Sommer L., Farr V.C., Oden K.E., Miles M.C., Petrova R.S., Good L.L., Singh K., McLaren R.D., Prosser C.G., Kim K.S., Wieliczko R.J., Dines M.H., Johannessen K.M., Grigor M.R., Davis S.R., Stelwagen K. (2009): The acute-phase protein serum amyloid A3 is expressed in the bovine mammary gland and plays a role in host defence. Biomarkers, 14, 26-37. Go to original source... Go to PubMed...
  21. Nunez G., Benedict M.A., Hu Y., Inohara N. (1998): Caspases: the proteases of the apoptotic pathway. Oncogene, 17, 3237-3245. Go to original source... Go to PubMed...
  22. Pareek R., Wellnitz O., Van Dorp R., Burton J., Kerr D. (2005): Immunorelevant gene expression in LPS-challenged bovine mammary epithelial cells. Journal of Applied Genetics, 46, 171-177. Go to PubMed...
  23. Petzl W., Zerbe H., Gunther J., Yang W., Seyfert H.M., Nurnberg G., Schuberth H.J. (2008): Escherichia coli, but not Staphylococcus aureus triggers an early increased expression of factors contributing to the innate immune defense in the udder of the cow. Veterinary Research, 39, 18. Go to original source... Go to PubMed...
  24. Riedmaier I., Becker C., Pfaffl M.W., Meyer H.H. (2009): The use of omic technologies for biomarker development to trace functions of anabolic agents. Journal of Chromatography A, 1216, 8192-8199. Go to original source... Go to PubMed...
  25. Roche J.R., Friggens N.C., Kay J.K., Fisher M.W., Stafford K.J., Berry D.P. (2009): Invited review: Body condition score and its association with dairy cow productivity, health, and welfare. Journal of Dairy Science, 92, 5769-5801. Go to original source... Go to PubMed...
  26. Roosen S., Exner K., Paul S., Schroder J.M., Kalm E., Looft C. (2004): Bovine beta-defensins: identification and characterization of novel bovine beta-defensin genes and their expression in mammary gland tissue. Mammalian Genome, 15, 834-842. Go to original source... Go to PubMed...
  27. Schroeder J.W. (2010): Bovine mastitis and milking management. Mastitis control programs, North Dakota State University and U.S. Department of Agriculture, NDSU Extension Service Circular AS-1129. Available from http://www.ag.ndsu.edu/pubs/ansci/dairy/as1129.pdf
  28. Sordillo L.M., Streicher K.L. (2002): Mammary gland immunity and mastitis susceptibility. Journal of Mammary Gland Biology and Neoplasia, 7, 135-146. Go to original source... Go to PubMed...
  29. Suriyasathaporn W., Heuer C., Noordhuizen-Stassen E.N., Schukken Y.H. (2000): Hyperketonemia and the impairment of udder defense: a review. Veterinary Research, 31, 397-412. Go to original source... Go to PubMed...
  30. Tesfaye G.Y., Regassa F.G., Kelay B. (2009): Milk yield and associated economic losses in quarters with subclinical mastitis due to Staphylococcus aureus in Ethiopian crossbred dairy cows. Tropical Animal Health and Production, 42, 925-931. Go to original source... Go to PubMed...
  31. Vorbach C., Capecchi M.R., Penninger J.M. (2006): Evolution of the mammary gland from the innate immune system? BioEssays, 28, 606-616. Go to original source... Go to PubMed...
  32. Wellnitz O., Kerr D.E. (2004): Cryopreserved bovine mammary cells to model epithelial response to infection. Veterinary Immunology and Immunopathology, 101, 191-202. Go to original source... Go to PubMed...
  33. Wellnitz O., Reith P., Haas S.C., Meyer H.H.D. (2006): Immune relevant gene expression of mammary epithelial cells and their influence on leukocytechemotaxis in response to different mastitis pathogens. Veterinarni Medicina, 51, 125-132. Go to original source...
  34. Werling D., Piercy J., Coffey T.J. (2006): Expression of toll-like receptors (TLR) by bovine antigen-presenting cells - potential role in pathogen discrimination? Veterinary Immunology and Immunopathology, 112, 2-11. Go to original source... Go to PubMed...
  35. Zhao X., Lacasse P. (2008): Mammary tissue damage during bovine mastitis: causes and control. Journal of Animal Science, 86, 57-65. Go to original source... Go to PubMed...
  36. Zasloff M. (2002): Antimicrobial peptides of multicellular organisms. Nature, 415, 389-395. Go to original source... Go to PubMed...

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