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Assessment of epithelial cells' immune and inflammatory response to Staphylococcus aureus when exposed to a macrolide

Published online by Cambridge University Press:  08 September 2010

Maria Mazzilli  and
Alfonso Zecconi
Maria Mazzilli
Affiliation:
Department of Animal Pathology, Hygiene and Public Health, Università degli Studi di Milano, Via Celoria 10, 20133 Milano, Italy
Alfonso Zecconi*
Affiliation:
Department of Animal Pathology, Hygiene and Public Health, Università degli Studi di Milano, Via Celoria 10, 20133 Milano, Italy
*
*For correspondence; e-mail: [email protected]
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Abstract

Non-specific (innate) immune response plays a major role in defending the udder from bacterial invasion. Moreover, recent investigations suggest that mammary gland epithelial cells (MGEC) could have a large and important role as a source of soluble components of immune defences. Despite many attempts to find other ways to control/prevent mastitis (i.e. vaccine) antimicrobial therapy is still the most used and effective means of curing clinical and subclinical mastitis. However, drug concentrations and therapy durations are far from the optimal in order to reduce costs. Therefore, efficacy of antimicrobial therapy is dependent not only on the substance activity but also on the positive interactions with the host innate immune response. Surprisingly, information on these interactions is rather scarce in the mastitis field. A simple experimental model was developed based on BME-UV cell line, Staphylococcus aureus as a challenge and a macrolide as an antimicrobial to assess the interactions among epithelial cells, Staph. aureus and the potential effects of antimicrobials on the immune system. The results of this study confirmed that tylosin has good antimicrobial activity against both intracellular and extracellular Staph. aureus in bovine MGEC without affecting cell functions. In this study, a significant down-regulation of IL-1 and IL-6 was observed, while TNF and IL-8 expression rate numerically increased, but differences were not significant. To our knowledge, this is the first paper assessing the concentration of two lysosomal enzymes, lysozyme and N-acetyl-β-d-glucosaminidase (NAGase), in Staph. aureus-stimulated MGEC. The results of this study confirmed that tylosin could have a significant effect on the release of these enzymes. Moreover, even if both enzymes have a similar substrate as a target, the results suggest different secretion mechanisms and an influence of antimicrobial treatment on these mechanisms. Successful mastitis cure is the result of achieving the optimal efficiency of both innate immune defences and therapeutical activities, by means of killing bacteria without eliciting an excessive inflammatory response. Therefore, antimicrobials for mastitis therapy should be selected not only on bacterial sensitivity, but also for their positive interactions with the innate immune response of the mammary gland. This study showed that an in-vitro model based on Staph. aureus challenge on MGEC could be helpful in assessing both the intracellular and extracellular activity of antimicrobials and their influence on epithelial cell immune and inflammatory response.

Keywords

MastitismacrolidesStaphylococcus aureusimmunityinflammation
Type
Research Article
Information
Journal of Dairy Research , Volume 77 , Issue 4 , November 2010 , pp. 404 - 410
Copyright
Copyright © Proprietors of Journal of Dairy Research 2010

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References

Bazzi, C, Petrini, C, Rizza, V, Arrigo, G, Napodano, P, Paparella, M & D'Amico, G 2002 Urinary N-acetyl-beta-glucosaminidase excretion is a marker of tubular cell dysfunction and a predictor of outcome in primary glomerulonephritis. Nephrology Dialysis Transplantation 17 1890–1896CrossRefGoogle Scholar
Bolourchi, M, Hovareshti, P & Tabatabayi, AH 1995 Comparison of the effects of local and systemic dry cow therapy for staphylococcal mastitis control. Preventive Veterinary Medicine 25 63–67CrossRefGoogle Scholar
Bonnier, M, Dore, C, Amedeo, J & Guerin-Faublee, V 2006 In vitro activity of tylosin and tilmicosin against cocci isolated from bovine mastitis. Revue De Medecine Veterinaire 157 486–489Google Scholar
Burton, JL & Erskine, RJ 2003 Immunity and mastitis—some new ideas for an old disease. Veterinary Clinics of North America—Food Animal Practice 19 1–45CrossRefGoogle ScholarPubMed
Cao, XY, Dong, M, Shen, JZ, Wu, BB, Wu, CM, Du, XD, Wang, Z, Qi, YT & Li, BY 2006 Tilmicosin and tylosin have anti-inflammatory properties via modulation of COX-2 and iNOS gene expression and production of cytokines in LPS-induced macrophages and monocytes. International Journal of Antimicrobial Agents 27 431–438CrossRefGoogle ScholarPubMed
Chin, AC, Lee, WD, Murrin, KA, Morck, DW, Merrill, JK, Dick, P & Buret, AG 2000 Tilmicosin induces apoptosis in bovine peripheral neutrophils in the presence or in the absence of Pasteurella haemolytica and promotes neutrophil phagocytosis by macrophages. Antimicrobial Agents and Chemotherapy 44 2465–2470CrossRefGoogle ScholarPubMed
Cooper, GM & Hausman, RE 2007 The Cell: A Molecular Approach. Washington DC, USA: ASM PressGoogle ScholarPubMed
Dego, OK, van Dijk, JE & Nederbragt, H 2002 Factors involved in the early pathogenesis of bovine Staphylococcus aureus mastitis with emphasis on bacterial adhesion and invasion: a review. Veterinary Quarterly 24 181–198CrossRefGoogle Scholar
Didier, A & Kessel, S 2004 Novel in-vitro co-culture system for studies on leukocyte-mammary gland epithelial cell cross-talk. Milchwissenschaft-Milk Science International 59 236–239Google Scholar
Fitzgerald, DC, Meade, KG, McEvoy, AN, Lillis, L, Murphy, EP, Machugh, DE & Baird, AW 2007 Tumour necrosis factor-alpha (TNF-alpha) increases nuclear factor kappaB (NFkappaB) activity in and interleukin-8 (IL-8) release from bovine mammary epithelial cells. Veterinary Immunology and Immunopathology 116 59–68CrossRefGoogle ScholarPubMed
Ganz, T 2004 Antimicrobial polypeptides. Journal of Leukocyte Biology 75 34–38CrossRefGoogle ScholarPubMed
Giguere, S 2007 Macrolides, azalides and ketolides. In: Antimicrobial Therapy in Veterinary Medicine (Eds Giguere, S, Prescott, JF, Baggot, JD, Walker, RD & Dowling, PM) p. 626. Hoboken NJ, USA: Wiley-BlackwellGoogle Scholar
Hoeben, D, Burvenich, C & Heynemann, R 1997 Influence of antimicrobial agents on bactericidal activity of bovine polymorphonuclear leukocytes. Veterinary Immunology and Immunopathology 56 271–282CrossRefGoogle Scholar
Ianaro, A, Ialenti, A, Maffia, P, Sautebin, L, Rombola, L, Carnuccio, R, Iuvone, T, D'Acquisto, F & Di Rosa, M 2000 Anti-inflammatory activity of macrolide antibiotics. Journal of Pharmacology and Experimental Therapeutics 292 156–163Google ScholarPubMed
Kelly, BA & Carchman, RA 1987 The relationship between lysosomal enzyme release and protein phosphorylation in human monocytes stimulated by phorbol esters and opsonized zymosan. Journal of Biological Chemistry 262 17404–17411CrossRefGoogle ScholarPubMed
Kitchen, BJ, Middleton, G & Salmon, M 1978 Bovine milk N-acetyl-beta-d-glucosaminidase and its significance in the detection of abnormal udder secretions. Journal of Dairy Research 45 15–20CrossRefGoogle ScholarPubMed
Kitchen, BJ, Seng Kwee, W, Middleton, G & Andrews, RJ 1984 Relationship between the level of N-acetyl-β-d-glucodaminidase (NAGase) in bovine milk and the presence of mastitis pathogens. Journal of Dairy Research 51 11–16CrossRefGoogle ScholarPubMed
Labro, MT 2004a Cellular and molecular effects of macrolides on leukocyte function. Current Pharmaceutical Design 10 3067–3080CrossRefGoogle ScholarPubMed
Labro, MT 2004b Macrolide antibiotics: current and future uses. Expert Opinion on Pharmacotherapy 5 541–550CrossRefGoogle ScholarPubMed
Lammers, A, Nuijten, PJM, Kruijt, E, Stockhofe-Zurwieden, N, Vecht, U, Smith, HE & van Zijderveld, FG 1999 Cell tropism of Staphylococcus aureus in bovine mammary gland cell cultures. Veterinary Microbiology 67 77–89CrossRefGoogle ScholarPubMed
Leutenegger, CM, Alluwaimi, AM, Smith, WL, Perani, L & Cullor, JS 2000 Quantitation of bovine cytokine mRNA in milk cells of healthy cattle by real-time TaqMan® polymerase chain reaction. Veterinary Immunology and Immunopathology 77 275–287CrossRefGoogle ScholarPubMed
López-Boado, YS & Rubin, BK 2008 Macrolides as immunomodulatory medications for the therapy of chronic lung diseases. Current Opinion in Pharmacology 8 286–291CrossRefGoogle ScholarPubMed
NCCLS 2000 Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard. In: NCCLS document M7-A5 5th Edition, p. 36. Wayne PA, USA: NCCLSGoogle Scholar
NCCLS 2002 Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals; approved standard – second edition, NCCLS document M31-A2. In: NCCLS document M31-A2 2nd Edition, p. 86. Wayne PA, USA: NCCLSGoogle Scholar
Nickerson, SC, Paape, MJ & Dulin, AM 1985 Effect of antibiotics and vehicles on bovine polymorphonuclear leukocyte morphologic features, variability and phagocytic activity in vitro. American Journal of Veterinary Research 46 2259–2265Google Scholar
Pfaffl, MW, Horgan, GW & Dempfle, L 2002 Relative expression software tool (REST (c)) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Research 30 1–12CrossRefGoogle Scholar
Piccinini, R, Binda, E, Belotti, M, Daprà, V & Zecconi, A 2007 Evaluation of milk components during whole lactation in healthy quarters. Journal of Dairy Research 74 226–232CrossRefGoogle ScholarPubMed
Piccinini, R, Cesaris, L, Daprà, V, Borromeo, V, Picozzi, C, Secchi, C & Zecconi, A 2008 The role of teat skin contamination in the epidemiology of Staphylococcus aureus intramammary infections. Journal of Dairy Research 76 36–41CrossRefGoogle ScholarPubMed
Rainard, P & Riollet, C 2006 Innate immunity of the bovine mammary gland. Veterinary Research 37 369–400CrossRefGoogle ScholarPubMed
Scorneaux, B & Shryock, TR 1999 Intracellular accumulation, subcellular distribution and efflux of tilmicosin in bovine mammary, blood and lung ells. Journal of Dairy Science 82 1202–1212CrossRefGoogle Scholar
Wagner, S & Erskine, RJ 2006 Antimicrobial drug use in bovine mastitis. In: Antimicrobial Therapy in Veterinary Medicine (Eds Giguere, S, Prescott, JF & Desmond Baggot, J). Ames IA, USA: Iowa University PressGoogle Scholar
Zalewska-Kaszubska, J & Gorska, D 2001 Anti-inflammatory capabilities of macrolides. Pharmacological Research 44 451–454CrossRefGoogle ScholarPubMed
Zavizion, B, van Duffelen, M, Schaeffer, W & Politis, I 1996 Establishment and characterization of a bovine mammary epithelial cell line with unique properties. In Vitro Cell Development Biology Animal 32 138–148CrossRefGoogle ScholarPubMed
Zecconi, A 2007 Contagious mastitis control. FIL-IDF Bulletin 416 34–40Google Scholar
Zecconi, A, Binda, E, Borromeo, V & Piccinini, R 2005 Relationship between some Staphylococcus aureus pathogenic factors and growth rates or somatic cell counts. Journal of Dairy Research 72 203–208CrossRefGoogle ScholarPubMed
Zecconi, A, Cesaris, L, Liandris, E, Daprà, V & Piccinini, R 2006 Role of several Staphylococcus aureus virulence factors on the inflammatory response in bovine mammary gland. Microbial Pathogenesis 40 177–183CrossRefGoogle ScholarPubMed
Zecconi, A & Piccinini, R 2001 Efficacy of tylosin when administered during the drying-off period for the prevention and treatment of intramammary infections. In: 3rd Middle-European Congress for Buiatrics, pp. 97–100. Nove Mesto Check Rep 24–25/05/01: The Czech Association for BuiatricsGoogle Scholar
Zecconi, A & Piccinini, R 2002 Intramammary infections: epidemiology and diagnosis. In: XXII World Buiatric Congress— Recent developments and perspectives in bovine medicine, Hannover 18–23/08/2002 pp. 346–359. Eds Kaske, M, Scholz, H & Holtershinken, MGoogle Scholar
Zecconi, A, Piccinini, R, Bronzo, V & Casula, A 1996 Intracellular penetration of tylosin: influence on phagocytic activity and intracellular killing of Staph.aureus. In: World Buiatric Congress, Vol. 19, pp. 241–243. Edinburgh 8–12 July 1996Google Scholar
Zecconi, A, Piccinini, R & Guarini, CPB 1999 Tylosin in cows in the dry period. Obiettivi e Documenti Veterinari 20 49–54Google Scholar
Zecconi, A & Smith, KL 2003 Ruminant Mammary Gland Immunity. Bruxelles: FIL-IDFGoogle Scholar
Ziv, G 1980a Drug selection and use in mastitis: systemic vs local therapy. Journal of the American Veterinary Medical Association 176 1109–1115Google ScholarPubMed
Ziv, G 1980b Practical pharmacokinetic aspects of mastitis therapy–2: practical & therapeutic applications. Veterinary Medicine Small Animal Clinics 75 469–474Google ScholarPubMed