Skip to main content Accessibility help

Expression of acute phase proteins and inflammatory cytokines in mouse mammary gland following Staphylococcus aureus challenge and in response to milk accumulation

  • Sasan Nazemi (a1), Bent Aalbæk (a2), Mads Kjelgaard-Hansen (a1), Sina Safayi (a3), Dan Arne Klærke (a1) and Christopher H Knight (a1)...


We used a mouse model of pathogenic (Staphylococcus aureus) and non-pathogenic (teat sealing) mammary inflammation to investigate mRNA expression of several inflammatory cytokines and acute phase proteins (APP) in mammary tissue and liver, and the appearance of some of these factors in plasma and milk. The expression levels of IL1β and TNFα were markedly up-regulated in Staph. aureus-inoculated mammary tissue at 72 h, whilst IL6 was up-regulated to a lesser extent in a way which was not confined to the inoculated glands. APP expression was up-regulated at 48 and 72 h in both Staph. aureus-inoculated and teat-sealed mammary glands. These differences between cytokine and APP expression provide additional support for the contention that APPs are produced within the mammary tissue itself during inflammation, rather than in associated immune cells. We propose that measurement of cytokines and APP in combination might provide a tool for diagnostic discrimination between mastitis caused by pathogenic invasion and milk accumulation, and hence allow for better targeting of antibiotic therapy. In comparison with mammary expression, expression of cytokines in liver tissue was up-regulated to a similar or lesser extent, whilst expression of APP was up-regulated to a much greater extent. The first appearance of increased cytokine and APP concentrations in plasma and of milk amyloid A (MAA) in milk occurred in advance of the measurable up-regulation of expression, hence their origin cannot be stated with certainty.


Corresponding author

* For correspondence; e-mail:


Hide All
Akerstedt, M, Forsback, L, Larsen, L & Svennersten-Sjaunja, K 2011 Natural variation in biomarkers indicating mastitis in healthy cows. Journal of Dairy Research 78 8896
Anderson, JC 1977 Experimental staphylococcal mastitis in the mouse: the induction of chronic mastitis and its response to antibiotic therapy. Journal of Comparative Pathology 87 611621
Bannerman, DD 2009 Pathogen-dependent introduction of cytokines and other soluble inflammatory mediators during intramammary infection of dairy cows. Journal of Animal Science 87 1025
Bannerman, DD, Paape, MJ, Lee, J, Zhao, X, Hope, JC & Rainard, P 2004 Escherichia coli and staphylococcus aureus elicit differential innate immune response following intramammary infection. Clinical and Diagnostic Laboratory Immunology 11 463472
Bayles, KW, Wesson, CA, Liou, LE, Fox, LK, Bohanch, GA & Trumble, WR 1998 Intracellular Staphylococcus aureus escape the endosome and induces apoptosis in epithelial cells. Infection and Immunity 66 336342
Brouillette, E & Malouin, F 2005a The pathogenesis and control of Staphylococcus aureus-induced mastitis: study models in the mouse. Microbes and Infection 7 560568
Brouillette, E, Grondin, G, Lefebvre, C, Talbot, BG & Malouin, F 2004 Mouse mastitis model of infection for antimicrobial compound efficacy studies against intracellular and extracellular forms of Staphylococcus aureus . Veterinary Microbiology 101 253262
Brouillette, E, Grondin, G, Talbot, BG & Malouin, F 2005b Inflammatory cell infiltration as an indicator of Staphylococcus aureus infection and therapeutic efficacy in experimental mouse mastitis. Veterinary Immunology and Immupathology 104 163169
Ceciliani, F, Pocacqua, V, Provasi, E, Comunian, C, Bertolini, A, Bronzo, V, Moroni, P & Sartorelli, P 2005 Identification of the bovine α1-acid glycoprotein in colostrum and milk. Veterinary Research 36 735746
Ceciliani, F, Pocacqua, V, Lecchi, C, Fortin, R, Rebucci, R, Avallone, G, Bronzo, V, Cheli, F & Sartorelli, P 2007 Differential expression and secretion of a1-acid glycoprotein in bovine milk. Journal of Dairy Research 74 374380
Cha, E, Bar, D, Hertl, JA, Tauer, LW, Bennett, G, Gonzalez, RN, Schukken, YH, Welcome, FL & Grohn, YT 2011 The cost and management of different types of clinical mastitis in dairy cows by dynamic programming. Journal of Dairy Science 94 44764487
Chandler, RL 1970 Experimental bacterial mastitis in the mouse. Journal of Medical Microbiology 3 273289
Eckersall, PD 1995 Acute phase proteins as markers for inflammatory lesions. Comparative Haemotology International 5 9397
Eckersall, PD & Bell, R 2010 Acute phase proteins: biomarkers of infection and inflammation in veterinary medicine. Veterinary Journal 185 2327
Eckersall, PD, Young, FJ, Nolan, AM, Knight, CH, McComb, C, Waterson, MM, Hogarth, CJ, Scott, EM & Fitzpatrick, JL 2006 Acute phase proteins in bovine milk in an experimental model of Staphylococcus aureus subclinical mastitis. Journal of Dairy Science 89 14881501
Eckersall, PD, Lawson, FP, Bence, L, Waterson, MM, Lang, TL, Donachie, W & Fontains, MC 2007 Acute phase protein response in an experimental model of ovine caseous lymphadenitis. BMC Veterinary Research 3 35
Goldman, AS, Chheda, S, Garofalo, R & Schmalstieg, C 1996 Cytokines in human milk: properties and potential effects upon the mammary gland and the neonate. Journal of Mammary Gland Biology and Neoplasia 1 251258
Gunther, J, Katrin, E, Poschadel, N, Petzl, W, Holm, Z, Simone, M, Helmut, B & Hans-Martin, S 2011 Comparative kinetics of Escherichia coli- and Staphylococcus aureus-specific activation of key-immune pathways in mammary epithelial cell demonstrates that S. aureus elicits a delayed response dominated by Interleukin-6 (IL-6) but not by IL-1A or Tumor necrosis factor alpha. Infection and Immunity 79 695707
Han, LQ, Yang, GY, Zhu, HS, Wang, YY, Wang, LF, Guo, YJ, Lu, WF, Li, HJ & Wang, YL 2010 Selection and use of reference genes in mouse mammary glands. Genetics and Molecular Research 9 449456
Hensen, SM, Pavicic, MJ, Lohuis, JA, de Hoog, JA & Poutrel, B 2000 Location of Staphylococcus aureus within experimentally infected bovine udder and expression of capsular polysaccharide type 5 in situ. Journal of Dairy Science 83 19661975
Hiss, S, Mielenz, M, Bruckmaier, RM & Sauerwein, H 2004 Haptoglobin concentrations in blood and milk after endotoxin challenge and quantification of mammary Hp mRNA expression. Journal of Dairy Science 87 37783784
Hu, C, Gong, R, Guo, A & Chen, H 2010 Protective effect of ligand-binding domain of fibronectin-binding protein on mastitis induced by Staphylococcus aureus in mice. Vaccine 28 40384044
Koj, A 1996 Initiation of acute phase response and synthesis of cytokines. Biochimica et Biophysica Acta 1317 8494
Kovacevic-Filipovic, M, Ilic, V, Vujcic, Z, Dojnov, B, Stevanov-Pavlovic, M, Mijacevic, Z & Bozic, T 2012 Serum amyloid A isoforms in serum and milk from cows with Staphylococcus aureus subclinical mastitis. Veterinary Immunology and Immunopathology 145 120128
Kvist, LJ 2013 Re-examination of old truths: replication of a study to measure the incidence of lactational mastitis in breastfeeding women. International Breastfeeding Journal 8 2
Lakic, B, Sjaunja, KS, Norell, L, Dernfalk, J & Ostensson, K 2011 The effect of a single prolonged milking interval on inflammatory parameters, milk composition and yield in dairy cows. Veterinary Immunology and Immunopathology 140 110118
Lai, IH, Tsao, JH, Lu, YP, Lee, JW, Zhao, X, Chien, FL & Mao, SJ 2009 Neutrophils as one of the major haptoglobin sources in mastitis affected milk. Veterinary Research 40 17
McDonald, TL, Larson, MA, Mack, DR & Weber, A 2001 Elevated extrahepatic expression and secretion of mammary-associated serum amyloid A 3 (M-SAA3) into colostrum. Veterinary Immunology and Immunopathology 83 203211
Murata, H, Shimada, N & Yoshioka, M 2004 Current research on acute phase proteins in veterinary diagnosis: an overview. Veterinary Journal 168 2840
Nielsen, BH, Jacobsen, S, Andersen, PH, Niewold, TA & Heegaard, PMH 2004 Acute phase protein concentration in serum and milk from healthy cows, cows with clinical mastitis and cows with extramammary inflammatory conditions. Veterinary Record 154 361365
Pedersen, LH, Aalbæk, B, Røntved, CM, Ingvartsen, KL, Sorensen, NS, Heegaard, PM & Jensen, HE 2003 Early pathogenesis of inflammatory response in experimental bovine mastitis due to Streptococcus uberis . Journal of Comparative Pathology 128 156164
Peli, A, Scagliarini, A, Britti, D & Boari, A 2003 Detection of proinflammatory and regulatory cytokines in bovine milk using RT-PCR. Veterinary Research Communication 27 779781
Petzl, W, Zerbe, H, Gunther, J, Yang, W, Seyfert, H, Nurnberg, G & Schuberth, H 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
Pfaffl, MW 2001 A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Research 29 e45
Quarrie, LH, Addey, CV & Wilde, CJ 1996 Programmed cell death during mammary tissue involution induced by weaning, litter removal, and milk stasis. Journal of Cellular Physiology 168 559569
Rahbek, M, Nazemi, S, Ødum, L, Gupta, S, Poulsen, SS, Hay-Schidt, A & Klærke, DA 2014 Expression of the small conductance Ca2+-activated potassium channel subtype 3 (SK3) in rat uterus after stimulation with 17-β-estradiol. PLoS ONE 9 e87652
Rahman, MM, Miranda-Ribera, A, Lecchi, C, Bronzo, V, Sartorelli, P, Franciosi, F & Ceciliani, F 2008 Alpha1-acid glycoprotein is contained in bovine neutrophil granules and released after activation. Veterinary Immunology and Immunopathology 125 7181
Roberson, JR 2012 Treatment of clinical mastitis. Veterinary Clinics of North America: Food Animal Practice 28 271288
Schukken, YH, Gunther, J, Fitzpatrick, J, Fontaine, MC, Goetze, L, Holst, O, Leigh, J, Petzl, W, Schuberh, HJ, Sipka, A, Smith, DGE, Quesnell, R, Watts, J, Yancey, R, Zerbe, H, Gurjar, A, Zadoks, RN & Seyfert, HM; members of the Pfizer mastitis research consortium 2011 Host-response patterns of intramammary infection in dairy cows. Veterinary Immunology and Immunopathology 144 270289
Scott, JA, Robertson, M, Fitzpatrick, J, Knight, C & Mulholland, S 2008 Occurrence of lactational mastitis and medical management: a prospective cohort study in Glasgow. International Breastfeeding Journal 3 21
Seegers, H, Fourichon, C & Beaudeau, F 2003 Production effects related to mastitis and mastitis economics in dairy cattle herds. Veterinary Research 34 475491
Shipman, LJ, Docherty, AH, Knight, CH & Wilde, CJ 1987 Metabolic adaptations in mouse mammary gland during a normal lactation cycle and in extended lactation. Quarterly Journal of Experimental Physiology 72 303311
Stein, T, Morris, JS, Davies, CR, Weber-Hall, SJ, Duffy, M, Heath, VJ, Bell, AK, Ferrier, RK, Sandilands, GP & Gusterson, BA 2004 Involution of the mouse mammary gland is associated with an immune cascade and an acute-phase response, involving LBP, CD14 and STAT3. Breast Cancer Research 6 7591
Theil, PK, Labouriau, R, Sejrsen, K, Thomsen, B & Sørensen, MT 2005 Expression of gene involved in regulation of cell turnover during milk stasis and lactation rescue in sow mammary glands. Journal of Animal Science 83 23492356
Villalba, M, Hott, M, Martin, C, Aguila, B, Valdiva, S, Quezada, C, Zambrano, A, Concha, MI & Otth, C 2012 Herpes simplex virus type 1 induces simultaneous activation of toll-like receptors 2 and 4 and expression of the endogenous ligand serum amyloid A in astrocytes. Medical Microbiology and Immunology 201 371379
Wang, S, Diao, H, Guan, Q, Cruikshank, WW, Delovitch, TL, Jevnikar, AM & Du, C 2008 Decreased renal ischemia-reperfusion injury by IL-16 inactivation. Kidney International 73 318326
Yoshioka, M, Watanabe, A, Shimada, N, Murata, H, Yokomizo, Y & Nakajima, Y 2002 Regulation of haptoglobin secretion by recombinant bovine cytokines in primary cultured bovine hepatocytes. Domestic Animal Endocrinology 23 425433


Expression of acute phase proteins and inflammatory cytokines in mouse mammary gland following Staphylococcus aureus challenge and in response to milk accumulation

  • Sasan Nazemi (a1), Bent Aalbæk (a2), Mads Kjelgaard-Hansen (a1), Sina Safayi (a3), Dan Arne Klærke (a1) and Christopher H Knight (a1)...


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed