Hostname: page-component-848d4c4894-hfldf Total loading time: 0 Render date: 2024-06-01T09:02:29.766Z Has data issue: false hasContentIssue false
  • English
  • Français

Cell population, viability, and some key immunomodulatory molecules in different milk somatic cell samples in dairy cows

Published online by Cambridge University Press:  18 May 2009

Amandine Baumert ,
Rupert M. Bruckmaier  and
Olga Wellnitz
Amandine Baumert
Affiliation:
Veterinary Physiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland
Rupert M. Bruckmaier
Affiliation:
Veterinary Physiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland
Olga Wellnitz*
Affiliation:
Veterinary Physiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland
*
*For correspondence; e-mail: olga.wellnitz@physio.unibe.ch
Get access Rights & Permissions [Opens in a new window]

Abstract

Immune cells in the milk are most important in combating pathogens that invade the mammary gland. This study investigated the immune competence and viability of somatic milk cells that are already resident in milk and udders free of infection. Cells were studied in freshly removed milk to simulate conditions in the udder. Effects of incubation, cell preparation, and immunological stimulation with 0·5 μg/ml lipopolysaccharide (LPS) from Escherichia coli were analysed. Viability and differential counts of milk cells between high and low somatic cell count (SCC) quarters, and cisternal and alveolar milk with and without LPS stimulation were compared. Incubation and preparation of cells caused a cell loss which further increased with time independently of SCC and milk fraction. The viability of these cells was stable until 3 h post incubation and decreased until 6 h. Cell populations differed between both investigations, but did not change during the course of the experiment. mRNA expression of immune and apoptosis factors of the cells, measured by qPCR, did not change substantially: mRNA expression of caspase 3, Toll like receptor 4, and GM-CSF did not change, whereas the expression of the death receptor Fas/APO-1 (CD95), lactoferrin and lysozyme was decreased at 6 h. Cyclooxygenase-2 and TNF-α mRNA expression were decreased after 6 h of LPS treatment. In comparison with other studies in vivo or in vitro (in cell culture), in this study where cells are studied ex vivo (removed from the udder but kept in their natural environment, the milk) resident milk cells seem to be more vulnerable, less viable, less able to respond to stimulation, and thus less immune competent compared with cells that have freshly migrated from blood into milk after pathogen stimulation. The cell viability and differential cell count differed between high- and low-SCC milk and between cisternal and alveolar milk depending on the individual cow. In conclusion, the results support the view that for a most effective defence against invading pathogens the mammary gland is reliant on the recruitment of fresh immune cells from the blood.

Keywords

Immunitymammarymilk fractions
Type
Research Article
Information
Journal of Dairy Research , Volume 76 , Issue 3 , August 2009 , pp. 356 - 364
Copyright
Copyright © Proprietors of Journal of Dairy Research 2009

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Bannerman, DD, Paape, MJ, Lee, JW, Zhao, X, Hope, JP & Rainard, C 2004 Escherichia coli and Staphylococcus aureus elicit differential innate immune responses following intramammary infection. Clinical and Diagnostic Laboratory Immunology 11 463–472Google ScholarPubMed
Boudjellab, N, Chan-Tang, HS, Li, HS & Zhao, X 1998 Interleukin 8 response by bovine mammary epithelial cells to lipopolysaccharide stimulation. American Journal of Veterinary Research 59 1563–1567CrossRefGoogle ScholarPubMed
Brolund, L 1985 Cell counts in bovine milk. Causes of variation and applicability for diagnosis of subclinical mastitis. Acta Veterinaria Scandinavica Supplementum 80 1–123Google ScholarPubMed
Bruckmaier, RM & Blum, JW 1996 Simultaneous recording of oxytocin release, milk ejection and milk flow during milking of dairy cows with and without prestimulation. Journal of Dairy Research 63 201–208CrossRefGoogle ScholarPubMed
Burvenich, C, Paape, MJ, Hill, AW, Guidry, AJ, Miller, RH, Heyneman, R, Kremer, WD & Brand, A 1994 Role of the neutrophil leucocyte in the local and systemic reactions during experimentally induced E. coli mastitis in cows immediately after calving. Veterinary Quarterly 16 45–50CrossRefGoogle ScholarPubMed
Burvenich, C, Bannerman, DD, Lippolis, JD, Peelman, L, Nonnecke, BJ, Kehrli, ME Jr & Paape, MJ 2007 Cumulative physiological events influence the inflammatory response of the bovine udder to Escherichia coli infections during the transition period. Journal of Dairy Science 90 Suppl 1 39–54CrossRefGoogle ScholarPubMed
Carlson, GP & Kaneko, JJ 1975 Intravascular granulocyte kinetics in developing calves. American Journal of Veterinary Research 36 421–425Google ScholarPubMed
Colotta, F, Re, F, Polentarutti, N, Sozzani, S & Mantovani, A 1992 Modulation of granulocyte survival and programmed cell death by cytokines and bacterial products. Blood 80 2012–2020CrossRefGoogle ScholarPubMed
Didier, A & Bruckmaier, RM 2004 mRNA expression of apoptosis-related genes in mammary tissue and milk cells in response to lipopolysaccharide challenge and during subclinical mastitis. Milchwissenschaft 59 119–123Google Scholar
Duvall, E & Willie, AH 1986 Death and the cell. Immunology Today 7 115–119CrossRefGoogle ScholarPubMed
Fadeel, B, Ahlin, A, Henter, JI, Orrenius, S & Hampton, MB 1998 Involvement of caspases in neutrophil apoptosis: regulation by reactive oxygen species. Blood 92 4808–4818CrossRefGoogle ScholarPubMed
Hachiya, O, Takeda, Y, Miyata, H, Watanabe, H, Yamashita, T & Sendo, F 1995 Inhibition by bacterial lipopolysaccharide of spontaneous and TNF-alpha-induced human neutrophil apoptosis in vitro. Microbiology and Immunology 39 715–723CrossRefGoogle ScholarPubMed
Harmon, RJ & Heald, CW 1982 Migration of polymorphonuclear leukocytes into the bovine mammary gland during experimentally induced Staphylococcus aureus mastitis. American Journal of Veterinary Research 43 992–998Google ScholarPubMed
Harmon, RJ 1994 Physiology of mastitis and factors affecting somatic cell counts. Journal of Dairy Science 77 2103–2112CrossRefGoogle ScholarPubMed
Jacobsen, S, Toelboell, T & Andersen, PH 2005 Dose dependency and individual variability in selected clinical, haematological and blood biochemical responses after systemic lipopolysaccharide challenge in cattle. Veterinary Research 36 167–178CrossRefGoogle ScholarPubMed
Kehrli, EM & Shuster, DE 1994 Factors affecting milk somatic cells and their role in health of the bovine mammary gland. Journal of Dairy Science 77 619–627CrossRefGoogle ScholarPubMed
Lee, JW, Bannerman, DD, Paape, MJ, Huang, MK & Zhao, X 2006 Characterization of cytokine expression in milk somatic cells during intramammary infections with Escherichia coli or Staphylococcus aureus by real-time PCR. Veterinary Research 37 219–229CrossRefGoogle ScholarPubMed
Mangan, DF, Welch, GR & Wahl, SM 1991 Lipopolysaccharide, tumor necrosis factor-alpha, and IL-1 beta prevent programmed cell death (apoptosis) in human peripheral blood monocytes. Journal of Immunology 146 1541–1546CrossRefGoogle ScholarPubMed
Mehrzad, J, Dosogne, H, Meyer, E & Burvenich, C 2001a Local and systemic effects of endotoxin mastitis on the chemiluminescence of milk and blood neutrophils in dairy cows. Veterinary Research 32 131–144CrossRefGoogle ScholarPubMed
Mehrzad, J, Duchateau, L & Burvenich, C 2004 Viability of milk neutrophils and severity of bovine coliform mastitis. Journal of Dairy Science 87 4150–4162CrossRefGoogle ScholarPubMed
Mehrzad, J, Dosogne, H, Meyer, E, Heyneman, R & Burvenich, C 2001b Respiratory burst activity of blood and milk neutrophils in dairy cows during different stages of lactation. Journal of Dairy Research 68 399–415CrossRefGoogle ScholarPubMed
Mehrzad, J, Duchateau, L, Pyörälä, S & Burvenich, C 2002 Blood and milk neutrophil chemiluminescence and viability in primiparous and pluriparous dairy cows during late pregnancy, around parturition and early lactation. Journal of Dairy Science 85 3268–3276CrossRefGoogle ScholarPubMed
Menaker, RJ & Jones, NL 2003 Fascination with bacteria-triggered cell death: the significance of Fas-mediated apoptosis during bacterial infection in vivo. Microbes and Infection 5 1149–1158CrossRefGoogle ScholarPubMed
Newbould, FH 1973 The effect of added serum and glucose, and some inherent factors, on phagocytosis in vitro by milk leukocytes from several cows. Canadian Journal of Comparative Medicine 37 189–194Google ScholarPubMed
Olde Riekerink, RG, Barkema, HW, Veenstra, W, Berg, FE, Stryhn, H & Zadoks, RN 2007 Somatic cell count during and between milkings. Journal of Dairy Science 90 3733–3741CrossRefGoogle ScholarPubMed
Paape, MJ, Guidry, AJ, Kirk, ST & Bolt, DJ 1975 Measurement of phagocytosis of 32P-labeled Staphylococcus aureus by bovine leukocytes: lysostaphin digestion and inhibitory effect of cream. American Journal of Veterinary Research 36 1737–1743Google ScholarPubMed
Paape, MJ & Guidry, AJ 1977 Effect of fat and casein on intracellular killing of Staphylococcus aureus by milk leukocytes. Proceeding of the Society for Experimental Biology and Medicine 155 588–593CrossRefGoogle ScholarPubMed
Paape, M, Mehrzad, J, Zhao, X, Detilleux, J & Burvenich, C 2002 Defence of the bovine mammary gland by polymorphonuclear neutrophil leukocytes. Journal of Mammary Gland Biology and Neoplasia 7 109–121CrossRefGoogle ScholarPubMed
Paape, MJ, Bannerman, DD, Zhao, X & Lee, JW 2003 The bovine neutrophil: Structure and function in blood and milk. Veterinary Research 34 597–627Google ScholarPubMed
Persson-Waller, K 2000 Mammary gland immunology around parturition. Influence of stress, nutrition and genetics. Advances in Experimental Medicine and Biology 480 231–245CrossRefGoogle Scholar
Pfaffl, MW, Wittmann, SL, Meyer, HH & Bruckmaier, RM 2003 Gene expression of immunologically important factors in blood cells, milk cells and mammary tissue of cows. Journal of Dairy Science 86 538–545CrossRefGoogle ScholarPubMed
Prgomet, C, Sarikaya, H, Bruckmaier, RM & Pfaffl, MW 2005 Short-term effects on pro- inflammatory cytokine, Lactoferrin and CD14 mRNA expression levels in bovine immunoseparated milk and blood cells treated by LPS. Journal of Veterinary Medicine A, Physiology, Pathology, Clinical Medicine A52 317–324CrossRefGoogle Scholar
Rainard, P & Riollet, C 2006 Innate immunity of the bovine mammary gland. Veterinary Research 37 369–400CrossRefGoogle ScholarPubMed
Rysánek, D, Sládek, Z, Vasícková, D & Faldyna, M 2005 Effects of certain inducers of leukocytes migration into the bovine mammary gland on neutrophil apoptosis manifestation in a subsequent in vitro cultivation. Physiological Research 54 305–312CrossRefGoogle Scholar
Robertson, JD, Orrenius, S & Zhivotovsky, B 2000 Review: nuclear events in apoptosis. Journal of Structural Biology 129 246–358CrossRefGoogle ScholarPubMed
Sarikaya, H, Prgomet, C, Pfaffl, MW & Bruckmaier, RM 2004 Differentiation of leukocytes in bovine milk. Milchwissenschaft 59 586–589Google Scholar
Sarikaya, H, Werner-Misof, C, Atzkern, M & Bruckmaier, RM 2005 Distribution of leucocyte populations, and milk composition, in milk fractions of healthy quarters in dairy cows. Journal of Dairy Research 72 486–492CrossRefGoogle ScholarPubMed
Sarikaya, H, Schlamberger, G, Meyer, HH & Bruckmaier, RM 2006 Leukocyte populations and mRNA expression of inflammatory factors in quarter milk fractions at different somatic cell score levels in dairy cows. Journal of Dairy Science 89 2479–2486CrossRefGoogle ScholarPubMed
Schmitz, S, Pfaffl, MW, Meyer, HH & Bruckmaier, RM 2004 Short-term changes of mRNA expression of various inflammatory factors and milk proteins in mammary tissue during LPS-induced mastitis. Domestic Animal Endocrinology 26 111–126CrossRefGoogle ScholarPubMed
Schröder, AC & Hamann, J 2005 The influence of technical factors on differential cell count in milk. Journal of Dairy Research 72 153–158CrossRefGoogle ScholarPubMed
Sládek, Z & Ryšánek, D 1999 Ultrastructure of phagocytes from mammary glands of non-pregnant heifers. Anatomia Histologia Embryologia 28 291–297CrossRefGoogle ScholarPubMed
Sládek, Z & Ryšánek, D 2000 Apoptosis of polymorphonuclear leukocytes of the juvenile bovine mammary gland during induced influx. Veterinary Research 31 553–563CrossRefGoogle ScholarPubMed
Smits, E, Burvenich, C, Guidry, AJ, Heyneman, R & Massart-Leen, A 1999 Diapedesis across mammary epithelium reduces phagocytic and oxidative burst of bovine neutrophils. Veterinary Immunology and Immunopathology 68 169–176CrossRefGoogle ScholarPubMed
Sordillo, LM & Streicher, KL 2002 Mammary gland immunity and mastitis susceptibility. Journal of Mammary Gland Biology and Neoplasia 7 135–146CrossRefGoogle ScholarPubMed
Suriyasathaporn, W, Schukken, YH, Nielen, M & Brand, A 2000 Low somatic cell count: a risk factor for subsequent clinical mastitis in a dairy herd. Journal of Dairy Science 83 1248–1255CrossRefGoogle Scholar
Takeuchi, O, Hoshino, K, Kawai, T, Sanjo, H, Takada, H, Ogawa, T, Takeda, K & Akira, S 1999 Differential roles of TLR2 and TLR4 in recognition of gram-negative and gram-positive bacterial cell wall components. Immunity 11 443–451CrossRefGoogle ScholarPubMed
Van Oostveldt, K, Tomita, GM, Paape, MJ, Capuco, AV & Burvenich, C 2002a Apoptosis of bovine neutrophils during mastitis experimentally induced with Escherichia coli or endotoxin. American Journal of Veterinary Research 63 448–453CrossRefGoogle ScholarPubMed
Van Oostveldt, K, Paape, MJ & Burvenich, C 2002b Apoptosis of bovine neutrophils following diapedesis through a monolayer of endothelial and mammary epithelial cells. Journal of Dairy Science 85 139–147CrossRefGoogle ScholarPubMed
Wellnitz, O & Kerr, DE 2004 Cryopreserved bovine mammary cells to mode epithelial response to infection. Veterinary Immunology and Immunopathology 101 191–202CrossRefGoogle Scholar
Werling, D, Piercy, J & Coffey, TJ 2006 Expression of TOLL-like receptor (TLR) by bovine antigen-presenting cells — Potential role in pathogen discrimination? Veterinary Immunology and Immunopathology 112 2–11CrossRefGoogle ScholarPubMed