Mastitis Increases Mammary mRNA Abundance of β-Defensin 5, Toll-Like-Receptor 2 (TLR2), and TLR4 but Not TLR9 in Cattle

American Society for Microbiology - Tập 11 Số 1 - Trang 174-185 - 2004
Tom Goldammer1,2,3,4,5, Holm Zerbe1,2,3,4,6, Adrian Molenaar1,2,3,4,6, H. J. Schuberth1,2,3,4,6, Ronald M. Brunner1,2,3,4,6, S.R. Kata1,2,3,4,6, Hans‐Martin Seyfert1,2,3,4,6
1Clinic for Bovine Obstetrics and Gynecology
2Dairy Biotechnology Group, AgResearch, Ruakura Research Centre, Hamilton, New Zealand
3Department of Veterinary Pathobiology, Texas A&M University, College Station, Texas 77843
4Immunology Unit, School of Veterinary Medicine, 30173 Hannover, Germany
5Molecular Biology Research Division, Research Institute for the Biology of Farm Animals, 18196 Dummerstorf, Germany
6Molecular Biology Research Division, Research Institute for the Biology of Farm Animals, 18196 Dummerstorf

Tóm tắt

ABSTRACTCoordination of the primary defense mechanisms against pathogens relies on the appropriate expression of pathogen recognition receptors (PRRs) triggering the early release of effector molecules of the innate immune system. To analyze the impact of this system on the counteraction of infections of the mammary gland (mastitis), we characterized the bovine gene encoding the key PRR Toll-like receptor 9 (TLR9) and mapped its precise position on chromosome BTA22. The sequence information was used to establish real-time PCR quantification assays to measure the mRNA abundances of TLR9, TLR2, and TLR4 together with those of β-defensin 5 (BNBD5), an early bactericidal effector molecule of the innate system, in healthy and infected mammary glands. Mastitis strongly increased (4- to 13-fold) the mRNA abundances of all of these genes except TLR9. Slight subclinical infections already caused a substantial increase in the copy numbers, though they did so the least for TLR9. Induction was not systemic, since mRNA abundance was low in uninfected control quarters of the udder but high in the severely infected quarters of the same animal. The number of TLR2 copies correlated well with those of TLR4, indicating coordinated regulation of these two PRRs during infection of the udder. Their coordinated regulation explains our unexpected observation that pureStaphylococcus aureusinfections caused a strong increase also in TLR4 mRNA abundance. In situ hybridizations revealed that BNBD5 is expressed predominantly in the mammary epithelial cells (MEC) of the infected gland. Our data therefore suggest a significant contribution of the innate immune system to counteract mastitis and attribute a prominent effector function to the MEC.

Từ khóa


Tài liệu tham khảo

Aderem, A., and R. J. Ulevitch. 2000. Toll-like receptors in the induction of the innate immune response. Nature406:782-787.

Ashwell, M. S., C. E. Rexroad, Jr., R. H. Miller, and P. M. VanRaden. 1996. Mapping economic trait loci for somatic cell score in Holstein cattle using microsatellite markers and selective genotyping. Anim. Genet.27:235-242.

Band, M. R., J. H. Larson, M. Rebeiz, C. A. Green, D. W. Heyen, J. Donovan, R. Windish, C. Steining, P. Mahyuddin, J. E. Womack, and H. A. Lewin. 2000. An ordered comparative map of the cattle and human genomes. Genome Res.10:1359-1368.

Bauer, S., C. J. Kirschning, H. Hacker, V. Redecke, S. Hausmann, S. Akira, H. Wagner, and G. B. Lipford. 2001. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. Proc. Natl. Acad. Sci. USA98:9237-9242.

Bendelac, A., and D. T. Fearon. 2000. Innate immunity: receptors and effectors of innate immunity. Curr. Opin. Immunol.12:11-12.

Birchler, T., R. Seibl, K. Buchner, S. Loeliger, R. Seger, J. P. Hossle, A. Aguzzi, and R. P. Lauener. 2001. Human Toll-like receptor 2 mediates induction of the antimicrobial peptide human beta-defensin 2 in response to bacterial lipoprotein. Eur. J. Immunol.31:3131-3137.

Campos, M. A., I. C. Almeida, O. Takeuchi, S. Akira, E. P. Valente, D. O. Procopio, L. R. Travassos, J. A. Smith, D. T. Golenbock, and R. T. Gazzinelli. 2001. Activation of Toll-like receptor-2 by glycosylphosphatidylinositol anchors from a protozoan parasite. J. Immunol.167:416-423.

Carl, V. S., K. Brown-Steinke, M. J. Nicklin, and M. F. Smith, Jr. 2002. Toll-like receptor 2 and 4 (TLR2 and TLR4) agonists differentially regulate secretory interleukin-1 receptor antagonist gene expression in macrophages. J. Biol. Chem.277:17448-17456.

Chevalet, C., and F. Corpet. 1986. Statistical decision rules concerning synteny or independence between markers. Cytogenet. Cell Genet.43:132-139.

Chuang, T. H., and R. J. Ulevitch. 2000. Cloning and characterization of a sub-family of human Toll-like receptors: hTLR7, hTLR8 and hTLR9. Eur. Cytokine Netw.11:372-378.

Cribiu, E. P., D. Di Berardino, G. P. Di Meo, A. Eggen, D. S. Gallagher, I. Gustavsson, H. Hayes, L. Iannuzzi, C. P. Popescu, J. Rubes, S. Schmutz, G. Stranzinger, A. Vaiman, and J. Womack. 2001. International System for Chromosome Nomenclature of Domestic Bovids (ISCNDB 2000). Cytogenet. Cell Genet.92:283-299.

Dabbagh, K., and D. B. Lewis. 2003. Toll-like receptors and T-helper-1/T-helper-2 responses. Curr. Opin. Infect. Dis.16:199-204.

10.1128/IAI.68.1.113-119.2000

Diamond, G., M. Zasloff, H. Eck, M. Brasseur, W. L. Maloy, and C. L. Bevins. 1991. Tracheal antimicrobial peptide, a cysteine-rich peptide from mammalian tracheal mucosa: peptide isolation and cloning of a cDNA. Proc. Natl. Acad. Sci. USA88:3952-3956.

Dosogne, H., F. Vangroenweghe, and C. Burvenich. 2002. Potential mechanism of action of J5 vaccine in protection against severe bovine coliform mastitis. Vet. Res.33:1-12.

Du, X., A. Poltorak, Y. Wei, and B. Beutler. 2000. Three novel mammalian toll-like receptors: gene structure, expression, and evolution. Eur. Cytokine Netw.11:362-371.

Erskine, R. J., R. J. Eberhart, L. J. Hutchinson, S. B. Spencer, and M. A. Campbell. 1988. Incidence and types of clinical mastitis in dairy herds with high and low somatic cell counts. J. Am. Vet. Med. Assoc.192:761-765.

Goldammer, T., S. R. Kata, R. M. Brunner, U. Dorroch, H. Sanftleben, M. Schwerin, and J. E. Womack. 2002. A comparative radiation hybrid map of bovine chromosome 18 and homologous chromosomes in human and mice. Proc. Natl. Acad. Sci. USA99:2106-2111.

Hayashi, F., K. D. Smith, A. Ozinsky, T. R. Hawn, E. C. Yi, D. R. Goodlett, J. K. Eng, S. Akira, D. M. Underhill, and A. Aderem. 2001. The innate immune response to bacterial flagellin is mediated by Toll-like receptor 5. Nature410:1099-1103.

Hemmi, H., O. Takeuchi, T. Kawai, T. Kaisho, S. Sato, H. Sanjo, M. Matsumoto, K. Hoshino, H. Wagner, K. Takeda, and S. Akira. 2000. A Toll-like receptor recognizes bacterial DNA. Nature408:740-745.

Hibbitt, K. G., J. Brownlie, and C. B. Cole. 1971. The antimicrobial activity of cationic proteins isolated from the cells in bulk milk samples. J. Hyg. (London)69:61-68.

Hill, A. W. 1981. Factors influencing the outcome of Escherichia coli mastitis in the dairy cow. Res. Vet. Sci.31:107-112.

Hill, A. W., A. L. Shears, and K. G. Hibbitt. 1979. The pathogenesis of experimental Escherichia coli mastitis in newly calved dairy cows. Res. Vet. Sci.26:97-101.

Hirschfeld, M., Y. Ma, J. H. Weis, S. N. Vogel, and J. J. Weis. 2000. Cutting edge: repurification of lipopolysaccharide eliminates signaling through both human and murine toll-like receptor 2. J. Immunol.165:618-622.

Hoffmann, J. A., F. C. Kafatos, C. A. Janeway, and R. A. Ezekowitz. 1999. Phylogenetic perspectives in innate immunity. Science284:1313-1318.

Jia, H. P., T. Starner, M. Ackermann, P. Kirby, B. F. Tack, and P. B. J. McCray. 2001. Abundant human beta-defensin-1 expression in milk and mammary gland epithelium. J. Pediatr.138:109-112.

Kadowaki, N., S. Ho, S. Antonenko, R. W. Malefyt, R. A. Kastelein, F. Bazan, and Y. J. Liu. 2001. Subsets of human dendritic cell precursors express different toll-like receptors and respond to different microbial antigens. J. Exp. Med.194:863-869.

Klungland, H., A. Sabry, B. Heringstad, H. G. Olsen, H. Gomez-Raya, D. I. Vage, I. Olsaker, J. Odegard, G. Klemetsdal, N. Schulman, J. Vikki, J. Ruane, M. Aasland, K. Ronningen, and S. Lien. 2001. Quantitative trait loci affecting clinical mastitis and somatic cell count in dairy cattle. Mamm. Genome12:837-842.

Liu, A. Y., D. Destoumieux, A. V. Wong, C. H. Park, E. V. Valore, L. Liu, and T. Ganz. 2002. Human beta-defensin-2 production in keratinocytes is regulated by interleukin-1, bacteria, and the state of differentiation. J. Investig. Dermatol.118:275-281.

Mao, J., S. Marcos, S. K. Davis, J. Burzlaff, and H. M. Seyfert. 2001. Genomic distribution of three promoters of the bovine gene encoding acetyl-CoA carboxylase alpha and evidence that the nutritionally regulated promoter I contains a repressive element different from that in rat. Biochem. J.358:127-135.

Mao, J., A. J. Molenaar, T. T. Wheeler, and H.-M. Seyfert. 2002. Stat5 binding contributes to lactational stimulation of promoter III expressing the bovine acetyl-CoA-carboxylase α-encoding gene in the mammary gland. J. Mol. Endocrinol.29:73-88.

Medzhitov, R., P. Preston-Hurlburt, and C. A. Janeway, Jr. 1997. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature388:394-397.

Molenaar, A. J., S. R. Davis, and R. J. Wilkins. 1992. Expression of α-lactalbumin, α-S1-casein, and lactoferrin genes is heterogeneous in sheep and cattle mammary tissue. J. Histochem. Cytochem.40:611-618.

Moser, M., and K. M. Murphy. 2000. Dendritic cell regulation of TH1-TH2 development. Nat. Immunol.1:199-205.

O′Neil, D. A., E. M. Porter, D. Elewaut, G. M. Anderson, L. Eckmann, T. Ganz, and M. F. Kagnoff. 1999. Expression and regulation of the human beta-defensins hBD-1 and hBD-2 in intestinal epithelium. J. Immunol.163:6718-6724.

O′Neill, L. A. J. 2002. Toll-like receptor signal transduction and the tailoring of innate immunity: a role for Mal? Trends Immunol.23:296-300.

Ozinsky, A., D. M. Underhill, J. D. Fontenot, A. M. Hajjar, K. D. Smith, C. B. Wilson, L. Schroeder, and A. Aderem. 2000. The repertoire for pattern recognition of pathogens by the innate immune system is defined by cooperation between toll-like receptors. Proc. Natl. Acad. Sci. USA97:13766-13771.

Paape, M. J., K. Shafer-Weaver, A. V. Capuco, K. Van Oostveldt, and C. Burvenich. 2000. Immune surveillance of mammary tissue by phagocytic cells. Adv. Exp. Med. Biol.480:259-277.

Raghupathy, R. 1997. Th1-type immunity is incompatible with successful pregnancy. Immunol. Today18:478-482.

Riollet, C., P. Rainard, and B. Poutrel. 2000. Cells and cytokines in inflammatory secretions of bovine mammary gland. Adv. Exp. Med. Biol.480:247-258.

Rock, F. L., G. Hardiman, J. C. Timans, R. A. Kastelein, and J. F. Bazan. 1998. A family of human receptors structurally related to Drosophila Toll. Proc. Natl. Acad. Sci. USA95:588-593.

10.1074/jbc.M108830200

Schonwetter, B. S., E. D. Stolzenberg, and M. A. Zasloff. 1995. Epithelial antibiotics induced at sites of inflammation. Science267:1645-1648.

Schroder, J. M. 1999. Epithelial antimicrobial peptides: innate local host response elements. Cell. Mol. Life Sci.56:32-46.

Selstedt, M. E., Y.-Q. Tang, W. L. Morris, P. A. McGuire, M. J. Novotny, A. H. Henschen, and J. S. Cullor. 1993. Purification, primary structures, and antibacterial activities of β-defensins, a new family of antibacterial peptides from bovine neutrophils. J. Biol. Chem.268:6641-6648.

Shook, G. E. 1993. Genetic improvement of mastitis through selection on somatic cell count. Vet. Clin. N. Am.9:563-577.

Slonim, D., L. Kruglyak, L. Stein, and E. Lander. 1997. Building human genome maps with radiation hybrids. J. Comput. Biol.4:487-504.

Sordillo, L. M., and K. L. Streicher. 2002. Mammary gland immunity and mastitis susceptibility. J. Mammary Gland Biol. Neoplasia7:135-146.

Sparwasser, T., E. S. Koch, R. M. Vabulas, K. Heeg, G. B. Lipford, J. W. Ellwart, and H. Wagner. 1998. Bacterial DNA and immunostimulatory CpG oligonucleotides trigger maturation and activation of murine dendritic cells. Eur. J. Immunol.28:2045-2054.

Takeshita, F., C. A. Leifer, I. Gursel, K. J. Ishii, S. Takeshita, M. Gursel, and D. M. Klinman. 2001. Cutting edge: role of Toll-like receptor 9 in CpG DNA-induced activation of human cells. J. Immunol.167:3555-3558.

Tunzi, C. R., P. A. Harper, B. Bar-Oz, E. V. Valore, J. L. Semple, J. Watson-MacDonell, T. Ganz, and S. Ito. 2000. Beta-defensin expression in human mammary gland epithelia. Pediatr. Res.48:30-35.

Underhill, D. M., A. Ozinsky, A. M. Hajjar, A. Stevens, C. B. Wilson, M. Bassetti, and A. Aderem. 1999. The Toll-like receptor 2 is recruited to macrophage phagosomes and discriminates between pathogens. Nature401:811-815.

Wolfs, T. G., W. A. Buurman, A. van Schadewijk, B. de Vries, M. A. Daemen, P. S. Hiemstra, and C. van't Veer. 2002. In vivo expression of Toll-like receptor 2 and 4 by renal epithelial cells: IFN-gamma and TNF-alpha mediated up-regulation during inflammation. J. Immunol.168:1286-1293.

Womack, J. E., J. S. Johnson, E. K. Owens, C. E. Rexroad III, J. Schlapfer, and Y. P. Yang. 1997. A whole-genome radiation hybrid panel for bovine gene mapping. Mamm. Genome8:854-856.

Womack, J. E., and Y. D. Moll. 1986. Gene map of the cow: conservation of linkage with mouse and man. J. Hered.77:2-7.

Young B. and M. L. M. Anderson. 1985. Quantitative analysis of solution hybridisation p. 47-73. In B. D. Hames and S. J. Higgins (ed.) Nucleic acid hybridisation—a practical approach. IRL Press Oxford United Kingdom.

10.1038/415389a

Zhu, B., J. A. Smith, S. M. Tracey, B. A. Konfortov, K. Welzel, L. C. Schalkwyk, H. Lehrach, S. Kollers, J. Masabanda, J. Buitkamp, R. Fries, J. L. Williams, and J. R. Miller. 1999. A 5x genome coverage bovine BAC library: production, characterization, and distribution. Mamm. Genome10:706-709.

Thông tin
Thông tin xuất bản

American Society for Microbiology

Tập 11 Số 1

174-185

Thông tin tác giả