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

Barry A Gusterson²  Gavin P Sandilands³  Roderick K Ferrier²  Alexandra K Bell²  Victoria J Heath²  Marie-Anne Duffy²  Stephen J Weber-Hall¹  Claire R Davies²  Joanna S Morris²  Torsten Stein² 

[1] Abbott Laboratories Limited, Diagnostic Division, Maidenhead, Berkshire, UK;Division of Cancer Sciences and Molecular Pathology, Western Infirmary, Glasgow, UK;Department of Pathology, Western Infirmary, Glasgow, UK

关键词: microarray;    mammary gland;    involution;    acute-phase response;   

 received in 2003-09-16, accepted in 2003-11-21,  发布年份 2003

【 摘 要 】

Introduction

Involution of the mammary gland is a complex process of controlled apoptosis and tissue remodelling. The aim of the project was to identify genes that are specifically involved in this process.

Methods

We used Affymetrix oligonucleotide microarrays to perform a detailed transcript analysis on the mechanism of controlled involution after withdrawal of the pups at day seven of lactation. Some of the results were confirmed by semi-quantitative reverse transcriptase polymerase chain reaction, Western blotting or immunohistochemistry.

Results

We identified 145 genes that were specifically upregulated during the first 4 days of involution; of these, 49 encoded immunoglobulin genes. A further 12 genes, including those encoding the signal transducer and activator of transcription 3 (STAT3), the lipopolysaccharide receptor (CD14) and lipopolysaccharide-binding protein (LBP), were involved in the acute-phase response, demonstrating that the expression of acute-phase response genes can occur in the mammary gland itself and not only in the liver. Expression of LBP and CD14 was upregulated, at both the RNA and protein level, immediately after pup withdrawal; CD14 was strongly expressed in the luminal epithelial cells. Other genes identified suggested neutrophil activation early in involution, followed by macrophage activation late in the process. Immunohistochemistry and histological staining confirmed the infiltration of the involuting mammary tissue with neutrophils, plasma cells, macrophages and eosinophils.

Conclusion

Oligonucleotide microarrays are a useful tool for identifying genes that are involved in the complex developmental process of mammary gland involution. The genes identified are consistent with an immune cascade, with an early acute-phase response that occurs in the mammary gland itself and resembles a wound healing process.

【 授权许可】

   
2004 Stein et al., licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.

【 图 表 】

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【 参考文献 】

• [1]Richert MM, Schwertfeger KL, Ryder JW, Anderson SM: An atlas of mouse mammary gland development. J Mammary Gland Biol Neoplasia 2000, 5:227-241.
• [2]Masso-Welch PA, Darcy KM, Stangle-Castor NC, Ip MM: A developmental atlas of rat mammary gland histology. J Mammary Gland Biol Neoplasia 2000, 5:165-185.
• [3]Lund LR, Romer J, Thomasset N, Solberg H, Pyke C, Bissell MJ, Dano K, Werb Z: Two distinct phases of apoptosis in mammary gland involution: proteinase-independent and -dependent pathways. Development 1996, 122:181-193.
• [4]Li M, Liu X, Robinson G, Bar-Paled U, Wagner K-U, Young WS, Hennighausen L, Furth PA: Mammary derived signals activate programmed cell during the involuting mammary gland. Proc Natl Acad Sci USA 1997, 94:3425-3430.
• [5]Marti A, Lazar H, Ritter P, Jaggi R: Transcription factor activities and gene expression during mouse mammary gland involution. J Mammary Gland Biol Neoplasia 1999, 4:145-152.
• [6]Master SR, Hartman JL, D'Cruz CM, Moody SE, Keiper EA, Ha SI, Cox JD, Belka GK, Chodosh LA: Functional microarray analysis of mammary organogenesis reveals a developmental role in adaptive thermogenesis. Mol Endocrinol 2000, 16:1185-1203.
• [7]Clarkson RWE, Wayland MT, Lee J, Freeman T, Watson CJ: Gene expression profiling of mammary gland development reveals putative roles for death receptors and immune mediators in post-lactational regression. Breast Cancer Res 2004, 6:R92-R109. BioMed Central Full Text
• [8]Eckersall PD: Recent advances and future prospects for the use of acute phase proteins as markers of disease in animals. Revue Méd Vét 2000, 151:577-584.
• [9]Dinarello CA: Interleukin-1 and the pathogenesis of the acute phase response. New Engl J Med 1984, 311:1413-1418.
• [10]Heinrich PC, Castell JC, Andus T: Interleukin-6 and the acute phase response. Biochem J 1990, 265:621-636.
• [11]Beutler B, Cerami A: Cachectin/tumor necrosis factor: an endogenous mediator of shock and inflammation. Immunol Res 1986, 5:281-293.
• [12]Anderson JC: The increased resistance of mice to experimental staphylococcal mastitis following inoculation of endotoxin. Res Vet Sci 1976, 21:64-68.
• [13]Chapman RS, Lourenco PC, Tonner E, Flint DJ, Selbert S, Takeda K, Akira S, Clarke AR, Watson CJ: Suppression of epithelial apoptosis and delayed mammary gland involution in mice with a conditional knockout of Stat3. Genes Dev 1999, 13:2604-2616.
• [14]Humphreys RC, Bierie B, Zhao L, Raz R, Levy D, Hennighausen L: Deletion of Stat3 blocks mammary gland involution and extends functional competence of the secretory epithelium in the absence of lactogenic stimuli. Endocrinology 2002, 143:3641-3650.
• [15]Akira S, Nishio Y, Inoue M, Wang XJ, Wei S, Matsusaka T, Yoshida K, Sudo T, Naruta M, Kishimoto T: Molecular cloning of APRF, a novel IFN-stimulated gene factor 3 p91-related transcription factor involved in the gp130-mediated signaling pathway. Cell 1994, 77:63-71.
• [16]Zhong Z, Wen Z, Darnell JE: Stat3: a new family member that is activated through tyrosine phosphorylation in response to EGF and IL-6. Science 1994, 264:95-98.
• [17]Trautwein C, Rakemann T, Niehof M, Rose-John S, Manns MP: Acute-phase response factor, increased binding, and target gene transcription during liver regeneration. Gastroenterology 1996, 110:1854-1862.
• [18]Schumann RR, Kirschning CJ, Unbehaun A, Aberle HP, Knope HP, Lamping N, Ulevitch RJ, Herrmann F: The lipopolysaccharide-binding protein is a secretory class 1 acute-phase protein whose gene is transcriptionally activated by APRF/STAT-3 and other cytokine-inducible nuclear proteins. Mol Cell Biol 1996, 16:3490-3503.
• [19]Cantwell CA, Sterneck E, Johnson PF: Interleukin-6-specific activation of the C/EBPdelta gene in hepatocytes is mediated by Stat3 and Sp1. Mol Cell Biol 1998, 18:2108-2117.
• [20]Boudreau F, Yu SJ, Asselin C: CCAAT/enhancer binding proteins beta and delta regulate alpha1-acid glycoprotein gene expression in rat intestinal epithelial cells. DNA Cell Biol 1998, 17:669-677.
• [21]Alam T, An MR, Papaconstantinou J: Differential expression of three C/EBP isoforms in multiple tissues during the acute phase response. J Biol Chem 1992, 267:5021-5024.
• [22]Pan Z, Hetherington CJ, Zhang DE: CCAAT/enhancer-binding protein activates the CD14 promoter and mediates transforming growth factor beta signaling in monocyte development. J Biol Chem 1999, 274:23242-23248.
• [23]Devitt A, Moffa OD, Raykundalia C, Capra JD, Simmons DL, Gregory CD: Human CD14 mediates recognition and phagocytosis of apoptotic cells. Nature 1998, 392:505-509.
• [24]Wright SD: CD14: a leukocyte membrane protein that functions in the response to endotoxin [abstract]. FASEB J 1990, 4:A1848.
• [25]Wright SD, Ramos RA, Tobias PS, Ulevitch RJ, Mathison JC: CD14, a receptor for complexes of lipopolysaccharide (LPS) and LPS binding protein. Science 1990, 249:1431-1433.
• [26]Paape MJ, Lillius EM, Wiitanen PA, Kontio MP: Intramammary defense against infections induced by Escherichia coli in cows. Am J Vet Res 1996, 57:477-482.
• [27]Labeta MO, Vidal K, Nores JE, Arias M, Vita N, Morgan BP, Guillemot JC, Loyaux D, Ferrara P, Schmid D, Affolter M, Borysiewicz LK, Donnet-Hughes A, Schiffrin EJ: Innate recognition of bacteria in human milk is mediated by a milk-derived highly expressed pattern recognition receptor, soluble CD14. J Exp Med 2000, 191:1807-1812.
• [28]Walker NI, Bennett RE, Kerr JFR: Cell death by apoptosis during involution of the lactating breast in mice and rats. Am J Anat 1989, 185:19-32.
• [29]Fadok VA: Clearance: the last and often forgotten stage of apoptosis. J Mammary Gland Biol Neoplasia 1999, 4:203-211.
• [30]Monks J, Geske FJ, Lehman L, Fadok VA: Do inflammatory cells participate in mammary gland involution? J Mammary Gland Biol Neoplasia 2002, 7:163-176.
• [31]Gregory CD: CD14-dependent clearance of apoptotic cells: relevance to the immune system. Curr Opin Immunol 2000, 12:27-34.
• [32]Old LJ: Tumor necrosis factor. Sci Am 1988, 258:59-75.
• [33]Fadok VA, Bratton DL, Konowal A, Freed PW, Westcott JY, Henson PM: Macrophages that have ingested apoptotic cells in vitro inhibit proinflammatory cytokine production through autocrine/paracrinemechanisms involving TGF β, PGE2, and PAF. J Clin Invest 1998, 1001:890-898.
• [34]Laurent PE: Clinical measurement of acute phase proteins to detect and monitor infectious disease. In In Acute Phase Proteins in the Acute Phase Response. Edited by Pepys MB. New York: Springer-Verlag; 1989:151-159.
• [35]Jin FY, Nathan C, Radzioch D, Ding A: Secretory leukocyte protease inhibitor: a macrophage product induced by and antagonistic to bacterial lipopolysaccharide. Cell 1997, 88:417-426.
• [36]Zhu J, Nathan C, Jin W, Sim D, Ashcroft GS, Wahl SM, Lacomis L, Erdjument-Bromage H, Tempst P, Wright CD, Ding A: Conversion of proepithelin to epithelins. Roles of SLPI and elastase in host defense and wound repair. Cell 2002, 111:867-878.
• [37]Lutticken C, Wegenka UM, Yuan J, Buschmann J, Schindler C, Ziemiecki A, Harpur AG, Wilks AF, Yasukawa K, Taga T: Association of transcription factor APRF and protein kinase Jak1 with the interleukin-6 signal transducer gp130. Science 1994, 263:89-92.
• [38]Hutt JA, DeWille JW: Oncostatin M induces growth arrest of mammary epithelium via a CCAAT/enhancer-binding protein delta-dependent pathway. Mol Cancer Ther 2002, 1:601-610.
• [39]Grant SL, Douglas AM, Goss GA, Begley CG: Oncostatin M and leukemia inhibitory factor regulate the growth of normal human breast epithelial cells. Growth Factors 2001, 19:153-162.
• [40]Schere-Levy C, Buggiano V, Quaglino A, Gattelli A, Cirio MC, Piazzon I, Vanzulli S, Kordon EC: Leukemia inhibitory factor induces apoptosis of the mammary epithelial cells and participates in mouse mammary gland involution. Exp Cell Res 2003, 282:35-47.
• [41]Kritikou EA, Sharkey A, Abell K, Came PJ, Anderson E, Clarkson RWE, Watson CJ: A dual, non-redundant, role for LIF as a regulator of development and STAT3-mediated cell death in mammary gland. Development 2003, 130:3459-3468.
• [42]Lee CS, McDowell GH, Lascelles AK: The importance of macrophages in the removal of fat from the involuting mammary gland. Res Vet Sci 1969, 10:34-38.
• [43]Nordin W, Lee CS: Cytology of milk in guinea pigs. Acta Anat 1982, 113:135-144.
• [44]Lee CS, McCauley I, Hartmann PE: Light and electron microscopy of cells in pig colostrum, milk and involution secretion. Acta Anat 1983, 116:126-135.
• [45]Colditz IG: Studies on the inflammatory response during involution of the ovine mammary gland. Q J Exp Physiol 1988, 73:363-368.
• [46]O'Donnell LC, Druhan LJ, Avalos BR: Molecular characterization and expression analysis of leucine-rich alpha2-glycoprotein, a novel marker of granulocytic differentiation. J Leukoc Biol 2002, 72:478-485.
• [47]Engelhardt E, Toksoy A, Goebeler M, Debus S, Brocker EB, Gillitzer R: Chemokines IL-8, GROalpha, MCP-1, IP-10, and Mig are sequentially and differentially expressed during phase-specific infiltration of leukocyte subsets in human wound healing. Am J Pathol 1998, 153:1849-1860.
• [48]Wiekowski MT, Chen SC, Zalamea P, Wilburn BP, Kinsley DJ, Sharif WW, Jensen KK, Hedrick JA, Manfra D, Lira SA: Disruption of neutrophil migration in a conditional transgenic model: evidence for CXCR2 desensitization in vivo. J Immunol 2001, 167:7102-7110.
• [49]Aubry F, Habasque C, Satie AP, Jegou B, Samson M: Expression and regulation of the CXC-chemokines, GRO/KC and IP-10/ mob-1 in rat seminiferous tubules. Eur Cytokine Netw 2000, 11:690-698.
• [50]Heeckeren A, Walenga R, Konstan MW, Bonfield T, Davis PB, Ferkol T: Excessive inflammatory response of cystic fibrosis mice to bronchopulmonary infection with Pseudomonas aeruginosa. J Clin Invest 1997, 100:2810-2815.
• [51]Jones CE, Chan K: Interleukin-17 stimulates the expression of interleukin-8, growth-related oncogene-alpha, and granulocyte-colony-stimulating factor by human airway epithelial cells. Am J Respir Cell Mol Biol 2002, 26:748-753.
• [52]Mehrad B, Strieter RM, Moore TA, Tsai WC, Lira SA, Standiford TJ: CXC chemokine receptor-2 ligands are necessary components of neutrophil-mediated host defense in invasive pulmonary aspergillosis. J Immunol 1999, 163:6086-6094.
• [53]Endlich B, Armstrong D, Brodsky J, Novotny M, Hamilton TA: Distinct temporal patterns of macrophage-inflammatory protein-2 and KC chemokine gene expression in surgical injury. J Immunol 2002, 168:3586-3594.
• [54]Rovai LE, Herschman HR, Smith JB: The murine neutrophil-chemoattractant chemokines LIX, KC, and MIP-2 have distinct induction kinetics, tissue distributions, and tissue-specific sensitivities to glucocorticoid regulation in endotoxemia. J Leukoc Biol 1998, 64:494-502.
• [55]Anisowicz A, Zajchowski D, Stenman G, Sager R: Functional diversity of GRO gene expression in human fibroblasts and mammary epithelial cells. Proc Nat Acad Sci USA 1988, 85:9645-9649.
• [56]Paape M, Mehrzad J, Zhao J, Detilleux J, Burvenich C: Defense of the bovine mammary gland by polymorphonuclear neutrophil leukocytes. J Mammary Gland Biol Neoplasia 2002, 7:109-121.
• [57]Gouon-Evans V, Rothenberg ME, Pollard JW: Postnatal mammary gland development requires macrophages and eosinophils. Development 2000, 127:2269-2282.
• [58]Sleeman MA, Fraser JK, Murison JG, Kelly SL, Prestidge RL, Palmer DJ, Watson JD, Kumble KD: B cell- and monocyte-activating chemokine (BMAC), a novel non-ELR alpha-chemokine. Int Immunol 2000, 12:677-689.
• [59]Kurth I, Willimann K, Schaerli P, Hunziker T, Clark-Lewis I, Moser B: Monocyte selectivity and tissue localization suggests a role for breast and kidney-expressed chemokine (BRAK) in macrophage development. J Exp Med 2001, 194:855-861.
• [60]Nickerson SC: Immunological aspects of mammary involution. J Dairy Sci 1989, 72:1665-1678.
• [61]Myokai F, Takashiba S, Lebo R, Amar S: A novel lipopolysaccharide-induced transcription factor regulating tumor necrosis factor α gene expression: molecular cloning, sequencing, characterization, and chromosomal assignment. Proc Natl Acad Sci USA 1999, 96:4518-4523.
• [62]Kushner I, Mackiewicz A: Acute phase proteins as disease markers. Dis Markers 1987, 5:1-11.
• [63]Hochepied T, Berger FG, Baumann H, Libert C: α1-acid glycoprotein: and acute phase protein with inflammatory and immunomodulating properties. Cyt Growth Factor Rev 2003, 14:25-34.
• [64]Gitlin JD: Transcriptional regulation of ceruloplasmin gene expression during inflammation. J Biol Chem 1988, 263:6281-6287.
• [65]Pietzsch A, Buchler C, Aslanidis C, Schmitz G: Identification and characterization of a novel monocyte/macrophage differentiation-dependent gene that is responsive to lipopolysaccharide, ceramide, and lysophosphatidylcholine. Biochem Biophys Res Commun 1997, 235:4-9.