Human casein alpha s1 (CSN1S1) skews in vitro differentiation of monocytes towards macrophages

BMC Immunology

Matthias Schneider¹  Joachim Jose²  Ellen Bleck¹  Irina Altendorfer¹  Achim Braukmann²  Stefan Vordenbäumen¹ 

[1] Heinrich-Heine-University Düsseldorf, Department of Rheumatology, Moorenstr. 5, 40225 Düsseldorf, Germany;Westfälische Wilhelms-University Münster, Institute of Pharmaceutical and Medicinal Chemistry, Correnstr. 48, 48149 Münster, Germany

关键词: Differentiation;    Milk;    Interleukin-6;    Interleukin-1;    Inflammation;    Macrophage;   

Others  :  1077752
DOI  :  10.1186/1471-2172-14-46

 received in 2013-08-01, accepted in 2013-09-30,  发布年份 2013

【 摘 要 】

Background

The milk-derived protein human Casein alpha s1 (CSN1S1) has recently been detected in blood cells and was shown to possess proinflammatory properties. In the present study, we investigated the effect of CSN1S1 on the differentiation of monocytes.

Methods

Primary human monocytes were stimulated with recombinant CSN1S1 and compared to cells stimulated with GM-CSF/IL-4 or M-CSF/IFNγ. Morphological changes were assessed by microscopy and quantification of surface markers of differentiation by FACS analysis. Phagocytic activity of CSN1S1 stimulated cells was measured by quantification of zymosan labeled particle uptake. The role of mitogen activated protein kinases for CSN1S1-induced differentiation of monocytes and proinflammatory cytokine expression was assessed by supplementation of specific inhibitors.

Results

CSN1S1 at a concentration of 10 μg/ml resulted in morphological changes (irregular shape, pseudopodia) and aggregation of cells, comparable to changes observed in M-CSF/IFNγ differentiated macrophages. Surface marker expression was altered after 24 h with an upregulation of CD14 (mean 2.5 fold) and CD64 (1.9 fold) in CSN1S1 stimulated cells. CSN1S1 treated cells showed a characteristic surface marker pattern for macrophages after 120 h of incubation (CD14^high, CD64^high, CD83^low, CD1a^low) comparable to changes observed in M-CSF/IFNγ treated monocytes. Furthermore, phagocytic activity was increased 1.4 and 1.9 fold following stimulation with 10 μg/ml CSN1S1 after 24 and 48 h, respectively. Early GM-CSF, but not GM-CSF/IL-4 induced differentiation of monocytes towards dendritic cells (DC) was inhibited by addition of CSN1S1. Finally, CSN1S1 induced upregulation of CD14 was impeded by inhibition of ERK1/2, while inhibition of the mitogen activated protein kinases JNK and p38 did not influence cellular differentiation. However, JNK and p38 inhibitors impeded CSN1S1 induced secretion of the proinflammatory cytokines IL-1b or IL-6.

Conclusions

CSN1S1 skews in vitro differentiation of monocytes towards a macrophage-like phenotype. Data is accumulating that functions of CSN1S1 are beyond nutritional properties and include immunomodulatory effects.

【 授权许可】

   
2013 Vordenbäumen et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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【 参考文献 】

• [1]Hvatum M, Kanerud L, Hällgren R, Brandtzaeg P: The gut-joint axis: cross reactive food antibodies in rheumatoid arthritis. Gut 2006, 55:1240-1247.
• [2]Aihara K, Ishii H, Yoshida M: Casein-derived tripeptide, Val-Pro-Pro (VPP), modulates monocyte adhesion to vascular endothelium. J Arthroscler Thromb 2009, 16:594-603.
• [3]Katagiri T, Adachi I, Terao T, Osawa T: Alpha-casein-binding proteins of guinea pig macrophage membranes and their possible roles in chemotaxis. J Biochem 1980, 87:1421-1430.
• [4]Lewis SL, van Epps DE: Demonstration of specific receptors for fluoresceinated casein on human neutrophils and monocytes using flow cytometry. Inflammation 1983, 7:363-375.
• [5]Solymossy M, Nagy Z, Tóth G, Antoni F: Interaction of casein with human polymophonuclear cells. Biochem Med Metab Biol 1986, 35:193-301.
• [6]van Epps DE, Bankhurst AD, Williams RCJ: Casein-mediated neutrophil chemotaxis. Inflammation 1977, 2:115-123.
• [7]Procoví C, Conesa C, Barbana C, Pérez MD, Calvo M, Sánchez L: Comparison of the activity of human and bovine milk on two cell lines. J Dairy Res 2009, 76:308-316.
• [8]Ramos-Mandujano G, Weiss-Steider B, Melo B, Córdova Y, Ledesma-Martínez E, Bustos S, Silvestre O, Aguiniga I, Sosa N, Martínez I, et al.: Alpha-, beta- and kappa-caseins inhibit the proliferation of the myeloid cell lines 32D cl3 and WEHI-3 and exhibit different differentiation properties. Immunobiology 2008, 213:133-141.
• [9]Otaegui D, Mostafavi S, Bernard CCA, de Munain AL, Mousavi P, Oksenberg JR, Baranzini SE: Increased transcriptional activity of milk-related genes following the active phase of experimental autoimmune encephalomyeltitis and multiple sclerosis. J Immunol 2007, 179:4074-4082.
• [10]Galligan CL, Baig E, Bykerk V, Keystone EC, Fish EN: Distinctive gene expression signatures in rheumatoid arthritis synovial tissue fibroblast cells: correlates with disease activity. Genes Immunity 2007, 8:480-491.
• [11]Karlsson C, Dehne T, Lindahl A, Brittberg M, Pruss A, Sittinger M, Ringe J: Genome-wide expression profiling reveals new candidate genes associated with osteoarthritis. Osteoarthritis Cartilage 2010, 18:581-592.
• [12]Ungethuem U, Haeupl T, Witt H, Koczan D, Krenn V, Huber H, von Helversen TM, Drungowski M, Seyfert C, Zacher J, et al.: Molecular signature and nex candidates to target the pathogenesis of rheumatoid arthritis. Physiol Genomics 2010, 42:267-282.
• [13]Vordenbäumen S, Braukmann A, Petermann K, Scharf A, Bleck E, von Mikecz A, Jose J, Schneider M: Casein alpha s1 (CSN1S1) is expressed by human monocytes and upregulates the production of GM-CSF via p38 MAPK. J Immunol 2011, 186:592-601.
• [14]Amor S, Puentes F, Baker D, van der Valk P: Inflammation in neurodegenerative diseases. Immunology 2010, 129:154-169.
• [15]Gieseler R, Heise D, Soruri A, Schwartz P, Peters JH: In-vitro differentiation of mature dendritic cells from human blood monocytes. Development Immunol 1998, 6:25-39.
• [16]Werz O, Klemm J, Radmark O, Samuelsson B: p38 MAP kinase mediates stress-induced leukotriene synthesis in a human B-lymphocyte cell line. J Leukoc Biol 2001, 70:830-838.
• [17]Rose K, Finger IE, Ferenz KB: Interaction of clodornate with fibroblast growth factor 2 reduces FGF2-activity in endothelial cells. Biomed Pharmacother 2011, 65:46-51.
• [18]Chomarat P, Banchereau J, Davoust J, Palucka AK: IL-6 switches the differentiation of monocytes from dendritic cells to macrophages. Nat Immunol 2000, 1:510-514.
• [19]Netea MG, Lewis EC, Azam T, Joosten LAB, Jaekal J, Bae S-Y, Dinarello CA, Kim S-H: Interleukin-32 induces the differentiation of monocytes into macrophage-like cells. Proc Natl Acad Sci 2008, 105:3515-3520.
• [20]Krutzik SR, TAn B, Li H, Ochoa MT, Liu PT, Sharfstein SE, Graeber TG, Sieling PA, Liu Y-J, Rea TH, et al.: TLR activation triggers the rapid differentiation of monocytes into macrophages and dendritic cells. Nat Med 2005, 11:653-660.
• [21]Lo AS, Gorak-Stolinska P, Bachy V, Ibrahim MA, Kemeny DM, Maher J: Modulation of dendritic cell differentiation by colony-stimulating factor-1: role of phosphatidylinositol 3′-kinase and delayed caspase activation. J Leukoc Biol 2007, 82:1446-1454.
• [22]Hsu HY, Wen MH: Lipopolysaccharide-mediated reactive oxygen species and signal transduction in the regulation of Interleukin-1 gene expression. J Biol Chem 2002, 277:22131-22139.
• [23]Nobeli I, Favia AD, Thornton JM: Protein promiscuity and its implications for biotechnology. Nature Biotech 2009, 27:157-167.
• [24]Franco OL: Peptide promiscuity: an evolutionary concept for plant defense. FEBS Lett 2011, 585:995-1000.
• [25]Rijnkels M: Multispecies comparison of the casein gene loci and evolution of casein gene family. J Mammary Gland Biol Neoplasia 2002, 7:327-345.
• [26]Carrotta R, Canale C, Diaspro A, Trapani A, San Biagio PL, Bulone D: Inhibiting effect of alpha(S)1-casein on A beta(1–40) fibrillogenesis. Biochim Biophys Acta 1820, 2012:124-132.
• [27]Calder PC, Krauss-Etschmann S, de Jong EC, Dupont C, Frick J-S, Frokiaer H, Heinrich J, Garn H, Koletzko S, Lack G, et al.: Early nutrition and immunity - progress and perspectives. Br J Nutr 2006, 96:774-790.
• [28]Petermann K, Vordenbäumen S, Maas R, Braukmann A, Bleck E, Saenger T, Schneider M, Jose J: Autoantibodies to alpha(S)1-casein are induced by breast-feeding. PLoS One 2012, 7:e32716.
• [29]Barbouche R, Forveille M, Fischer A, Avrameas S, Durandy A: Spontaneous IgM autoantibody production in vitro by lymphocytes of normal human neonates. Scand J Immunol 1992, 35:659-667.
• [30]Kjeldsen F, Savitski MM, Nielsen ML, Shi L, Zubarev RA: On studying protein phosphorylation patterns using bottom-up LC-MS/MS: the case of human (alpha)-casein. Analyst 2007, 132:768-776.
• [31]Sallusto F, Lanzavecchia A: Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha. J Exp Med 1994, 179:109-1118.
• [32]Mebratu Y, Tesfaigzi Y: How ERK1/2 activation controls cell proliferation and cell death: is subcellular localization the answer? Cell Cycle 2009, 8:1168-1175.
• [33]Lönnerdal B, Forsum E: Casein content of human milk. Am J Clin Nutr 1985, 41:113-120.
• [34]Britton JB, Kastin AJ: Biologically active polypeptides in milk. Am J Med Sci 1991, 301:124-132.