IL-4 and IL-13 exposure during mucociliary differentiation of bronchial epithelial cells increases antimicrobial activity and expression of antimicrobial peptides

Respiratory Research

Pieter S Hiemstra³  Klaus F Rabe³  Sandra van Wetering¹  Frans A Prins²  Renate M Verhoosel³  Marianne AJA van Sterkenburg³  Jasmijn A Schrumpf³  Dennis K Ninaber³  Suzanne Zuyderduyn¹ 

[1] DCPrime BV, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands;Pathology, Leiden University Medical Center, Leiden, The Netherlands;Department of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands

关键词: inflammation;    allergy;    cell differentiation;    lung;    human;   

Others  :  796865
DOI  :  10.1186/1465-9921-12-59

 received in 2010-04-22, accepted in 2011-05-02,  发布年份 2011

【 摘 要 】

The airway epithelium forms a barrier against infection but also produces antimicrobial peptides (AMPs) and other inflammatory mediators to activate the immune system. It has been shown that in allergic disorders, Th2 cytokines may hamper the antimicrobial activity of the epithelium. However, the presence of Th2 cytokines also affects the composition of the epithelial layer which may alter its function. Therefore, we investigated whether exposure of human primary bronchial epithelial cells (PBEC) to Th2 cytokines during mucociliary differentiation affects expression of the human cathelicidin antimicrobial protein (hCAP18)/LL-37 and human beta defensins (hBD), and antimicrobial activity.

PBEC were cultured at an air-liquid interface (ALI) for two weeks in the presence of various concentrations of IL-4 or IL-13. Changes in differentiation and in expression of various AMPs and the antimicrobial proteinase inhibitors secretory leukocyte protease inhibitor (SLPI) and elafin were investigated as well as antimicrobial activity.

IL-4 and IL-13 increased mRNA expression of hCAP18/LL-37 and hBD-2. Dot blot analysis also showed an increase in hCAP18/LL-37 protein in apical washes of IL-4-treated ALI cultures, whereas Western Blot analysis showed expression of a protein of approximately 4.5 kDa in basal medium of IL-4-treated cultures. Using sandwich ELISA we found that also hBD-2 in apical washes was increased by both IL-4 and IL-13. SLPI and elafin levels were not affected by IL-4 or IL-13 at the mRNA or protein level. Apical wash obtained from IL-4- and IL-13-treated cultures displayed increased antimicrobial activity against Pseudomonas aeruginosa compared to medium-treated cultures. In addition, differentiation in the presence of Th2 cytokines resulted in increased MUC5AC production as has been shown previously.

These data suggest that prolonged exposure to Th2 cytokines during mucociliary differentiation contributes to antimicrobial defence by increasing the expression and release of selected antimicrobial peptides and mucus.

【 授权许可】

   
2011 Zuyderduyn et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Van Wetering S, van der Linden AC, van Sterkenburg MA, de Boer WI, Kuijpers AL, Schalkwijk J, et al.: Regulation of SLPI and elafin release from bronchial epithelial cells by neutrophil defensins. Am J Physiol Lung Cell Mol Physiol 2000, 278:L51-L58.
• [2]Williams SE, Brown TI, Roghanian A, Sallenave JM: SLPI and elafin: one glove, many fingers. Clin Sci (Lond) 2006, 110:21-35.
• [3]Hiemstra PS, Maassen RJ, Stolk J, Heinzel-Wieland R, Steffens GJ, Dijkman JH: Antibacterial activity of antileukoprotease. Infect Immun 1996, 64:4520-4524.
• [4]Simpson AJ, Wallace WA, Marsden ME, Govan JR, Porteous DJ, Haslett C, et al.: Adenoviral augmentation of elafin protects the lung against acute injury mediated by activated neutrophils and bacterial infection. J Immunol 2001, 167:1778-1786.
• [5]Sallenave JM: Antimicrobial activity of antiproteinases. Biochem Soc Trans 2002, 30:111-115.
• [6]McNeely TB, Shugars DC, Rosendahl M, Tucker C, Eisenberg SP, Wahl SM: Inhibition of human immunodeficiency virus type 1 infectivity by secretory leukocyte protease inhibitor occurs prior to viral reverse transcription. Blood 1997, 90:1141-1149.
• [7]Del Prete GF, De Carli M, D'Elios MM, Maestrelli P, Ricci M, Fabbri L, et al.: Allergen exposure induces the activation of allergen-specific Th2 cells in the airway mucosa of patients with allergic respiratory disorders. Eur J Immunol 1993, 23:1445-1449.
• [8]Robinson DS, Hamid Q, Ying S, Tsicopoulos A, Barkans J, Bentley AM, et al.: Predominant TH2-like bronchoalveolar T-lymphocyte population in atopic asthma. N Engl J Med 1992, 326:298-304.
• [9]Van Wetering S, Zuyderduyn S, Ninaber DK, van Sterkenburg MA, Rabe KF, Hiemstra PS: Epithelial differentiation is a determinant in the production of eotaxin-2 and -3 by bronchial epithelial cells in response to IL-4 and IL-13. Mol Immunol 2007, 44:803-811.
• [10]Zhu Z, Homer RJ, Wang Z, Chen Q, Geba GP, Wang J, et al.: Pulmonary expression of interleukin-13 causes inflammation, mucus hypersecretion, subepithelial fibrosis, physiologic abnormalities, and eotaxin production. J Clin Invest 1999, 103:779-788.
• [11]Laoukili J, Perret E, Willems T, Minty A, Parthoens E, Houcine O, et al.: IL-13 alters mucociliary differentiation and ciliary beating of human respiratory epithelial cells. J Clin Invest 2001, 108:1817-1824.
• [12]Yoshisue H, Puddicombe SM, Wilson SJ, Haitchi HM, Powell RM, Wilson DI, et al.: Characterization of ciliated bronchial epithelium 1, a ciliated cell-associated gene induced during mucociliary differentiation. Am J Respir Cell Mol Biol 2004, 31:491-500.
• [13]Atherton HC, Jones G, Danahay H: IL-13-induced changes in the goblet cell density of human bronchial epithelial cell cultures: MAP kinase and phosphatidylinositol 3-kinase regulation. Am J Physiol Lung Cell Mol Physiol 2003, 285:L730-L739.
• [14]Ong PY, Ohtake T, Brandt C, Strickland I, Boguniewicz M, Ganz T, et al.: Endogenous antimicrobial peptides and skin infections in atopic dermatitis. N Engl J Med 2002, 347:1151-1160.
• [15]Nomura I, Goleva E, Howell MD, Hamid QA, Ong PY, Hall CF, et al.: Cytokine milieu of atopic dermatitis, as compared to psoriasis, skin prevents induction of innate immune response genes. J Immunol 2003, 171:3262-3269.
• [16]Albanesi C, Fairchild HR, Madonna S, Scarponi C, De Pita O, Leung DY, et al.: IL-4 and IL-13 negatively regulate TNF-alpha- and IFN-gamma-induced beta-defensin expression through STAT-6, suppressor of cytokine signaling (SOCS)-1, and SOCS-3. J Immunol 2007, 179:984-992.
• [17]Howell MD, Gallo RL, Boguniewicz M, Jones JF, Wong C, Streib JE, et al.: Cytokine milieu of atopic dermatitis skin subverts the innate immune response to vaccinia virus. Immunity 2006, 24:341-348.
• [18]Beisswenger C, Kandler K, Hess C, Garn H, Felgentreff K, Wegmann M, et al.: Allergic airway inflammation inhibits pulmonary antibacterial host defense. J Immunol 2006, 177:1833-1837.
• [19]Van Wetering S, van der Linden AC, van Sterkenburg MA, Rabe KF, Schalkwijk J, Hiemstra PS: Regulation of secretory leukocyte proteinase inhibitor (SLPI) production by human bronchial epithelial cells: increase of cell-associated SLPI by neutrophil elastase. J Investig Med 2000, 48:359-366.
• [20]Tjabringa GS, Vos JB, Olthuis D, Ninaber DK, Rabe KF, Schalkwijk J, et al.: Host defense effector molecules in mucosal secretions. FEMS Immunol Med Microbiol 2005, 45:151-158.
• [21]Wingens M, van Bergen BH, Hiemstra PS, Meis JF, Vlijmen-Willems IM, Zeeuwen PL, et al.: Induction of SLPI (ALP/HUSI-I) in epidermal keratinocytes. J Invest Dermatol 1998, 111:996-1002.
• [22]Proud D, Sanders SP, Wiehler S: Human rhinovirus infection induces airway epithelial cell production of human beta-defensin 2 both in vitro and in vivo. J Immunol 2004, 172:4637-4645.
• [23]Diamond G, Yim S, Rigo I, McMahon L: Measuring antimicrobial peptide activity on epithelial surfaces in cell culture. Methods Mol Biol 2010, 618:371-382.
• [24]Murakami M, Lopez-Garcia B, Braff M, Dorschner RA, Gallo RL: Postsecretory processing generates multiple cathelicidins for enhanced topical antimicrobial defense. J Immunol 2004, 172:3070-3077.
• [25]Peyret-Lacombe A, Duplan H, Watts M, Charveron M, Brunel G: Antimicrobial peptide modulation in a differentiated reconstructed gingival epithelium. Cell Tissue Res 2007, 328:85-95.
• [26]Hase K, Eckmann L, Leopard JD, Varki N, Kagnoff MF: Cell differentiation is a key determinant of cathelicidin LL-37/human cationic antimicrobial protein 18 expression by human colon epithelium. Infect Immun 2002, 70:953-963.
• [27]Kikuchi T, Shively JD, Foley JS, Drazen JM, Tschumperlin DJ: Differentiation-dependent responsiveness of bronchial epithelial cells to IL-4/13 stimulation. Am J Physiol Lung Cell Mol Physiol 2004, 287:L119-L126.
• [28]Singh PK, Tack BF, McCray PB, Welsh MJ: Synergistic and additive killing by antimicrobial factors found in human airway surface liquid. Am J Physiol Lung Cell Mol Physiol 2000, 279:L799-L805.
• [29]Harder J, Meyer-Hoffert U, Teran LM, Schwichtenberg L, Bartels J, Maune S, et al.: Mucoid Pseudomonas aeruginosa, TNF-alpha, and IL-1beta, but Not IL-6, Induce Human beta -Defensin-2 in Respiratory Epithelia. Am J Respir Cell Mol Biol 2000, 22:714-721.
• [30]Bals R, Wang X, Zasloff M, Wilson JM: The peptide antibiotic LL-37/hCAP-18 is expressed in epithelia of the human lung where it has broad antimicrobial activity at the airway surface. Proc Natl Acad Sci USA 1998, 95:9541-9546.
• [31]Gray TE, Guzman K, Davis CW, Abdullah LH, Nettesheim P: Mucociliary differentiation of serially passaged normal human tracheobronchial epithelial cells. Am J Respir Cell Mol Biol 1996, 14:104-112.
• [32]Vermeer PD, Harson R, Einwalter LA, Moninger T, Zabner J: Interleukin-9 induces goblet cell hyperplasia during repair of human airway epithelia. Am J Respir Cell Mol Biol 2003, 28:286-295.
• [33]Felgentreff K, Beisswenger C, Griese M, Gulder T, Bringmann G, Bals R: The antimicrobial peptide cathelicidin interacts with airway mucus. Peptides 2006, 27:3100-3106.
• [34]Bucki R, Byfield FJ, Janmey PA: Release of the antimicrobial peptide LL-37 from DNA/F-actin bundles in cystic fibrosis sputum. Eur Respir J 2007, 29:624-632.
• [35]Davidson DJ, Currie AJ, Reid GS, Bowdish DM, MacDonald KL, Ma RC, et al.: The cationic antimicrobial peptide LL-37 modulates dendritic cell differentiation and dendritic cell-induced T cell polarization. J Immunol 2004, 172:1146-1156.