Tissue-specific human beta-defensins (HBD)-1, HBD-2 and HBD-3 secretion profile from human amniochorionic membranes stimulated with Candida albicans in a two-compartment tissue culture system
During intrauterine infection, amniochorionic membranes represent a mechanical and immunological barrier against dissemination of infection. Human beta defensins (HBD)-1, HBD-2, and HBD-3 are key elements of innate immunity that represent the first line of defense against different pathogen microorganisms associated with preterm labor. The aim of this work was to characterize the individual contribution of the amnion (AMN) and choriodecidua (CHD) regions to the secretion of HBD-1, HBD-2 and HBD-3, after stimulation with Candida albicans.
Full-thickness human amniochorionic membranes were obtained after delivery by elective cesarean section from women at 37-40 wk of gestation with no evidence of active labor. The membranes were cultured in a two-compartment experimental model in which the upper compartment is delimited by the amnion and the lower chamber by the choriodecidual membrane. One million of Candida albicans were added to either the AMN or the CHD face or to both and compartmentalized secretion profiles of HBD-1, HBD-2, and HBD-3 were quantified by ELISA. Tissue immunolocalization was performed to detect the presence of HBD-1, -2, -3 in tissue sections stimulated with Candida albicans.
HBD-1 secretion level by the CHD compartment increased 2.6 times (27.30 [20.9-38.25] pg/micrograms protein) when the stimulus with Candida albicans was applied only on this side of the membrane and 2.4 times (26.55 [19.4-42.5] pg/micrograms protein) when applied to both compartments simultaneously. HBD-1 in the amniotic compartment remained without significant changes. HBD-2 secretion level increased significantly in the CHD when the stimulus was applied only to this region (2.49 [1.49-2.95] pg/micrograms protein) and simultaneously to both compartments (2.14 [1.67- 2.91] pg/micrograms protein). When the stimulus was done in the amniotic compartment HBD-2 remained without significant changes in both compartments. HBD-3 remained without significant changes in both compartments regardless of the stimulation modality. Localization of immune-reactive forms of HBD-1, HBD-2, and HBD-3 was carried out by immunohistochemistry confirming the cellular origin of these peptides.
Selective stimulation of amniochorionic membranes with Candida albicans resulted in tissue-specific secretion of HBD-1 and HBD-2, mainly in the CHD, which is the first region to become infected during an ascending infection.
- Levy O: Innate immunity of the newborn: basic mechanisms and clinical correlates. Nat Rev Immunol 2007, 7:379–390. CrossRef
- Yoon BH, Romero R, Moon JB, Shim SS, Kim M, Kim G, Jun JK: Clinical significance of intra-amniotic inflammation in patients with preterm labor and intact membranes. Am J Obstet Gynecol 2001, 185:1130–1136. CrossRef
- Asrat T: Intra-amniotic infection in patients with preterm prelabor rupture of membranes. Pathophysiology, Detection, and Management. Clinic Perinatol 2001, 28:735–751. CrossRef
- Romero R, Mazor M: Infection and preterm labor. Clin Obstet Gynecol 1988, 31:553–584. CrossRef
- Parry S, Strauus JF: Premature rupture of the fetal membranes. N Engl J Med 1998, 338:663–668. CrossRef
- Malak TM, Ockleford CD, Bell SC, Dalgleish R, Bright N, Macvicar J: Confocal immunofluorescence localization of collagen types I, III, IV, V and VI and their ultrastructural organization in term human fetal membranes. Placenta 1993, 14:385–406. CrossRef
- Klein LL, Gibbs RS: Infection and preterm birth. Obstet Gynecol Clin North Am 2005, 32:397–410. CrossRef
- Talmi YP, Sigler L, Inge E, Finkelstein Y, Zohar Y: Antibacterial properties of human amniotic membranes. Placenta 199, 12:285–288.
- Kjaergaard N, Hein M, Hyttel L, Helmig RB, Schønheyder HC, Uldbjerg N, Madsen H: Antibacterial properties of human amnion and chorion in vitro. Eur J Obstet Gynecol Reprod Biol 2001, 94:224–229. CrossRef
- King AE, Paltoo A, Kelly RW, Sallenave JM, Bocking AD, Challis JR: Expression of natural antimicrobials by human placenta and fetal membranes. Placenta 2007, 28:161–169. CrossRef
- Hancock RE, Diamond G: The role of cationic antimicrobial peptides in innate host defences. Trends Microbiol 2000, 8:402–410. CrossRef
- De Smet K, Contreras R: Human antimicrobial peptides: defensins, cathelicidins and histatins. Biotechnol Lett 2005, 27:1337–1347. CrossRef
- Yeaman MR, Yount NY: Mechanisms of antimicrobial peptide action and resistance. Pharmacol Rev 2003, 55:27–55. CrossRef
- Ganz T, Selsted ME, Szklarek D, Harwig SS, Daher K, Bainton DF, Lehrer RI: Defensins. Natural peptide antibiotics of human neutrophils. J Clin Invest 1985, 76:1427–1435. CrossRef
- Schneider JJ, Unholzer A, Schaller M, Schäfer-Korting M, Korting HC: Human defensins. J Mol Med (Berl) 2005, 83:587–595. CrossRef
- Stock SJ, Kelly RW, Riley SC, Calder AA: Natural antimicrobial production by the amnion. Am J Obstet Gynecol 2007, 196:255.e1–255.e6. CrossRef
- Buhimschi IA, Jabr M, Buhimschi CS, Petkova AP, Weiner CP, Saed GM: The novel antimicrobial peptide beta3-defensin is produced by the amnion: a possible role of the fetal membranes in innate immunity of the amniotic cavity. Am J Obstet Gynecol 2004, 191:1678–1687. CrossRef
- Szukiewicz D, Szewczyk G, Pyzlak M, Klimkiewicz J, Maslinska D: Increased production of beta-defensin 3 (hBD-3) by human amniotic epithelial cells (HAEC) after activation of toll-like receptor 4 in chorioamnionitis. Inflamm Res 2008,57(Suppl 1):S67-S68. CrossRef
- Soto E, Espinoza J, Nien JK, Kusanovic JP, Erez O, Richani K, Santolaya-Forgas J, Romero R: Human beta-defensin-2: a natural antimicrobial peptide present in amniotic fluid participates in the host response to microbial invasion of the amniotic cavity. J Matern Fetal Neonatal Med 2007, 20:15–22. CrossRef
- Espinoza J, Chaiworapongsa T, Romero R, Edwin S, Rathnasabapathy C, Gomez R, Bujold E, Camacho N, Kim YM, Hassan S, Blackwell S, Whitty J, Berman S, Redman M, Yoon BH, Sorokin Y: Antimicrobial peptides in amniotic fluid: defensins, calprotectin and bacterial/permeability-increasing protein in patients with microbial invasion of the amniotic cavity, intra-amniotic inflammation, preterm labor and premature rupture of membranes. J Matern Fetal Neonatal Med 2003, 13:2–21. CrossRef
- Erez O, Romero R, Tarca AL, Chaiworapongsa T, Kim YM, Than NG, Vaisbuch E, Draghici S, Tromp G: Differential expression pattern of genes encoding for anti-microbial peptides in the fetal membranes of patients with spontaneous preterm labor and intact membranes and those with preterm prelabor rupture of the membranes. J Matern Fetal Neonatal Med 2009, 22:1103–1115. CrossRef
- King AE, Kelly RW, Sallenave JM, Bocking AD, Challis JR: Innate immune defences in the human uterus during pregnancy. Placenta 2007, 28:1099–1106. CrossRef
- Zaga-Clavellina V, Garcia-Lopez G, Flores-Espinosa P: Evidence of in vitro differential secretion of human beta-defensins-1, -2, and -3 after selective exposure to Streptococcus agalactiae in human fetal membranes. J Matern Fetal Neonatal Med 2012, 4:258–263.
- Zaga-Clavellina V, Martha RV, Flores-Espinosa P: In vitro secretion profile of pro-inflammatory cytokines IL-1β, TNF-α, IL-6, and of human beta-defensins (HBD)-1, HBD-2, and HBD-3 from human chorioamniotic membranes after selective stimulation with Gardnerella vaginalis. Am J Reprod Immunol 2012, 67:34–43. CrossRef
- Garcia-Lopez G, Flores-Espinosa P, Zaga-Clavellina V: Tissue-specific human beta-defensins (HBD)1, HBD2, and HBD3 secretion from human extra-placental membranes stimulated with Escherichia coli. Reprod Biol Endocrinol 2010, 8:146. CrossRef
- Donders GG, Moerman P, Caudron J, Van Assche FA: Intra-uterine Candida infection: a report of four infected fetusses from two mothers. Eur J Obstet Gynecol Reprod Biol 1991, 38:233–238. CrossRef
- Romero R, Reece EA, Duff GW, Coultrip L, Hobbins JC: Prenatal diagnosis of Candida albicans chorioamnionitis. Am J Perinatol 1985, 2:121–122. CrossRef
- Bruner JP, Elliott JP, Kilbride HW, Garite TJ, Knox GE: Candida chorioamnionitis diagnosed by amniocentesis with subsequent fetal infection. Am J Perinatol 1986, 3:213–218. CrossRef
- Berry DL, Olson GL, Wen TS, Belfort MA, Moise KJ Jr: Candida chorioamnionitis: a report of two cases. J Matern Fetal Med 1997, 6:151–154. CrossRef
- Baley JE: Neonatal candidiasis: the current challenge. Clin Perinatol 1991, 18:263–280.
- Ng PC, Siu YK, Lewindon PJ, Wong W, Cheung KL, Dawkins R: Congenital Candida pneumonia in a preterm infant. J Paediatr Child Health 1994, 30:552–554. CrossRef
- Engelhart CM, van de Vijver NM, Nienhuis SJ, Hasaart TH: Fetal Candida sepsis at midgestation: a case report. Eur J Obstet Gynecol Reprod Biol 1998, 77:107–109. CrossRef
- Krishnakumari V, Rangaraj N, Nagaraj R: Antifungal activities of human beta-defensins HBD-1 to HBD-3 and their C-terminal analogs Phd1 to Phd3. Antimicrob Agents Chemother 2009, 53:256–260. CrossRef
- Feng Z, Jiang B, Chandra J, Ghannoum M, Nelson S, Weinberg A: Human beta-defensins: differential activity against candidal species and regulation by Candida albicans. J Dent Res 2005, 8:445–450.
- Lupetti A, Danesi R, Wout JW V't, Van Dissel JT, Senesi S, Nibbering PH: Antimicrobial peptides: therapeutic potential for the treatment of Candida infections. Expert Opin Investig Drugs 2002, 11:309–318. CrossRef
- Zaga V, Estrada-Gutierrez G, Beltran-Montoya J, Maida-Claros R, Lopez-Vancell R, Vadillo-Ortega F: Secretions of interleukin-1 beta and tumor necrosis factor alpha by whole fetal membranes depend on initial interactions of amnion or choriodecidua with lipopolysaccharides or group B streptococci. Biol Reprod 2004, 71:1296–1302. CrossRef
- Thiex NW, Chames MC, Loch-Caruso RK: Tissue-specific cytokine release from human extra-placental membranes stimulated by lipopolysaccharide in a two-compartment tissue culture system. Reprod Biol Endocrinol 2009, 7:117. CrossRef
- Miller MF, Loch-Caruso R: Comparison of LPS-stimulated release of cytokines in punch versus transwell tissue culture systems of human gestational membranes. Reprod Biol Endocrinol 2010, 8:121. CrossRef
- Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976, 72:248–254. CrossRef
- Zaga-Clavellina V, López GG, Estrada-Gutierrez G, Martinez-Flores A, Maida-Claros R, Beltran-Montoya J, Vadillo-Ortega F: Incubation of human chorioamniotic membranes with Candida albicans induces differential synthesis and secretion of interleukin-1beta, interleukin-6, prostaglandin E, and 92 kDa type IV collagenase. Mycoses 2006, 49:6–13. CrossRef
- Abrahams VM: Pattern recognition at the maternal-fetal interface. Immunol Invest 2008, 37:427–447. CrossRef
- Gow NA, Knox Y, Munro CA, Thompson WD: Infection of chick chorioallantoic membrane (CAM) as a model for invasive hyphal growth and pathogenesis of Candida albicans. Med Mycol 2003, 41:331–338. CrossRef
- Jacobsen ID, Grosse K, Berndt A, Hube B: Pathogenesis of Candida albicans infections in the alternative chorio-allantoic membrane chicken embryo model resembles systemic murine infections. PLoS One 2011, 6:e19741. CrossRef
- Velemínský M, Tosner J: Relationship of vaginal microflora to PROM, pPROM and the risk of early-onset neonatal sepsis. Neuro Endocrinol Lett 2008, 29:205–221.
- Frezza S, Maggio L, De Carolis MP, Gallini F, Puopolo M, Polimeni V, Costa S, Vento G, Tortorolo G: Risk factors for pulmonary candidiasis in preterm infants with a birth weight of less than 1250 g. Eur J Pediatr 2005, 164:88–92. CrossRef
- Gürgan T, Diker KS, Haziroglu R, Urman B, Akan M: In vitro infection of human fetal membranes with Candida species. Gynecol Obstet Invest 1994, 37:164–167. CrossRef
- Wiechuła B, Cholewa K, Ekiel A, Romanik M, Dolezych H, Martirosian G: HBD-1 and hBD-2 are expressed in cervico-vaginal lavage in female genital tract due to microbial infections. Ginekol Pol 2010, 81:268–271.
- Liu Z, Chen L: Congenital anti-candida of human vaginal epithelial cells. Beijing Da Xue Xue Bao. 2008, 40:174–177.
- Sørensen OE, Thapa DR, Rosenthal A, Liu L, Roberts AA, Ganz T: Differential regulation of beta-defensin expression in human skin by microbial stimuli. J Immunol 2005, 174:4870–4879.
- King AE, Fleming DC, Critchley HO, Kelly RW: Regulation of natural antibiotic expression by inflammatory mediators and mimics of infection in human endometrial epithelial cells. Mol Hum Reprod 2002, 8:341–349. CrossRef
- Krisanaprakornkit S, Kimball JR, Weinberg A, Darveau RP, Bainbridge BW, Dale BA: Inducible expression of human beta-defensin 2 by Fusobacterium nucleatum in oral epithelial cells: multiple signaling pathways and role of commensal bacteria in innate immunity and the epithelial barrier. Infect Immun 2000, 68:2907–2915. CrossRef
- Mathews M, Jia HP, Guthmiller JM, Losh G, Graham S, Johnson GK, Tack BF, McCray PB Jr: Production of beta-defensin antimicrobial peptides by the oral mucosa and salivary glands. Infect Immun 1999, 67:2740–2745.
- Harder J, Meyer-Hoffert U, Teran LM, Schwichtenberg L, Bartels J, Maune S, Schröder JM: 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.
- O'Neil DA, Porter EM, Elewaut D, Anderson GM, Eckmann L, Ganz T, Kagnoff MF: Expression and regulation of the human beta-defensins hBD-1 and hBD-2 in intestinal epithelium. J Immunol 1999, 163:6718–6724.
- Schröder JM, Harder J: Human beta-defensin-2. Int J Biochem Cell Biol 1999, 31:645–651. CrossRef
- Hiemstra PS: Epithelial antimicrobial peptides and proteins: their role in host defence and inflammation. Paediatr Respir Rev 2001, 2:306–310. CrossRef
- Vylkova S, Nayyar N, Li W, Edgerton M: Human beta-defensins kill Candida albicans in an energy-dependent and salt-sensitive manner without causing membrane disruption. Antimicrob Agents Chemother 2007, 51:154–161. CrossRef
- Harder J, Bartels J, Christophers E, Schroder JM: Isolation and characterization of human beta -defensin-3, a novel human inducible peptide antibiotic. J Biol Chem 2001, 276:5707–5713. CrossRef
- Steubesand N, Kiehne K, Brunke G, Pahl R, Reiss K, Herzig KH, Schubert S, Schreiber S, Fölsch UR, Rosenstiel P, Arlt A: The expression of the beta-defensins hBD-2 and hBD-3 is differentially regulated by NF-kappaB and MAPK/AP-1 pathways in an in vitro model of Candida esophagitis. BMC Immunol 2009, 10:36. CrossRef
- Joly S, Maze C, McCray PB Jr, Guthmiller JM: Human beta-defensins 2 and 3 demonstrate strain-selective activity against oral microorganisms. J Clin Microbiol 2004, 42:1024–1029. CrossRef
- Kim YM, Romero R, Chaiworapongsa T, Kim GJ, Kim MR, Kuivaniemi H, Tromp G, Espinoza J, Bujold E, Abrahams VM, Mor G: Toll-like receptor-2 and -4 in the chorioamniotic membranes in spontaneous labor at term and in preterm parturition that are associated with chorioamnionitis. Am J Obstet Gynecol 2004, 191:1346–1355. CrossRef
- Patni S, Flynn P, Wynen LP, Seager AL, Morgan G, White JO, Thornton CA: An introduction to Toll-like receptors and their possible role in the initiation of labour. BJOG 2007, 114:1326–1334. CrossRef
- Netea MG, Gow NA, Munro CA, Bates S, Collins C, Ferwerda G, Hobson RP, Bertram G, Hughes HB, Jansen T, Jacobs L, Buurman ET, Gijzen K, Williams DL, Torensma R, McKinnon A, MacCallum DM, Odds FC, Van der Meer JW, Brown AJ, Kullberg BJ: Immune sensing of Candida albicans requires cooperative recognition of mannans and glucans by lectin and Toll-like receptors. J Clin Invest 2006, 116:1642–1650. CrossRef
- Jouault T, El Abed-El Behi M, Martínez-Esparza M, Breuilh L, Trinel PA, Chamaillard M, Trottein F, Poulain D: Specific recognition of Candida albicans by macrophages requires galectin-3 to discriminate Saccharomyces cerevisiae and needs association with TLR2 for signaling. J Immunol 2006, 177:4679–4687.
- Bellocchio S, Montagnoli C, Bozza S, Gaziano R, Rossi G, Mambula SS, Vecchi A, Mantovani A, Levitz SM, Romani L: The contribution of the Toll-like/IL-1 receptor superfamily to innate and adaptive immunity to fungal pathogens in vivo. J Immunol 2004, 172:3059–3069.
- Tissue-specific human beta-defensins (HBD)-1, HBD-2 and HBD-3 secretion profile from human amniochorionic membranes stimulated with Candida albicans in a two-compartment tissue culture system
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Reproductive Biology and Endocrinology
- Online Date
- September 2012
- Online ISSN
- BioMed Central
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- Author Affiliations
- 1. Department of Cell Biology, Instituto Nacional de Perinatologia “Isidro Espinosa de los Reyes”, Mexico City, Mexico
- 2. Department of Nutrition Research, Instituto Nacional de Perinatologia “Isidro Espinosa de los Reyes”, Mexico City, Mexico
- 3. Department of Biochemistry and Molecular Biology, Instituto Nacional de Perinatologia “Isidro Espinosa de los Reyes”, Mexico City, Mexico
- 4. Department of Infectology, Instituto Nacional de Perinatologia “Isidro Espinosa de los Reyes”, Mexico City, Mexico
- 5. Department of Family Planning, Department of Gynecology and Obstetrics, Instituto Nacional de Perinatologia “Isidro Espinosa de los Reyes”, Mexico City, Mexico