Top

Reproductive Biology and Endocrinology

Published in:

Open Access 01-12-2010 | Research

Comparison of LPS-stimulated release of cytokines in punch versus transwell tissue culture systems of human gestational membranes

Authors: Mark F Miller, Rita Loch-Caruso

Published in: Reproductive Biology and Endocrinology | Issue 1/2010

Abstract

Background

Cytokine signaling within the amnionic, chorionic and decidual extraplacental gestational membranes plays an important role in membrane rupture and the timing of birth. The predominant in vitro explant culture system for evaluating cytokine induction in human gestational membranes has been the free-floating biopsy punch culture. Punch systems have been used to investigate the impact of various toxicants, pharmaceuticals and genetic variation on expression of pro-inflammatory cytokines. More recently, a dual compartment transwell culture system has been developed that more closely mimics the intrauterine compartment. The current study compares these two systems with respect to release of pro- and anti-inflammatory cytokines in response to lipopolysaccharide (LPS), a model stimulant.

Methods

Tissue samples were exposed to 100 ng/ml LPS for 12 h and cytokines were measured by ELISA. Data are expressed as increase relative to non-treated controls.

Results

Levels of interleukin-6 increased in punch culture medium samples to a significantly greater extent (34.2 fold) compared with medium from transwell cultures in the amnion (6.6 fold) or choriodecidual (7.1 fold) compartments. Interleukin-8 also showed a significantly greater induction in punch (4.8 fold) than transwell amnion (1.6 fold) or choriodecidual (1.7 fold) samples. The anti-inflammatory interleukin-10 showed a significant difference between punch (36.5 fold) and transwell amnion (15.4 fold) samples, but no difference was observed between punch and transwell choriodecidual (28.5 fold) samples. Neither interleukin-1beta nor tumor necrosis factor-alpha (TNF-alpha) showed a significant difference between the punch and transwell samples.

Conclusions

These results indicate that the pattern of LPS-stimulated cytokine release from gestational membranes in vitro depends on the culture system used, confounding comparisons of studies that use different gestational membrane culture systems to study inflammatory responses.

Available only for authorised users

Hung TH, Chen SF, Hsu JJ, Hsieh CC, Hsueh S, Hsieh TT: Tumour necrosis factor-alpha converting enzyme in human gestational tissues from pregnancies complicated by chorioamnionitis. Placenta. 2006, 27 (9-10): 996-1006. 10.1016/j.placenta.2005.11.002.CrossRefPubMed

Menon R, Merialdi M, Lombardi SJ, Fortunato SJ: Differences in the placental membrane cytokine response: a possible explanation for the racial disparity in preterm birth. Am J Reprod Immunol. 2006, 56 (2): 112-118. 10.1111/j.1600-0897.2006.00394.x.CrossRefPubMed

Han YM, Romero R, Kim JS, Tarca AL, Kim SK, Draghici S, Kusanovic JP, Gotsch F, Mittal P, Hassan SS, Kim CJ: Region-specific gene expression profiling: novel evidence for biological heterogeneity of the human amnion. Biol Reprod. 2008, 79 (5): 954-961. 10.1095/biolreprod.108.069260.PubMedCentralCrossRefPubMed

Simpson KL, Keelan JA, Mitchell MD: Labour-associated changes in the regulation of production of immunomodulators in human amnion by glucocorticoids, bacterial lipopolysaccharide and pro-inflammatory cytokines. J Reprod Fertil. 1999, 116 (2): 321-327. 10.1530/jrf.0.1160321.CrossRefPubMed

Aaltonen R, Heikkinen J, Vahlberg T, Jensen JS, Alanen A: Local inflammatory response in choriodecidua induced by Ureaplasma urealyticum. BJOG. 2007, 114 (11): 1432-1435. 10.1111/j.1471-0528.2007.01410.x.CrossRefPubMed

Mitchell MD, Sato TA, Wang A, Keelan JA, Ponnampalam AP, Glass M: Cannabinoids stimulate prostaglandin production by human gestational tissues through a tissue- and CB1-receptor-specific mechanism. Am J Physiol Endocrinol Metab. 2008, 294 (2): E352-356. 10.1152/ajpendo.00495.2007.CrossRefPubMed

Fortunato SJ, Menon RP, Swan KF, Menon R: Inflammatory cytokine (interleukins 1, 6 and 8 and tumor necrosis factor-alpha) release from cultured human fetal membranes in response to endotoxic lipopolysaccharide mirrors amniotic fluid concentrations. Am J Obstet Gynecol. 1996, 174 (6): 1855-1861. 10.1016/S0002-9378(96)70221-1. discussion 1861-1862CrossRefPubMed

Lappas M, Permezel M, Rice GE: 15-Deoxy-delta(12,14)-prostaglandin J(2) and troglitazone regulation of the release of phospholipid metabolites, inflammatory cytokines and proteases from human gestational tissues. Placenta. 2006, 27 (11-12): 1060-1072. 10.1016/j.placenta.2005.11.009.CrossRefPubMed

Sato TA, Keelan JA, Mitchell MD: Critical paracrine interactions between TNF-alpha and IL-10 regulate lipopolysaccharide-stimulated human choriodecidual cytokine and prostaglandin E2 production. J Immunol. 2003, 170 (1): 158-166.CrossRefPubMed

10.

Simpson KL, Keelan JA, Mitchell MD: Labor-associated changes in interleukin-10 production and its regulation by immunomodulators in human choriodecidua. J Clin Endocrinol Metab. 1998, 83 (12): 4332-4337. 10.1210/jc.83.12.4332.CrossRefPubMed

11.

Helliwell RJ, Keelan JA, Marvin KW, Adams L, Chang MC, Anand A, Sato TA, O'Carroll S, Chaiworapongsa T, Romero RJ, Mitchell MD: Gestational age-dependent up-regulation of prostaglandin D synthase (PGDS) and production of PGDS-derived antiinflammatory prostaglandins in human placenta. J Clin Endocrinol Metab. 2006, 91 (2): 597-606. 10.1210/jc.2005-1982.CrossRefPubMed

12.

Lappas M, Yee K, Permezel M, Rice GE: Release and regulation of leptin, resistin and adiponectin from human placenta, fetal membranes, and maternal adipose tissue and skeletal muscle from normal and gestational diabetes mellitus-complicated pregnancies. J Endocrinol. 2005, 186 (3): 457-465. 10.1677/joe.1.06227.CrossRefPubMed

13.

Zaga V, Estrada-Gutierrez G, Beltran-Montoya J, Maida-Claros R, Lopez-Vancell R, Vadillo-Ortega F: Secretions of interleukin-1beta 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 (4): 1296-1302. 10.1095/biolreprod.104.028621.CrossRefPubMed

14.

Zaga-Clavellina V, Garcia-Lopez G, Flores-Herrera H, Espejel-Nunez A, Flores-Pliego A, Soriano-Becerril D, Maida-Claros R, Merchant-Larios H, Vadillo-Ortega F: In vitro secretion profiles of interleukin (IL)-1beta, IL-6, IL-8, IL-10, and TNF alpha after selective infection with Escherichia coli in human fetal membranes. Reprod Biol Endocrinol. 2007, 5 (1): 46-10.1186/1477-7827-5-46.PubMedCentralCrossRefPubMed

15.

Zaga-Clavellina V, Lopez 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 (1): 6-13. 10.1111/j.1439-0507.2005.01171.x.CrossRefPubMed

16.

Thiex N, Chames M, Loch-Caruso R: Tissue-specific cytokine release from human extra-placental membranes stimulated by lipopolysaccharide in a two-compartment tissue culture system. Reprod Biol Endocrinol. 2009, 7 (1): 117-10.1186/1477-7827-7-117.PubMedCentralCrossRefPubMed

17.

Keelan JA, Khan S, Yosaatmadja F, Mitchell MD: Prevention of inflammatory activation of human gestational membranes in an ex vivo model using a pharmacological NF-{kappa}B inhibitor. J Immunol. 2009, 183 (8): 5270-5278. 10.4049/jimmunol.0802660.CrossRefPubMed

18.

Al-Asmakh M, Race H, Tan S, Sullivan MH: The effects of oxygen concentration on in vitro output of prostaglandin E2 and interleukin-6 from human fetal membranes. Mol Hum Reprod. 2007, 13 (3): 197-201. 10.1093/molehr/gal109.CrossRefPubMed

19.

Funai EF, Labowsky AT, Drewes CE, Kliman HJ: Timing of fetal meconium absorption by amnionic macrophages. Am J Perinatol. 2009, 26 (1): 93-97. 10.1055/s-0028-1103028.CrossRefPubMed

20.

Kent AS, Sullivan MH, Elder MG: Transfer of cytokines through human fetal membranes. J Reprod Fertil. 1994, 100 (1): 81-84. 10.1530/jrf.0.1000081.CrossRefPubMed

21.

Kjaergaard N, Helmig RB, Schonheyder HC, Uldbjerg N, Hansen ES, Madsen H: Chorioamniotic membranes constitute a competent barrier to group b streptococcus in vitro. Eur J Obstet Gynecol Reprod Biol. 1999, 83 (2): 165-169. 10.1016/S0301-2115(99)00009-3.CrossRefPubMed

22.

Rajasingam D, Bennett PR, Alvi SA, Elder MG, Sullivan MH: Stimulation of prostaglandin production from intact human fetal membranes by bacteria and bacterial products. Placenta. 1998, 19 (4): 301-306. 10.1016/S0143-4004(98)90062-5.CrossRefPubMed

23.

Bethin KE, Nagai Y, Sladek R, Asada M, Sadovsky Y, Hudson TJ, Muglia LJ: Microarray analysis of uterine gene expression in mouse and human pregnancy. Mol Endocrinol. 2003, 17 (8): 1454-1469. 10.1210/me.2003-0007.CrossRefPubMed

24.

Marvin KW, Keelan JA, Eykholt RL, Sato TA, Mitchell MD: Use of cDNA arrays to generate differential expression profiles for inflammatory genes in human gestational membranes delivered at term and preterm. Mol Hum Reprod. 2002, 8 (4): 399-408. 10.1093/molehr/8.4.399.CrossRefPubMed

25.

Gibbs RS, Romero R, Hillier SL, Eschenbach DA, Sweet RL: A review of premature birth and subclinical infection. Am J Obstet Gynecol. 1992, 166 (5): 1515-1528.CrossRefPubMed

26.

Bowen JM, Chamley L, Keelan JA, Mitchell MD: Cytokines of the placenta and extra-placental membranes: roles and regulation during human pregnancy and parturition. Placenta. 2002, 23 (4): 257-273. 10.1053/plac.2001.0782.CrossRefPubMed

27.

Karim SM, Trussell RR, Patel RC, Hillier K: Response of pregnant human uterus to prostaglandin-F2-alpha-induction of labour. Br Med J. 1968, 4 (631): 621-623. 10.1136/bmj.4.5631.621.PubMedCentralCrossRefPubMed

28.

Challis JRG, Lye SJ: Parturition. The Physiology of Reproduction. Edited by: Knobil E, Neill JD. 1994, New York: Raven Press, 985-1013.

29.

Keirse MJNC: Eicosanoids in human pregnancy and parturition. Eicosanoids and Reproduction. Edited by: Mitchell MD. 1990, Boca Raton: Press Ltd

30.

Opsjln SL, Wathen NC, Tingulstad S, Wiedswang G, Sundan A, Waage A, Austgulen R: Tumor necrosis factor, interleukin-1, and interleukin-6 in normal human pregnancy. Am J Obstet Gynecol. 1993, 169 (2Pt 1): 397-404.CrossRefPubMed

31.

Tsuda A, Ikegami T, Hirano H, Sanada H, Ogawa M, Sasaki M, Tanaka T: The relationship between amniotic fluid interleukin-6 concentration and histologic evidence of chorioamnionitis. Acta Obstet Gynecol Scand. 1998, 77 (5): 515-520. 10.1080/j.1600-0412.1998.770508.x.CrossRefPubMed

32.

Romero R, Avila C, Santhanam U, Sehgal PB: Amniotic fluid interleukin 6 in preterm labor. Association with infection. J Clin Invest. 1990, 85 (5): 1392-1400. 10.1172/JCI114583.PubMedCentralCrossRefPubMed

33.

Santhanam U, Avila C, Romero R, Viguet H, Ida N, Sakurai S, Sehgal PB: Cytokines in normal and abnormal parturition: elevated amniotic fluid interleukin-6 levels in women with premature rupture of membranes associated with intrauterine infection. Cytokine. 1991, 3 (2): 155-163. 10.1016/1043-4666(91)90037-E.CrossRefPubMed

34.

Jauniaux E, Gulbis B, Schandene L, Collette J, Hustin J: Distribution of interleukin-6 in maternal and embryonic tissues during the first trimester. Mol Hum Reprod. 1996, 2 (4): 239-243. 10.1093/molehr/2.4.239.CrossRefPubMed

35.

Stallmach T, Hebisch G, Joller-Jemelka HI, Orban P, Schwaller J, Engelmann M: Cytokine production and visualized effects in the feto-maternal unit. Quantitative and topographic data on cytokines during intrauterine disease. Lab Invest. 1995, 73 (3): 384-392.PubMed

36.

Menon R, Fortunato SJ: Fetal membrane inflammatory cytokines: a switching mechanism between the preterm premature rupture of the membranes and preterm labor pathways. J Perinat Med. 2004, 32 (5): 391-399. 10.1515/JPM.2004.134.CrossRefPubMed

37.

Moore KW, O'Garra A, de Waal Malefyt R, Vieira P, Mosmann TR: Interleukin-10. Annu Rev Immunol. 1993, 11: 165-190. 10.1146/annurev.iy.11.040193.001121.CrossRefPubMed

38.

Daftarian PM, Kumar A, Kryworuchko M, Diaz-Mitoma F: IL-10 production is enhanced in human T cells by IL-12 and IL-6 and in monocytes by tumor necrosis factor-alpha. J Immunol. 1996, 157 (1): 12-20.PubMed

39.

Hashii K, Fujiwara H, Yoshioka S, Kataoka N, Yamada S, Hirano T, Mori T, Fujii S, Maeda M: Peripheral blood mononuclear cells stimulate progesterone production by luteal cells derived from pregnant and non-pregnant women: possible involvement of interleukin-4 and interleukin-10 in corpus luteum function and differentiation. Hum Reprod. 1998, 13 (1O): 2738-2744.CrossRefPubMed

40.

Robertson SA, Skinner RJ, Care AS: Essential role for IL-10 in resistance to lipopolysaccharide-induced preterm labor in mice. J Immunol. 2006, 177 (7): 4888-4896.CrossRefPubMed

41.

Tewari N, Kalkunte S, Murray DW, Sharma S: The water channel aquaporin 1 is a novel molecular target of polychlorinated biphenyls for in utero anomalies. J Biol Chem. 2009, 284 (22): 15224-15232. 10.1074/jbc.M808892200.PubMedCentralCrossRefPubMed

42.

Lappas M, Rice GE: The role and regulation of the nuclear factor kappa B signalling pathway in human labour. Placenta. 2007, 28 (5-6): 543-556. 10.1016/j.placenta.2006.05.011.CrossRefPubMed

Title: Comparison of LPS-stimulated release of cytokines in punch versus transwell tissue culture systems of human gestational membranes
Authors: Mark F Miller
Rita Loch-Caruso
Publication date: 01-12-2010
Publisher: BioMed Central
Published in: Reproductive Biology and Endocrinology / Issue 1/2010
Electronic ISSN: 1477-7827
DOI: https://doi.org/10.1186/1477-7827-8-121

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Comparison of LPS-stimulated release of cytokines in punch versus transwell tissue culture systems of human gestational membranes

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2010

Ovarian matrix metalloproteinases are differentially regulated during the estrous cycle but not during short photoperiod induced regression in Siberian hamsters (Phodopus sungorus)

TRIzol treatment of secretory phase endometrium allows combined proteomic and mRNA microarray analysis of the same sample in women with and without endometriosis

Effects of luteectomy in early pregnancy on the maintenance of gestation and plasma progesterone concentrations in the viviparous temperate lizard Barisia imbricata imbricata

Metabolic abnormalities in adolescents with polycystic ovary syndrome in south china

Can egg donor selection be improved? - a pilot study

Changes in cell proliferation, but not in vascularisation are characteristic for human endometrium in different reproductive failures - a pilot study