Group B Streptococcal Infection of the Choriodecidua Induces Dysfunction of the Cytokeratin Network in Amniotic Epithelium: A Pathway to Membrane Weakening

Early events leading to intrauterine infection remain poorly defined, but may hold the key to preventing preterm delivery. To determine molecular pathways within fetal membranes (chorioamnion) associated with early choriodecidual infection that may progress to preterm premature rupture of membranes (PPROM), we examined the effects of a Group B Streptococcus (GBS) choriodecidual infection on chorioamnion in a nonhuman primate model. Ten chronically catheterized pregnant monkeys (Macaca nemestrina) at 118–125 days gestation (term = 172 days) received choriodecidual inoculation of either GBS (n = 5) or saline (n = 5). Cesarean section was performed in the first week after GBS or saline inoculation. RNA extracted from chorioamnion (inoculation site) was profiled by microarray. Single gene, Gene Set, and Ingenuity Pathway Analysis results were validated using qRT-PCR (chorioamnion), Luminex (amniotic fluid, AF), immunohistochemistry, and transmission electron microscopy (TEM). Despite uterine quiescence in most cases, significant elevations of AF cytokines (TNF-α, IL-8, IL-1β, IL-6) were detected in GBS versus controls (p<0.05). Choriodecidual infection resolved by the time of cesarean section in 3 of 5 cases and GBS was undetectable by culture and PCR in the AF. A total of 331 genes were differentially expressed (>2-fold change, p<0.05). Remarkably, GBS exposure was associated with significantly downregulated expression of multiple cytokeratin (CK) and other cytoskeletal genes critical for maintenance of tissue tensile strength. Immunofluorescence revealed highly significant changes in the CK network within amniocytes with dense CK aggregates and retraction from the cell periphery (all p = 0.006). In human pregnancies affected by PPROM, there was further evidence of CK network retraction with significantly shorter amniocyte foot processes (p = 0.002). These results suggest early choriodecidual infection results in decreased cellular membrane integrity and tensile strength via dysfunction of CK networks. Downregulation of CK expression and perturbations in the amniotic epithelial cell intermediate filament network occur after GBS choriodecidual infection, which may contribute to PPROM.

[1]  L. Aravind,et al.  A hemolytic pigment of Group B Streptococcus allows bacterial penetration of human placenta , 2013, The Journal of experimental medicine.

[2]  R. Bentley,et al.  The Chorion Layer of Fetal Membranes Is Prematurely Destroyed in Women With Preterm Premature Rupture of the Membranes , 2013, Reproductive Sciences.

[3]  D. Tian,et al.  Correlation between apoptosis and TGF-β1 expression in the mucosal epithelium of rat small intestine in a cold stress state , 2013, Experimental and therapeutic medicine.

[4]  J. Strauss.  Extracellular Matrix Dynamics and Fetal Membrane Rupture , 2013, Reproductive Sciences.

[5]  H. D. Liggitt,et al.  Choriodecidual Infection Downregulates Angiogenesis and Morphogenesis Pathways in Fetal Lungs from Macaca Nemestrina , 2012, PloS one.

[6]  A. Mes-Masson,et al.  p53 inhibits angiogenesis by inducing the production of Arresten. , 2012, Cancer research.

[7]  S. Dramsi,et al.  Group B Streptococcus GAPDH Is Released upon Cell Lysis, Associates with Bacterial Surface, and Induces Apoptosis in Murine Macrophages , 2012, PloS one.

[8]  H. D. Liggitt,et al.  Choriodecidual Group B Streptococcal Inoculation Induces Fetal Lung Injury without Intra-Amniotic Infection and Preterm Labor in Macaca nemestrina , 2011, PloS one.

[9]  M. Beil,et al.  Cytoskeleton in motion: the dynamics of keratin intermediate filaments in epithelia , 2011, The Journal of cell biology.

[10]  N. Gomez‐Lopez,et al.  Specific inflammatory microenvironments in the zones of the fetal membranes at term delivery. , 2011, American journal of obstetrics and gynecology.

[11]  B. Mercer,et al.  Preterm PROM: prediction, prevention, principles. , 2011, Clinical obstetrics and gynecology.

[12]  S. Fortunato,et al.  Cigarette smoke induces oxidative stress and apoptosis in normal term fetal membranes. , 2011, Placenta.

[13]  Chengyu Liang,et al.  Evidence that inhibition of BAX activation by BCL-2 involves its tight and preferential interaction with the BH3 domain of BAX , 2011, Cell Research.

[14]  M. Gravett,et al.  Use of nonhuman primate models to investigate mechanisms of infection‐associated preterm birth , 2011, BJOG : an international journal of obstetrics and gynaecology.

[15]  S. Dramsi,et al.  The surface protein HvgA mediates group B streptococcus hypervirulence and meningeal tropism in neonates , 2010, The Journal of experimental medicine.

[16]  M. Gravett,et al.  Differential host response to LPS variants in amniochorion and the TLR4/MD-2 system in Macaca nemestrina. , 2010, Placenta.

[17]  T. Aach,et al.  The keratin-filament cycle of assembly and disassembly , 2010, Journal of Cell Science.

[18]  H. Bradshaw,et al.  Uterine-specific p53 deficiency confers premature uterine senescence and promotes preterm birth in mice. , 2010, The Journal of clinical investigation.

[19]  Y. Matsuzaki,et al.  Overview of epidermolysis bullosa , 2010, The Journal of dermatology.

[20]  K. Ridge,et al.  The role of the ubiquitin proteasome pathway in keratin intermediate filament protein degradation. , 2010, Proceedings of the American Thoracic Society.

[21]  R. Menon,et al.  Diversity in cytokine response to bacteria associated with preterm birth by fetal membranes. , 2009, American journal of obstetrics and gynecology.

[22]  Jörg Rahnenführer,et al.  Robert Gentleman, Vincent Carey, Wolfgang Huber, Rafael Irizarry, Sandrine Dudoit (2005): Bioinformatics and Computational Biology Solutions Using R and Bioconductor , 2009 .

[23]  Z-Y Shen,et al.  Autophagic and apoptotic cell death in amniotic epithelial cells. , 2008, Placenta.

[24]  Daniel B. DiGiulio,et al.  Microbial Prevalence, Diversity and Abundance in Amniotic Fluid During Preterm Labor: A Molecular and Culture-Based Investigation , 2008, PloS one.

[25]  C. Ockleford,et al.  Immunofluorescence confocal laser scanning microscopy and immuno-electron microscopic identification of keratins in human materno-foetal interaction zone , 2008, Journal of cellular and molecular medicine.

[26]  J. Sever,et al.  Group B Streptococcus and pregnancy: a review. , 2008, American journal of obstetrics and gynecology.

[27]  D. Persing,et al.  Pretreatment With Toll-like Receptor 4 Antagonist Inhibits Lipopolysaccharide-Induced Preterm Uterine Contractility, Cytokines, and Prostaglandins in Rhesus Monkeys , 2008, Reproductive Sciences.

[28]  Roberto Romero,et al.  Epidemiology and causes of preterm birth , 2008, The Lancet.

[29]  G. Buonocore,et al.  Association between oxidative stress in pregnancy and preterm premature rupture of membranes. , 2007, Clinical biochemistry.

[30]  R. Tibshirani,et al.  On testing the significance of sets of genes , 2006, math/0610667.

[31]  M. Oyen,et al.  Premature rupture of the fetal membranes: is the amnion the major determinant? , 2006, American journal of obstetrics and gynecology.

[32]  R. Mannucci,et al.  Group B Streptococcus Induces Macrophage Apoptosis by Calpain Activation1 , 2006, The Journal of Immunology.

[33]  Robert M. Moore,et al.  Fetal Membranes From Term VAginal Deliveries Have a Zone of Weakness Exhibiting Characteristics of Apoptosis and Remodeling , 2006, The Journal of the Society for Gynecologic Investigation: JSGI.

[34]  W. Kuri-Harcuch,et al.  Interleukin-6 promotes human epidermal keratinocyte proliferation and keratin cytoskeleton reorganization in culture , 2006, Cell and Tissue Research.

[35]  R. Rappuoli,et al.  Group B Streptococcus crosses human epithelial cells by a paracellular route. , 2006, The Journal of infectious diseases.

[36]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[37]  J. Cleveland,et al.  Mechanisms of Group B Streptococcal-Induced Apoptosis of Murine Macrophages 1 , 2005, The Journal of Immunology.

[38]  S. Artan,et al.  Programmed cell death (apoptosis) in placentas from normal pregnancy and pregnancy complicated by term (t) and preterm (p) premature rupture of membranes (PROM) , 2005, Archives of Gynecology and Obstetrics.

[39]  G. Ulett,et al.  Nitric oxide is a key determinant of group B streptococcus-induced murine macrophage apoptosis. , 2005, The Journal of infectious diseases.

[40]  M. Elovitz,et al.  Animal models of preterm birth , 2004, Trends in Endocrinology & Metabolism.

[41]  R. Menon,et al.  The Role of Matrix Degrading Enzymes and Apoptosis in Repture of Membranes , 2004, The Journal of the Society for Gynecologic Investigation: JSGI.

[42]  Jean YH Yang,et al.  Bioconductor: open software development for computational biology and bioinformatics , 2004, Genome Biology.

[43]  D. Bullard,et al.  Regulation of endothelial glutathione by ICAM‐1: implications for inflammation , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[44]  Rafael A. Irizarry,et al.  A Model-Based Background Adjustment for Oligonucleotide Expression Arrays , 2004 .

[45]  R. Windoffer,et al.  Identification of novel principles of keratin filament network turnover in living cells. , 2004, Molecular biology of the cell.

[46]  J. Potts,et al.  The molecular basis of fibronectin‐mediated bacterial adherence to host cells , 2004, Molecular microbiology.

[47]  Gordon K Smyth,et al.  Statistical Applications in Genetics and Molecular Biology Linear Models and Empirical Bayes Methods for Assessing Differential Expression in Microarray Experiments , 2011 .

[48]  E. Schuetz,et al.  Co-regulation of CYP3A4 and CYP3A5 and contribution to hepatic and intestinal midazolam metabolism. , 2002, Molecular pharmacology.

[49]  R. Oshima,et al.  Apoptosis and keratin intermediate filaments , 2002, Cell Death and Differentiation.

[50]  I. Furuta,et al.  Increased apoptosis of human fetal membranes in rupture of the membranes and chorioamnionitis. , 2002, Placenta.

[51]  T. Kavanagh,et al.  Tissue specific changes in the expression of glutamate-cysteine ligase mRNAs in mice exposed to methylmercury. , 2001, Toxicology letters.

[52]  J. Woods.  Reactive oxygen species and preterm premature rupture of membranes-a review. , 2001, Placenta.

[53]  V. Nizet,et al.  Genetic basis for the β‐haemolytic/cytolytic activity of group B Streptococcus , 2001, Molecular microbiology.

[54]  S. Fortunato,et al.  Programmed cell death (apoptosis) as a possible pathway to metalloproteinase activation and fetal membrane degradation in premature rupture of membranes. , 2000, American journal of obstetrics and gynecology.

[55]  J. Hauth,et al.  Intrauterine infection and preterm delivery. , 2000, The New England journal of medicine.

[56]  M. Omary,et al.  Keratins Turn over by Ubiquitination in a Phosphorylation-Modulated Fashion , 2000, The Journal of cell biology.

[57]  G. Tamura,et al.  Group B Streptococci and Other Gram-Positive Cocci Bind to Cytokeratin 8 , 2000, Infection and Immunity.

[58]  R. Flavell,et al.  Abrogation of TGFβ Signaling in T Cells Leads to Spontaneous T Cell Differentiation and Autoimmune Disease , 2000 .

[59]  D. Lawrence,et al.  Growth stimulation of murine fibroblasts by TGF-β1 depends on the expression of a functional p53 protein , 1999, Oncogene.

[60]  S. Haldar,et al.  The relationship between BcI2, Bax and p53: consequences for cell cycle progression and cell death. , 1998, Molecular human reproduction.

[61]  A. Roberts,et al.  Autocrine secretion of TGF-β1 and TGF-β2 by pre-adipocytes and adipocytes: A potent negative regulator of adipocyte differentiation and proliferation of mammary carcinoma cells , 1998, In Vitro Cellular & Developmental Biology - Animal.

[62]  M. Krohn,et al.  Broad-spectrum bacterial rDNA polymerase chain reaction assay for detecting amniotic fluid infection among women in premature labor. , 1997, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[63]  B. Mercer,et al.  PRETERM LABOR AND PRETERM PREMATURE RUPTURE OF THE MEMBRANES Diagnosis and Management , 1997 .

[64]  E. Furth,et al.  A program of cell death and extracellular matrix degradation is activated in the amnion before the onset of labor. , 1996, The Journal of clinical investigation.

[65]  S. Witkin,et al.  An experimental model for intraamniotic infection and preterm labor in rhesus monkeys. , 1994, American journal of obstetrics and gynecology.

[66]  S. Bell,et al.  Structural characteristics of term human fetal membranes: a novel zone of extreme morphological alteration within the rupture site , 1994, British journal of obstetrics and gynaecology.

[67]  J. Fisher,et al.  Role of oxygen free radicals in cocaine-induced vascular disruption in mice. , 1994, Teratology.

[68]  R. Romero,et al.  The diagnostic and prognostic value of amniotic fluid white blood cell count, glucose, interleukin-6, and gram stain in patients with preterm labor and intact membranes. , 1993, American journal of obstetrics and gynecology.

[69]  N. Kiviat,et al.  The Relationship of Amniotic Fluid Cytokines and Preterm Delivery, Amniotic Fluid Infection, Histologic Chorioamnionitis, and Chorioamnion Infection , 1993, Obstetrics and gynecology.

[70]  E. Lieberman,et al.  Cigarettes, coffee, and preterm premature rupture of the membranes. , 1993, American journal of epidemiology.

[71]  R. Romero,et al.  Neutrophil attractant/activating peptide-1/interleukin-8 in term and preterm parturition. , 1991, American journal of obstetrics and gynecology.

[72]  J. Musser,et al.  Identification of a high-virulence clone of type III Streptococcus agalactiae (group B Streptococcus) causing invasive neonatal disease. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[73]  H. Denk,et al.  The distribution of intermediate filament proteins, actin and desmoplakins in human placental tissue as revealed by polyclonal and monoclonal antibodies , 1989, Placenta.

[74]  O. Itelson,et al.  [Premature rupture of fetal membranes]. , 1969, Harefuah.

[75]  Robert M. Moore,et al.  Proinflammatory Cytokines Found in Amniotic Fluid Induce Collagen Remodeling, Apoptosis, and Biophysical Weakening of Cultured Human Fetal Membranes1 , 2006, Biology of reproduction.

[76]  Emily Dimmer,et al.  The Gene Ontology Annotation (GOA) Database: sharing knowledge in Uniprot with Gene Ontology , 2004, Nucleic Acids Res..

[77]  C. Hernández-Guerrero,et al.  Identification of matrix metalloproteinase-9 in amniotic fluid and amniochorion in spontaneous labor and after experimental intrauterine infection or interleukin-1 beta infusion in pregnant rhesus monkeys. , 2002, American journal of obstetrics and gynecology.

[78]  R. Flavell,et al.  Abrogation of TGFbeta signaling in T cells leads to spontaneous T cell differentiation and autoimmune disease. , 2000, Immunity.

[79]  T. Mukhopadhyay,et al.  p53 involvement in activation of the cytokeratin 8 gene in tumor cell lines. , 1996, Anticancer research.

[80]  T. Martin,et al.  Lung antibacterial defense mechanisms in infant and adult rats: implications for the pathogenesis of group B streptococcal infections in the neonatal lung. , 1988, The Journal of infectious diseases.