56: Diversity in cytokine signature exhibited by bacterial pathogens associated with spontaneous preterm birth in human fetal membranes

SMFM Abstracts

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TIME-DEPENDENT EFFECT OF IN UTERO INFLAMMATION ON THE OFFSPRING’S BEHAVIOR: A LONGITUDINAL STUDY IN RATS MARIE-ÈVR ROY-LACROIX1, MAXIME GUÉRARD2, SYLVIE GIRARD3, MARYSE BERTHIAUME4, MAREK ROLA-PLESZCZYNSKI3, JEAN-CHARLES PASQUIER1, 1Centre Hospitalier de l’Université de Sherbrooke, Obstétrique Gynécologie, Sherbrooke, Quebec, Canada, 2Université de Sherbrooke, Biologie, Sherbooke, Quebec, Canada, 3Centre Hospitalier de l’Université de Sherbrooke, Pédiatrie, Sherbrooke, Quebec, Canada, 4Centre Hospitalier de l’Université de Sherbrooke, Centre de recherche clinique, Sherbrooke, Quebec, Canada OBJECTIVE: The foetal inflammatory response syndrome (FIRS) is a risk factor for the development of long-term sequelae but the correlation between the duration of in utero exposure to inflammation and the occurrence of neurologic damage is not well defined. The objective of the present study was to examine the effects of prenatal exposure to inflammation on motor performance of pups. STUDY DESIGN: Time-pregnant Sprague-Dawley rats (n⫽32) received intraperitoneal injection of LPS (500 ␮g/kg), or an equivalent volume of vehicle 3h, 6h,12h, and 24h before C-section performed at gestational age 22 days. Maternal serum and amniotic fluid were collected and tested for 19 cytokines. Four pups per rat were resuscitated and given to surrogate mothers. Motor activity of pups (n⫽58) were analyzed using the open field test at postnatal day 20. Brains of pups were collected at postnatal day 20 for histopathological examination. Analysis included ANOVA, Kaplan Meier and Student=s t-test with p ⬍0.05 considered significant. RESULTS: Perinatal mortality was significantly increased in the LPS group (48/ 84, 57%) compared with the control group (2/24, 8%) and mortality was significantly higher for groups exposed 6, 12 and 24h to LPS compared to the control group. Behavioural experiments showed that in utero exposure to inflammation decreased locomotor (p⬍0.01) and exploratory activity (p⫽0.06) of pups during a 5-min session in the open field test. Motor performances of pups exposed for 3 or 6 h to LPS were not significantly different from control pups but offspring exposed 12 h to LPS were weaker than control pups in term of distance (p⫽0.01) and speed (p⫽0.02) of locomotion. CONCLUSION: Our results showed that increasing time of intrauterine exposure to maternal inflammation induces, in their offspring, long-lasting neurobehavioral abnormalities. In our murine model, a threshold time of exposition to inflammation is clearly demonstrated, before which we should act to prevent permanent brain damage.

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REGULATION OF STEROID SULFATE TRANSPORTERS IN HUMAN PLACENTA DURING VARIOUS PREGNANCY RELATED DISEASE STATES AND THEIR IMPACT ON ESTROGEN SYNTHESIS PATHWAY CLIFFORD MASON1, CATALIN BUHIMSCHI2, IRINA BUHIMSCHI2, PETER SWAAN3, CARL P WEINER4, 1University of Maryland at Baltimore, Pharmaceutical Sciences, Baltimore, Maryland, 2Yale University, New Haven, Connecticut, 3 University of Maryland at Baltimore, Baltimore, Maryland, 4University of Kansas, Kansas City, Kansas OBJECTIVE: The critical uptake of steroid sulfates by syncytiotrophoblasts is predominately mediated by OAT4 and OATP-B transporters. However, it is unknown whether pregnancy related diseases alter transporter expression. We investigated the impact of pregnancy related disease states on the regulation of several steroid sulfate transporters and enzymes. STUDY DESIGN: Placenta were obtained from 5 groups of women at primary cesarean delivery (i) term no labor (n⫽10, TNL, 39.2 weeks); (ii) term labor (n⫽8, TL, 39.3 weeks); (iii) preterm no labor delivered for severe preeclampsia (n⫽10, PT-SPE, 30.3 weeks); (iv) preterm labor unassociated with inflammation (n⫽10, PTL-NI, 30.5 weeks); (v) preterm labor with inflammation (n⫽10, PTL-I, 28.7 weeks). Gene expression was determined by PCR and quantitative RT PCR. Western blot analyses were performed to quantify changes in estrogen receptor isoforms. RESULTS: While OAT4 and OATP-B were expressed in all placental samples, only OAT4 was significantly increased during PTL with and without inflammation. OAT-B was unaffected by either disease or labor. OATP-E mRNA was increased in samples from women with either PT-SPE or PTL-I. CYP19 (also known as aromatase) was increased in placenta from women with either PT-SPE or PTL-I. Estrogen receptor expressions (ER&.␣ and ER) were not different among groups, though there was an increasing trend observed with PTL-NI/PTL-I. Estrogen is known to up-regulate HSD11B2, which was also increased in samples from women with PTL with and without inflammation; CONCLUSION: The delivery of sulfated steroid precursors from the fetal blood to the placenta is important for estrogen synthesis and the normal progression of pregnancy. The increase of both steroid sulfate transporters (OAT-4 and OATP-E) and related enzymes (CYP19, HSD11B2) during various preterm-related disorders suggests a potential up-regulation of the estrogen biosynthetic machinery in early pregnancy that can be altered by pregnancy related disease states such as preeclampsia and inflammation. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.078

0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.076 56 54

PREDICTION OF SPONTANEOUS PRETERM BIRTH AND VERY PRETERM BIRTH IN TWIN PREGNANCIES USING SERIAL FETAL FIBRONECTIN AND CERVICAL LENGTH NATHAN FOX1, DANIEL SALTZMAN2, CHAD KLAUSER2, DANIELLE PERESS3, CHRISTINE GUTIERREZ4, ANDREI REBARBER2, 1Mount Sinai School of Medicine, Maternal Fetal Medicine, New York, New York, 2Mount Sinai School of Medicine, Maternal Fetal Medicine, New York, 3Mount Sinai School of Medicine, New York, 4New York Medical College, New York OBJECTIVE: To evaluate serial fetal fibronectin (FFN) and cervical length (CL) as predictors of spontaneous preterm birth (SPTB) in asymptomatic twin pregnancies STUDY DESIGN: Historical cohort of 168 twin pregnancies managed from 20052008 in a maternal-fetal medicine practice with serial (every 2-3 weeks) FFN and CL testing between 22 and 32 weeks gestation. Monoamniotic twins were excluded. A short CL was defined as ⬍20 mm. Iatrogenic preterm births were not included in the analysis. Chi square test, and chi square for trend were used when appropriate. A p-value of ⬍0.05 was considered significant. RESULTS: Any positive FFN from 22-32 weeks increased the risk of SPTB ⬍37 weeks (95% vs. 44%, p⬍.001), ⬍35 weeks (55% vs. 19.1%, p⫽.001), ⬍34 weeks (45% vs. 11.6%, p⫽.001), ⬍32 weeks (35% vs. 5%, p⬍.001), ⬍30 weeks (15% vs. 2.8%, p⫽.042), and ⬍28 weeks (15% vs. 2.1%, p⫽.026). Any CL⬍20mm from 22-32 weeks increased the risk of SPTB ⬍37 weeks (82.6% vs. 48.2%, p⫽.003), ⬍34 weeks (36% vs. 11.5%, p⫽.005), and ⬍32 weeks (26.9% vs. 4.5%, p⫽.001). The combination of a positive FFN and CL⬍20mm had a significantly higher positive predictive value than either positive test alone (Table 1). CONCLUSION: In twin pregnancies, either serial FFN testing or serial CL testing between 22 and 32 weeks can predict preterm birth and very preterm birth with a high positive predictive value. The combination of both tests significantly increases the positive predictive for preterm birth than either test alone. Consideration should be made for the routine use of these tests in all twin pregnancies to assess the risk of preterm birth. Risk of spontaneous preterm birth in twin pregnancies based on serial fetal fibronectin and cervical length tests from 22-32 weeks. Number % ⬍28w % ⬍30w % ⬍32w % ⬍34w % ⬍35w % ⬍37w All Negative 119 One Positive 24 Both Positive 11 p-value

1.7 8.3 18.2 .003

2.5 8.3 18.2 .012

4.3 8.3 54.5 ⬍.001

10.3 26.1 54.5 ⬍.001

18.4 39.1 54.5 .001

43.4 76.2 100 ⬍.001

DIVERSITY IN CYTOKINE SIGNATURE EXHIBITED BY BACTERIAL PATHOGENS ASSOCIATED WITH SPONTANEOUS PRETERM BIRTH IN HUMAN FETAL MEMBRANES STEPHEN FORTUNATO1, MORGAN PELTIER2, JUDITH ECKARDT3, RAMKUMAR MENON4, 1The Perinatal Research Center, Maternl-Fetal Medicine, Nashville, Tennessee, 2Department of Obstetrics, Gynecology and Reproductive Sciences University of Medicine and Dentistry of New Jersey, New Brunswick, New Jersey, 3The Perinatal Research Center, Nashville, Tennessee, 4Society for Maternal-Fetal Medicine, NAshville, Tennessee OBJECTIVE: Intraamniotic infection and cytokine mediated host inflammatory response is associated with spontaneous preterm birth (PTB). The inflammatory response produced by these bacteria may not be the same. This study documents differences in the cytokine signature of fetal membranes in response to four bacterial species involved in PTB. STUDY DESIGN: Fetal membranes (n⫽14) collected from cesareans at term (normal, not in labor) were placed in an organ explant system and were stimulated with heat inactivated suspensions of Ureaplasma parvum (UP), Gardanerella vaginalis (GV), E. coli, Group B Streptococcus (GBS) (107 colony forming units of each) for 24 hrs. Lipopolysaccharide (LPS ⫺ from E.coli O55:B5 ⫺100 ng/ml) stimulation was also performed for comparison to confirm membrane responsiveness. A panel of cytokines (IL-1␤, IL-6, IL-8, IL-10, TNF␣ and interferon [IFN]␥) was analyzed using multiplex assay. Kruskal Wallis test was used for statistical analysis. RESULTS: LPS increased the production of all cytokines from fetal membranes compared to unstimulated controls. E.coli stimulation, as expected, resembled that of LPS. Cytokine profiles were similar after GV and GBS stimulation where all cytokines except IL-8 and IFN␥ were higher than controls. UP stimulation, in keeping with the organisms of low virulence, demonstrated the least immune response with only IL-10 and TNF␣ concentrations being higher than controls. IFN␥ was stimulated only E.coli. IL-1␤, IL-6 and TNF␣ concentrations were several folds higher after E.coli stimulation compared to other three bacteria. CONCLUSION: The cytokine signature produced by four different bacterial species associated with PTB is distinctly different. E.coli shows the maximum proinflammatory effects and Ureaplasma parvum shows the least. The study shows that the pathways and the biomarkers involved in PTB may not be generalizable and even infection, the best understood of the etiologic factors, is not homogeneous in its presentation. This is suggestive of need for targeted interventions to effectively change the outcome. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.079

0002-9378/$ - see front matter doi:10.1016/j.ajog.2008.09.077

Supplement to DECEMBER 2008 American Journal of Obstetrics & Gynecology

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