429: programming of offspring adaptive immune response following maternal inflammation

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STUDY DESIGN: Secondary analysis of maternal and surgical risk fac- tors from a RCT comparing pre and postnatal surgery for MMC. PTD was defined as delivery 34.0 weeks. NH was defined as a very thin area, or partial or complete dehiscence of the prior hysterotomy site. RESULTS: 89 patients underwent prenatal surgery; PTD occurred in 38(43%). The hysterotomy site was evaluated in 88; NH was reported in 31 women (35%) with complete or partial dehiscence in 10 of them. Surgery times, both total (skin incision to closure) and uterine, were longer in those with PTD than those who delivered later. Use of Du- ragen in fetal repair, oligohydramnios and spontaneous membrane rupture (SROM) were also associated with an increased risk for PTD. In a logistic regression, surgery time but not Duragen use remained significant, with an increase in odds of 20% for each 10 minute in- crease in surgery time (adjusted OR 1.02, 95% CI 1.00, 1.04). Total surgery time was also longer in those with subsequent oligohydram- nios (119.3 vs 100.9 minutes, p0.04) and SROM (112.5 vs 98.6 min- utes, p0.01) than those without, suggesting a causal pathway to PTD. NH occurred over twice as often in nulliparous as in multipa- rous patients and was associated with the need for longer duration of post operative MgSO4. Excluding the very thin cases, partial/complete dehiscence was also associated with these factors, and with oligohy- dramnios (p0.03). CONCLUSION: Longer surgical time is associated with the development of SROM and subsequent early preterm delivery. Development of oligohydramnios also places patients at increased risk for preterm delivery. Nulliparous patients should be counseled about the higher risk of hysterotomy complications. 428 Intracranial stem cell transplantation with and without erythropoietin facilitates motor recovery in a perinatal rat brain injury model Martin Mueller 1 , Andreina Schoeberlein 1 , Ursula Reinhart 1 , Ruth Sager 1 , Marianne Messerli 1 , Daniel Surbek 2 1 University of Bern, Obstetrics and Gynecology, Bern, Switzerland, 2 University Hospital of Bern, Obstetrics and Gynecology, Bern, Switzerland OBJECTIVE: Peripartal brain injury in the premature infant leads to a large spectrum of clinical problems including lifelong neurodevelop- mental deficits. We examined intracranial mesenchymal stem cells (MSC) transplantation as a promising therapy to facilitate motor re- covery after perinatal hypoxic-ischemic brain damage with inflamma- tory component in a perinatal rat model. STUDY DESIGN: The study was conducted in a sham controlled design. Subcutaneous administration of LPS from E. coli followed by ligation of the left carotid artery and hypoxia (8% O2, 30min) was performed on rats on postnatal d4. On postnatal d11 mesenchymal stem cells (MSC) were injected into the left lateral ventricle and administration of erythropoietin (1000 U/kg bw, ip) on postnatal d11, d12 and d13 was conducted in separate therapy group. In brain sections, donor human MSC were detected and evaluated by histology and immuno- histochemistry (postnatal d25 and d40). Spastic paresis (SP) was eval- uated by footprint and walking pattern (postnatal d40). RESULTS: Injected donor-derived cells were detected in the lateral ven- tricle by immunohistochemistry in both therapeutic groups. Migra- tion in the ventricle and parenchyma was also present. MSC’s with unaltered morphology were detected in the cortex cerebri. Applica- tion of MSC’s and MSC’s and EPO resulted in alleviation of SP. CONCLUSION: This study demonstrates that intracerebral transplanta- tion of human mesenchymal stem cells in a complex rat model of perinatal brain damage results in incorporation of MSCs in the le- sioned brain area an reduction of cerebral palsy. 429 Programming of offspring adaptive immune response following maternal inflammation Michael Ross 1 , Nizar Khatib 2 , Zeev Weiner 3 , Yuval Ginsberg 2 , Nibal Awad 2 , Shay Arison 2 , Israel Thaler 2 , Joseph Itskovitz-Eldor 2 , Ron Beloosesky 1 1 LABioMed at Harbor-UCLA Med. Ctr., Obstetrics and Gynecology, Torrance, CA, 2 Rambam Medical Center, Obstetrics and Gynecology, Haifa, Israel, 3 Rambam Medical Center, Technion Israel Institute of Technology, Obstetrics and Gynecology, Haifa, Israel OBJECTIVE: Prenatal infection with gram negative bacteria is common during pregnancy. We have shown previously that acute maternal LPS exposure at E18 programs significant decrease the offspring innate immune response. The adaptive immune response is composed of B and T lymphocytes. T helper (CD4, CD8-) and T Cytotoxic (CD8, CD4-) compose the majority of T cells, and are essential for adequate immune responses. While T helper cells play an important role in establishing and maximizing the capabilities of the immune system, T Cytotoxic destroy virally infected cells and tumor cells. We sought to determine whether the adaptive immune response is programmed by maternal inflammation. STUDY DESIGN: Pregnant Sprague Dawley rats (n4) at 18 days gesta- tion received intraperitoneal (i.p.) injections of LPS (500 ug/kg) or saline (control). Male and female pups were delivered spontaneously y (e21) and allowed to mature until postnatal day 24 (p24). CD4 and CD8 lymphocytes, granulocytes, and macrophages expressing Toll like receptor 4 (TLR4) were characterized in the blood, spleen, and thymus of offspring pups by flow cytometry. RESULTS: Pups of LPS treated dams had significantly lower blood T Cytotoxic lymphocytes than pups of saline treated dams, though the same number of T helper lymphocytes. Pups of LPS dams had signif- icantly higher counts of blood CD4 TLR4 and CD8 TLR4 lymphocytes, TLR4 granulocytes, and a trend towards higher TLR4 macrophages. The same pattern was demonstrated in the spleen and thymus. CONCLUSION: The decrease in the T Cytotoxic lymphocyte in the off- spring of LPS exposed dams may increase their ability to cope with viral infection and potential tumor development. The increase in CD4 TLR4 lymphocytes may aid in anti-inflammatory responses and may explain the decrease in the proinflammatory cytokines pre- viously demonstrated in these offspring following exposure to LPS. These results suggest that maternal inflammation during the antenatal period may induce long term sequelae in the offspring innate immune response which may predispose to adult disease. 430 Natural history for pregnancies complicated by obstructive uropathy and normal amniotic fluid volume Nahla Khalek 1 , Mark Johnson 1 1 The Children’s Hospital of Philadelphia, The Center for Fetal Diagnosis and Treatment, Philadelphia, PA OBJECTIVE: There is a paucity of literature documenting the outcome of fetuses prenatally diagnosed with obstructive uropathy and normal amniotic fluid volume. Current practice excludes these fetuses as can- didates for vesicoamniotic shunt placement. The aim of this interim analysis is to provide data up to delivery of affected neonates. Poster Session III Doppler Assessment, Fetus, Prematurity www.AJOG.org S198 American Journal of Obstetrics & Gynecology Supplement to JANUARY 2012

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STUDY DESIGN: Secondary analysis of maternal and surgical risk fac-tors from a RCT comparing pre and postnatal surgery for MMC. PTDwas defined as delivery � 34.0 weeks. NH was defined as a very thinarea, or partial or complete dehiscence of the prior hysterotomy site.RESULTS: 89 patients underwent prenatal surgery; PTD occurred in38(43%). The hysterotomy site was evaluated in 88; NH was reportedin 31 women (35%) with complete or partial dehiscence in 10 of them.Surgery times, both total (skin incision to closure) and uterine, werelonger in those with PTD than those who delivered later. Use of Du-ragen in fetal repair, oligohydramnios and spontaneous membranerupture (SROM) were also associated with an increased risk for PTD.In a logistic regression, surgery time but not Duragen use remainedsignificant, with an increase in odds of 20% for each 10 minute in-crease in surgery time (adjusted OR 1.02, 95% CI 1.00, 1.04). Totalsurgery time was also longer in those with subsequent oligohydram-nios (119.3 vs 100.9 minutes, p�0.04) and SROM (112.5 vs 98.6 min-utes, p�0.01) than those without, suggesting a causal pathway toPTD. NH occurred over twice as often in nulliparous as in multipa-rous patients and was associated with the need for longer duration ofpost operative MgSO4. Excluding the very thin cases, partial/completedehiscence was also associated with these factors, and with oligohy-dramnios (p�0.03).CONCLUSION: Longer surgical time is associated with the developmentof SROM and subsequent early preterm delivery. Development ofoligohydramnios also places patients at increased risk for pretermdelivery. Nulliparous patients should be counseled about the higherrisk of hysterotomy complications.

428 Intracranial stem cell transplantation withand without erythropoietin facilitates motorrecovery in a perinatal rat brain injury modelMartin Mueller1, Andreina Schoeberlein1, Ursula Reinhart1,Ruth Sager1, Marianne Messerli1, Daniel Surbek2

1University of Bern, Obstetrics and Gynecology, Bern, Switzerland,2University Hospital of Bern, Obstetrics and Gynecology, Bern, SwitzerlandOBJECTIVE: Peripartal brain injury in the premature infant leads to alarge spectrum of clinical problems including lifelong neurodevelop-mental deficits. We examined intracranial mesenchymal stem cells(MSC) transplantation as a promising therapy to facilitate motor re-covery after perinatal hypoxic-ischemic brain damage with inflamma-tory component in a perinatal rat model.STUDY DESIGN: The study was conducted in a sham controlled design.Subcutaneous administration of LPS from E. coli followed by ligationof the left carotid artery and hypoxia (8% O2, 30min) was performedon rats on postnatal d4. On postnatal d11 mesenchymal stem cells(MSC) were injected into the left lateral ventricle and administrationof erythropoietin (1000 U/kg bw, ip) on postnatal d11, d12 and d13was conducted in separate therapy group. In brain sections, donorhuman MSC were detected and evaluated by histology and immuno-

histochemistry (postnatal d25 and d40). Spastic paresis (SP) was eval-uated by footprint and walking pattern (postnatal d40).RESULTS: Injected donor-derived cells were detected in the lateral ven-tricle by immunohistochemistry in both therapeutic groups. Migra-tion in the ventricle and parenchyma was also present. MSC’s withunaltered morphology were detected in the cortex cerebri. Applica-tion of MSC’s and MSC’s and EPO resulted in alleviation of SP.CONCLUSION: This study demonstrates that intracerebral transplanta-tion of human mesenchymal stem cells in a complex rat model ofperinatal brain damage results in incorporation of MSCs in the le-sioned brain area an reduction of cerebral palsy.

429 Programming of offspring adaptive immuneresponse following maternal inflammationMichael Ross1, Nizar Khatib2, Zeev Weiner3, YuvalGinsberg2, Nibal Awad2, Shay Arison2, Israel Thaler2,Joseph Itskovitz-Eldor2, Ron Beloosesky1

1LABioMed at Harbor-UCLA Med. Ctr., Obstetrics and Gynecology,Torrance, CA, 2Rambam Medical Center, Obstetrics and Gynecology,Haifa, Israel, 3Rambam Medical Center, Technion Israel Instituteof Technology, Obstetrics and Gynecology, Haifa, IsraelOBJECTIVE: Prenatal infection with gram negative bacteria is commonduring pregnancy. We have shown previously that acute maternal LPSexposure at E18 programs significant decrease the offspring innateimmune response. The adaptive immune response is composed of Band T lymphocytes. T helper (CD4�, CD8-) and T Cytotoxic (CD8�,CD4-) compose the majority of T cells, and are essential for adequateimmune responses. While T helper cells play an important role inestablishing and maximizing the capabilities of the immune system, TCytotoxic destroy virally infected cells and tumor cells. We sought todetermine whether the adaptive immune response is programmed bymaternal inflammation.STUDY DESIGN: Pregnant Sprague Dawley rats (n�4) at 18 days gesta-tion received intraperitoneal (i.p.) injections of LPS (500 ug/kg) orsaline (control). Male and female pups were delivered spontaneouslyy (e21) and allowed to mature until postnatal day 24 (p24). CD�4 andCD�8 lymphocytes, granulocytes, and macrophages expressing Tolllike receptor 4 (TLR4) were characterized in the blood, spleen, andthymus of offspring pups by flow cytometry.RESULTS: Pups of LPS treated dams had significantly lower blood TCytotoxic lymphocytes than pups of saline treated dams, though thesame number of T helper lymphocytes. Pups of LPS dams had signif-icantly higher counts of blood CD4� TLR4� and CD8� TLR4�lymphocytes, TLR4� granulocytes, and a trend towards higherTLR4� macrophages. The same pattern was demonstrated in thespleen and thymus.CONCLUSION: The decrease in the T Cytotoxic lymphocyte in the off-spring of LPS exposed dams may increase their ability to cope withviral infection and potential tumor development. The increase inCD4� TLR4� lymphocytes may aid in anti-inflammatory responsesand may explain the decrease in the proinflammatory cytokines pre-viously demonstrated in these offspring following exposure to LPS.These results suggest that maternal inflammation during the antenatalperiod may induce long term sequelae in the offspring innate immuneresponse which may predispose to adult disease.

430 Natural history for pregnancies complicated byobstructive uropathy and normal amniotic fluid volumeNahla Khalek1, Mark Johnson1

1The Children’s Hospital of Philadelphia, The Centerfor Fetal Diagnosis and Treatment, Philadelphia, PAOBJECTIVE: There is a paucity of literature documenting the outcomeof fetuses prenatally diagnosed with obstructive uropathy and normalamniotic fluid volume. Current practice excludes these fetuses as can-didates for vesicoamniotic shunt placement. The aim of this interimanalysis is to provide data up to delivery of affected neonates.

Poster Session III Doppler Assessment, Fetus, Prematurity www.AJOG.org

S198 American Journal of Obstetrics & Gynecology Supplement to JANUARY 2012