- Email: [email protected]
931: Withdrawn
Poster Session V 931 Withdrawn 932 Withdrawn 933 Maternal peripheral blood mononuclear cells a novel source of sFlt-1 in preeclampsia pathology Stephen K. Gonzales, Martina Badell, Hanh Cottrell, Hongyan Qu, Neil Sidell, Augustine Rajakumar Emory University, Atlanta, GA
OBJECTIVE: Preeclampsia (PE) is a condition unique to pregnancy, and presents with a variable phenotypic expression in the mother and fetus. Increased plasma levels of anti-angiogenic VEGF receptor protein (sFlt-1) has been implicated in the pathogenesis of PE. The placenta remains the main source of sFlt-1 during pregnancy and PE. Growing evidence suggests that additional sources of sFlt-1 exist, such as maternal peripheral blood mononuclear cells (PBMCs). We aim to further characterize the PBMC contribution in PE. STUDY DESIGN: Plasma sFlt-1 levels were measured by ELISA in normal pregnancies (NP) and PE pregnancies using 2 collection tubes, EDTA and CTAD. The CTAD tubes prevent platelet activation and the PBMC release of sFlt-1. In addition, we performed heparin agarose enrichment of sFlt1 with Western blot analysis. Co-culture experiments were performed by incubating placental villous explants and PBMCs. We measured the sFlt-1 mRNA levels by real time PCR to determine the effect of co-culture. Data are presented as mean and significant at p<0.05. RESULTS: Plasma levels of sFlt-1 in women with PE (n¼12) was significantly higher than the levels in women with NP (n¼6) for both EDTA and CTAD tubes (p<.05) estimated by ELISA. sFlt-1 levels in CTAD plasma were 15% lower than in EDTA plasma (p<0.05) indicating that maternal PBMCs contribute to sFlt-1 production in both NP (14.6%, p<0.05) and PE women (16.7%, p<0.05). This was confirmed by Western blot. Real time PCR estimation of sFlt-1 mRNA shows a 3-fold increase in PE PBMCs co-cultured with PE placental villous explants (p<0.05) compared to PBMCs cultured alone. CONCLUSION: Our data suggests that maternal PBMC and placental interaction is important in the overexpression of sFlt-1. Additionally, our data support that PBMCs contribute significantly to PE pathology. PE is a unique disease in that the mother exhibits the significant phenotype, despite the placenta as the main source of sFlt-1. We speculate that intervention targeting PBMCs rather than the placenta to control PE may be more beneficial to promote better pregnancy outcomes.
934 Association of pre-pregnancy and longitudinal change in angiotensin-II with preterm preeclampsia Victoria L. Chase1, Carole A. McBride1, Gary Badger2, Erin A. Morris1, Ira M. Bernstein1 1
University of Vermont Medical Center, Burlington, VT, 2University of Vermont College of Medicine, Burlington, VT
OBJECTIVE: In uncomplicated pregnancy there is up-regulation of the
renin angiotensin system (RAS) with increased angiotensin-II (A-II) production, but decreased sensitivity to its vasopressor effects. Preeclampsia is associated with lower circulating RAS components but higher sensitivity to A-II. Our aim was to compare A-II concentrations prior to pregnancy, and changes during pregnancy, between women who developed preterm preeclampsia (PPR) compared with other pregnancy outcomes. STUDY DESIGN: Fifty-five subjects were evaluated longitudinally. Thirty-four women experienced normal pregnancy (NL), 7 developed gestational hypertension (GHTN), 7 developed term
ajog.org preeclampsia (TPR) and 7 developed preterm preeclampsia (PPR). Plasma A-II levels were obtained prior to conception and between 30 and 32 weeks gestation. Analysis of covariance was performed with p<.05 accepted for statistical significance. Data are presented as mean standard deviation (SD). RESULTS: Pre-pregnancy A-II levels were significantly higher in women who went on to develop PPR (NL¼ 35.6 11, GHTN¼ 37.2 11, TPR¼ 39.3 18.8, PPR¼ 50.8 20.3 pg/mL) with p¼.05; PPR versus other outcomes p¼.007. Third trimester A-II levels were similar between all groups (NL¼ 70.2 21.4, GHTN 70.8 29.3, TPR 81.4 18.9, PPR 63.1 23.4 pg/mL) with p¼.5. The change in A-II level during pregnancy was significantly lower in PPR pregnancies compared to all other outcomes (p¼.01). CONCLUSION: The RAS is believed to play a role in the development of hypertensive disorders of pregnancy. Women who develop PPR have higher pre-pregnancy A-II levels and a smaller increase during pregnancy compared with women who other pregnancy outcomes. This is consistent with prior research identifying reduced A-II levels intrapartum in preterm preeclamptics.
935 Evidence that perinatal hypertension and inflammation leads to chronic inflammation, hypertension and neurological changes in an animal model of HELLP syndrome Cynthia Bean, Shauna-Kay Spencer, Teylor Bowles, Patrick B. Kyle, Michelle Y. Owens, Kedra Wallace University of Mississippi Medical Center, Jackson, MS
OBJECTIVE: Hemolysis elevated liver enzyme low platelet (HELLP) syndrome is a life threatening syndrome with high maternal morbidity and mortality. Women with a history of HELLP syndrome are reported to be at an increased risk of developing psychological disorders in the immediate post-partum period and adverse cardiovascular events. Therefore we hypothesized that hypertension and inflammation during pregnancies associated with angiogenic imbalance contributes to this risk. STUDY DESIGN: On gestational day (GD) 12, mini-osmotic pumps infusing sFlt-1 and sEng were placed into rats to induce HELLP syndrome. A subset of HELLP rats received 2mg/kg of Orencia (Abatacept) on GD 13 and all rats had their mini-osmotic pumps removed 12-24hrs post-delivery to remove the source of sFlt-1 and sEng. Mean arterial pressure was measured between postpartum day (PPD) 32-40, followed by Evan’s blue infusion, tissue collection and euthanization. RESULTS: Rats with a history of HELLP syndrome (n¼12) were significantly hypertensive (p¼0.02) and still had evidence of liver damage (p¼0.04) compared to NP rats (n¼10; Table 1). HELLP rats treated with Orencia (n¼9) to decrease inflammation during pregnancy had a significant decrease in mean arterial pressure (p¼0.01) compared to untreated rats (Table 1). Rats with a history of HELLP syndrome had increased blood brain barrier permeability (BBB) in the brainstem/cerebellum compared to NP rats (p¼0.04), however this was reversed by treatment with Orencia (p¼0.006). Rats with HELLP syndrome also had a significant increase in hepatic levels of interleukin-6 (p¼0.04) and interleukin-beta (p¼0.04). Results from rats treated with Orencia are pending. Table 1. Table 1. Normal Pregnant HELLP Lactate dehydrogenase (IU/mL)
808.1+133.8
Aspartate aminotransferase (IU/mL) 146.2+12.86
HELLP+Orencia
1558+383.1
985.4+164.7
213+28.9*
161.5+11.8
Platelets (µL)
4.7x105+3.3x104
4.9x105+2.3x104 4.1x105+2.2x104
Mean arterial pressure (mmHg)
115.3+7.4
136.8+4.9*
S530 American Journal of Obstetrics & Gynecology Supplement to JANUARY 2017
117.2+4.7*
OBJECTIVE: Preeclampsia (PE) is a condition unique to pregnancy, and presents with a variable phenotypic expression in the mother and fetus. Increased plasma levels of anti-angiogenic VEGF receptor protein (sFlt-1) has been implicated in the pathogenesis of PE. The placenta remains the main source of sFlt-1 during pregnancy and PE. Growing evidence suggests that additional sources of sFlt-1 exist, such as maternal peripheral blood mononuclear cells (PBMCs). We aim to further characterize the PBMC contribution in PE. STUDY DESIGN: Plasma sFlt-1 levels were measured by ELISA in normal pregnancies (NP) and PE pregnancies using 2 collection tubes, EDTA and CTAD. The CTAD tubes prevent platelet activation and the PBMC release of sFlt-1. In addition, we performed heparin agarose enrichment of sFlt1 with Western blot analysis. Co-culture experiments were performed by incubating placental villous explants and PBMCs. We measured the sFlt-1 mRNA levels by real time PCR to determine the effect of co-culture. Data are presented as mean and significant at p<0.05. RESULTS: Plasma levels of sFlt-1 in women with PE (n¼12) was significantly higher than the levels in women with NP (n¼6) for both EDTA and CTAD tubes (p<.05) estimated by ELISA. sFlt-1 levels in CTAD plasma were 15% lower than in EDTA plasma (p<0.05) indicating that maternal PBMCs contribute to sFlt-1 production in both NP (14.6%, p<0.05) and PE women (16.7%, p<0.05). This was confirmed by Western blot. Real time PCR estimation of sFlt-1 mRNA shows a 3-fold increase in PE PBMCs co-cultured with PE placental villous explants (p<0.05) compared to PBMCs cultured alone. CONCLUSION: Our data suggests that maternal PBMC and placental interaction is important in the overexpression of sFlt-1. Additionally, our data support that PBMCs contribute significantly to PE pathology. PE is a unique disease in that the mother exhibits the significant phenotype, despite the placenta as the main source of sFlt-1. We speculate that intervention targeting PBMCs rather than the placenta to control PE may be more beneficial to promote better pregnancy outcomes.
934 Association of pre-pregnancy and longitudinal change in angiotensin-II with preterm preeclampsia Victoria L. Chase1, Carole A. McBride1, Gary Badger2, Erin A. Morris1, Ira M. Bernstein1 1
University of Vermont Medical Center, Burlington, VT, 2University of Vermont College of Medicine, Burlington, VT
OBJECTIVE: In uncomplicated pregnancy there is up-regulation of the
renin angiotensin system (RAS) with increased angiotensin-II (A-II) production, but decreased sensitivity to its vasopressor effects. Preeclampsia is associated with lower circulating RAS components but higher sensitivity to A-II. Our aim was to compare A-II concentrations prior to pregnancy, and changes during pregnancy, between women who developed preterm preeclampsia (PPR) compared with other pregnancy outcomes. STUDY DESIGN: Fifty-five subjects were evaluated longitudinally. Thirty-four women experienced normal pregnancy (NL), 7 developed gestational hypertension (GHTN), 7 developed term
ajog.org preeclampsia (TPR) and 7 developed preterm preeclampsia (PPR). Plasma A-II levels were obtained prior to conception and between 30 and 32 weeks gestation. Analysis of covariance was performed with p<.05 accepted for statistical significance. Data are presented as mean standard deviation (SD). RESULTS: Pre-pregnancy A-II levels were significantly higher in women who went on to develop PPR (NL¼ 35.6 11, GHTN¼ 37.2 11, TPR¼ 39.3 18.8, PPR¼ 50.8 20.3 pg/mL) with p¼.05; PPR versus other outcomes p¼.007. Third trimester A-II levels were similar between all groups (NL¼ 70.2 21.4, GHTN 70.8 29.3, TPR 81.4 18.9, PPR 63.1 23.4 pg/mL) with p¼.5. The change in A-II level during pregnancy was significantly lower in PPR pregnancies compared to all other outcomes (p¼.01). CONCLUSION: The RAS is believed to play a role in the development of hypertensive disorders of pregnancy. Women who develop PPR have higher pre-pregnancy A-II levels and a smaller increase during pregnancy compared with women who other pregnancy outcomes. This is consistent with prior research identifying reduced A-II levels intrapartum in preterm preeclamptics.
935 Evidence that perinatal hypertension and inflammation leads to chronic inflammation, hypertension and neurological changes in an animal model of HELLP syndrome Cynthia Bean, Shauna-Kay Spencer, Teylor Bowles, Patrick B. Kyle, Michelle Y. Owens, Kedra Wallace University of Mississippi Medical Center, Jackson, MS
OBJECTIVE: Hemolysis elevated liver enzyme low platelet (HELLP) syndrome is a life threatening syndrome with high maternal morbidity and mortality. Women with a history of HELLP syndrome are reported to be at an increased risk of developing psychological disorders in the immediate post-partum period and adverse cardiovascular events. Therefore we hypothesized that hypertension and inflammation during pregnancies associated with angiogenic imbalance contributes to this risk. STUDY DESIGN: On gestational day (GD) 12, mini-osmotic pumps infusing sFlt-1 and sEng were placed into rats to induce HELLP syndrome. A subset of HELLP rats received 2mg/kg of Orencia (Abatacept) on GD 13 and all rats had their mini-osmotic pumps removed 12-24hrs post-delivery to remove the source of sFlt-1 and sEng. Mean arterial pressure was measured between postpartum day (PPD) 32-40, followed by Evan’s blue infusion, tissue collection and euthanization. RESULTS: Rats with a history of HELLP syndrome (n¼12) were significantly hypertensive (p¼0.02) and still had evidence of liver damage (p¼0.04) compared to NP rats (n¼10; Table 1). HELLP rats treated with Orencia (n¼9) to decrease inflammation during pregnancy had a significant decrease in mean arterial pressure (p¼0.01) compared to untreated rats (Table 1). Rats with a history of HELLP syndrome had increased blood brain barrier permeability (BBB) in the brainstem/cerebellum compared to NP rats (p¼0.04), however this was reversed by treatment with Orencia (p¼0.006). Rats with HELLP syndrome also had a significant increase in hepatic levels of interleukin-6 (p¼0.04) and interleukin-beta (p¼0.04). Results from rats treated with Orencia are pending. Table 1. Table 1. Normal Pregnant HELLP Lactate dehydrogenase (IU/mL)
808.1+133.8
Aspartate aminotransferase (IU/mL) 146.2+12.86
HELLP+Orencia
1558+383.1
985.4+164.7
213+28.9*
161.5+11.8
Platelets (µL)
4.7x105+3.3x104
4.9x105+2.3x104 4.1x105+2.2x104
Mean arterial pressure (mmHg)
115.3+7.4
136.8+4.9*
S530 American Journal of Obstetrics & Gynecology Supplement to JANUARY 2017
117.2+4.7*