- Email: [email protected]
The impact of antiretroviral drugs in placental vascular endothelium dysfunction
Abstracts / Placenta 57 (2017) 225e335
vascular endothelial growth factor (VEGF) and ATP. We used qPCR with specific primers for Piezo1, using b-actin as a control. Results: Piezo1 mRNA was found consistently in snap frozen placental tissue from normal and FGR samples. Piezo1 was also expressed in cultured PlECs. In response to shear stress, PlECs aligned in the direction of flow. Applying shear stress across individual cell membranes induced a calcium entry response. Using short interfering RNA (siRNA) to deplete Piezo1 significantly reduced the response to shear stress. Conclusion: Human placental endothelium is responsive to shear stress and shows consistent Piezo1 expression. We hypothesise that in pregnancies complicated by FGR, alterations in shear stress will modulate MIC activation. Further studies into Piezo1 will provide increased understanding of blood flow regulation in the placenta, potentially resulting in new therapeutic targets for FGR.
273
Fig. 2. Graphs showing the average radius (left), distance (middle), and tortuosity (right) for central (blue) and eccentric (orange) cord insertion with increasing branch order. Vessel radius and distance are equivalent, but eccentric cord insertion is associated with less tortuous low order branches.
http://dx.doi.org/10.1016/j.placenta.2017.07.161
http://dx.doi.org/10.1016/j.placenta.2017.07.160
P1.74. QUANTIFYING THE STRUCTURE OF THE CHORIONIC VASCULAR TREE WITH CENTRAL AND ECCENTRIC CORD INSERTION Rosalind Pratt 1, 2, Andrew Melbourne 1, Ciaran Hutchinson 1, 3, Owen Arthurs 3,1, Neil J. Sebire 3,1, Tom Vercauteren 1, Sebastien Ourselin 1, Anna L. David 1, 2. 1 University College London, London, UK; 2 University College London Hospital, London, UK; 3 Great Ormond Street Hospital, London, UK Objectives: Marginal cord insertion is associated with poor pregnancy outcome, however the effect cord insertion site has on the chorionic vascular tree is unknown. We perform quantified surface vessel analysis to investigate the effect of cord insertion site. Methods: Six placentae were collected with consent at elective, term caesarean section. High resolution surface vessel photographs were taken following perfusion via an umbilical artery. The arterial tree was semiautomatically segmented. Cord insertion site was calculated as distance to closest placental edge/shortest placental diameter x 100. Central cord insertion was defined as greater than or equal to 33%, eccentric as less than 33%. Novel analysis algorithms written in MATLAB skeletonised the vascular tree, and analysis was performed as a function of the branching structure (Fig. 1). Results: Four placentae had central cord insertions (40.0%, 47.9%, 41.7%, 42.7%) and 2 had eccentric (25.3%, 22.4%). Vessel radius decreased and distance from cord insertion increased with branch order in both (Fig. 2A/ B). Tortuosity decreases with branch number with central cord insertion, but eccentric cord insertion was associated with less tortuous vessels proximally (Fig. 2C). Conclusions: Eccentric cord insertion is associated with measurably less tortuous vessels close to the cord insertion. Quantifying the vascular tree allows us to understand the extent of heterogeneity possible within healthy placenta, and the underlying aetology of pathology.
P1.75. EFFECTS OF INTERACTION
EV
MIRNAS
ON
TROPHOBLASTIC-ENDOTHELIAL
rrez-Samudio, Udo Markert, Diana Morales-Prieto. Placenta Ruby Gutie Labor, Jena, Thuringia, Germany Background: MicroRNAs are short non-coding RNAs involved in posttranscriptional regulation of gene expression. Pregnancy-associated miRNAs control trophoblast functions and can be delivered into maternal cells via Extracellular Vesicles (EVs) secretion. The present study explored the function of trophoblast EV-encapsulated miRNAs on the trophoblastendothelial cell interaction. Methods: Trophoblastic cell lines were transfected with miR-141 and miR519d mimics, or non-genomic controls. Transfected cells were cultured in medium supplemented with exosome-free fetal bovine serum for 24h before being seed for functional assays and their supernatants were recovered. EVs were isolated by ultracentrifugation, and characterized by Nanoparticle Tracking Analysis (NTA) and CD63 assessed by dot blotting. To mimic the trophoblast-endothelial cell interaction, JEG-3 and HTR8/ SVneo cells were co-cultured with human umbilical vein endothelial cells (HUVEC) in a 3D model. The ability of trophoblast cells to interact with HUVECs was examined by inversed microscopy and ImageJ analysis. Results: Overexpression of miR-141 in trophoblastic cells diminishes their ability to interact with HUVECs. Treatment with EV miR-141 on non-transfected cells has the same effect. Overexpression of miR-519d and treatment with EVs miR-519 increases HTR8/SVneo but not JEG-3 cell function. Conclusion: Placenta-associated miRNAs regulate trophoblast-endothelial cell interaction in vitro suggesting an important role on remodeling of spiral arteries in normal and pathological human pregnancies. http://dx.doi.org/10.1016/j.placenta.2017.07.162
P1.76. THE IMPACT OF ANTIRETROVIRAL DRUGS IN PLACENTAL VASCULAR ENDOTHELIUM DYSFUNCTION Onankoy Onyangunga, Jagidesa Moodley, Frederick Odun-Ayo, Thajasvarie Naicker. University of KwaZulu-Natal, Durban, KwaZulu-Natal, South Africa
Fig. 1. Example of a segmented, skeletonised chorionic vascular tree, showing the branch order (A). Segmented vascular tree of placenta with central (A-D) and eccentric (E-G) cord insertion showing segmented branches (B, E), branch radius (C,F), and branch tortuosity (D,G).
Objective: The introduction of antiretroviral drugs (ARV) in the treatment of HIV infection has reduced drastically the morbidity and mortality of HIV infected patients. However the success of the Highly Active Antiretroviral Therapy (HAART) has raised challenges with its side effects particularly on the cardiovascular system. There is a paucity of data on the vascular placental and foetal unit. The burden of HIV infection in South Africa with 31.5% of infected mothers needs to be addressed as regards the issues of foetal consequences of maternal ARV drugs. This study investigates the impact of ARVs on vascular endothelial cell in the placenta of normotensive HIV infected mothers with lymphatic vessel endothelial hyaluron receptor1(LYVE-1) marker.
274
Abstracts / Placenta 57 (2017) 225e335
Methods: Placental tissues obtained from normotensive HIV infected mothers (n¼30) on HAART regimen for at least for 3 months were immunostained using LYVE-1 primary monoclonal antibody. The patient were divided into four groups; each given different regimen. The expression of LYVE-1 in the different regimen and normotensive HIV uninfected (control) groups were analysed. Results: In regimen 1, five were on dual therapy Zidovudine. In regimen 2, seven were on Nevarapine + lamivudine + stavudine. In regimen 3 (nevirapine + lamivudine + tenofovir) and 4 (full drug combination; FDC), five and thirteen patients were noted. An up-regulation of LYVE-1 expression was observed in the HIV infected mothers with no statistical differences in the different regimens. Conclusion: Although the limitation of this study is small sample size. Hence, these preliminary results may require for large cohorts to assess further impact of other markers particularly those affecting the mitochondria and inflammatory factors in the placenta, foetus and neonate of HIV mothers. http://dx.doi.org/10.1016/j.placenta.2017.07.163
P1.77. FETAL SEX AFFECTS UTERINE NATURAL KILLER CELL ACTIVATION AND SPIRAL ARTERY ANGIOGENESIS IN A TRANSGENIC MOUSE MODEL OF INTRAUTERINE GROWTH RESTRICTION Jessica Hebert, Terry Morgan. OHSU, Portland, OR, USA Objectives: We have recently shown using 3D microcomputed tomography and quantitative microbubble enhanced ultrasound that mildly elevated maternal angiotensinogen (AGT) expression (1.2-fold in transgenic (TG) dams versus wild-type (WT) controls) is sufficient to significantly inhibit spiral artery growth during pregnancy, which leads to abnormal uteroplacental blood flow, placental damage, and fetal growth restriction. Male fetal sex is associated with less spiral artery branching in TG dams compared with their female siblings. Since uterine natural killer cells (uNK) play an important role in spiral artery angiogenesis, we hypothesized less uNK cell activation in TG dams, which may be accentuated in males. Methods: Whole mount uteroplacental units from WT and TG dams (3 litters per genotype) were harvested and stained for Ly49 and DBA at days 6.5 and 8.5 following a protocol published by the Croy laboratory (2012). Briefly, the number of Ly49 and DBA positive CD45 positive uNK cells were counted per high power field (20x objective) counting at least two metrial triangle fields per uteroplacental unit. Fetal sex was determined by SRYPCR using fetal tissue. Results: WT whole mount counts were similar to those reported by the Felker et al. (2016) in the Croy laboratory (Table). The Ly49/DBA ratio was higher in females than males at day 6.5. Male progeny from TG dams surprisingly had relatively fewer Ly49+ uNK cells at day 6.5 and fewer DBA+ uNK cells at day 8.5 compared with WT controls. Conclusion: Differences in maternal uterine spiral artery remodeling relative to fetal sex in our TG model of fetal growth restriction may be related to relatively fewer DBA+ activated uNK cells at day 8.5. Future studies will correlate these findings in a larger cohort with metrial triangle angiogenesis at day 8.5 and placental trophoblast invasion.
Mouse Gestational Age
Croy, 2016 M&F
WT Male
WT Female
TG Male
TG Female
Day 6.5 (Ly49/DBA)
1.9 ± 0.2 (reference)
3.2 ± 0.6 (n¼4)
1.9 ± 0.4 (n¼4)
2.0 ± 0.4 (n¼6)
1.0 ± 0.2* (n¼3)
Day 8.5 (Ly49/DBA)
0.8 ± 0.01 (reference)
0.8 ± 0.08 (n¼6)
0.9 ± 0.2 (n¼3)
ND
2.1 ± 0.2* (n¼3)
http://dx.doi.org/10.1016/j.placenta.2017.07.164
P1.78. ALTERED MATERNAL IMMUNE SYSTEM PROFILE IN POSTPARTUM PE: POTENTIAL CONTRIBUTION TO PROGRESSION
PE AND DISEASE
Marie-eve Brien 1, 2, Cyntia Duval 1, 2, Ines Boufaied 1, 2, Mubina Juvanovic 1, 2, Dorothee Dal Soglio 1, 2, Sylvie Girard 1, 2. 1 STE-Justine Hospital Research Center, Montreal, Quebec, Canada; 2 Universite de Montreal, Montreal, Quebec, Canada Preeclampsia (PE) is a leading cause of mortality/morbidity which affects 5-8% of pregnancies. PE is characterized by de novo hypertension and the placenta has a central role in the pathophysiology with its removal being the only cure. However, it cannot explain PE occurring in the postpartum period (PPPE). Systemic inflammation is known to be associated with PE and could be key to understand PPPE. Objective: We aimed to determine the circulating inflammatory profile in women with PE or PPPE as compared to uncomplicated term pregnancies in relation with maternal-fetal interface inflammation. Methods: We recruited women with uncomplicated term pregnancies (Ctrl, N¼20), pregnancies with PE (PE, N¼20) or normal pregnancies with postpartum PE (PPPE, N¼20). Blood samples were collected to determine the circulating immune profile and placental biopsies obtained for histological and inflammatory mediators analysis. Results: Maternal circulating immune cells profiles were distinct between PE and PPPE and characterized by significant increase in the percentage of CD8+ and CD4+ (p<0.01) cells in PE and increased NK cells and monocytes (p<0.01) in PPPE. Circulating levels of endogenous mediators of inflammation also differed with elevated uric acid in PE only (p<0.01) and increased HMGB1 solely in PPPE (p<0.05). At the maternal-fetal interface increased syncytial knots were observed in PE without morphological changes were observed in PPPE. Interestingly, high levels of immune cell infiltration were detected in placentas from women that later developed PPPE (p<0.001). Conclusion: Our work showed unique maternal immune profiles in PE vs PPPE which highlight the involvement of the maternal immune system in both pathologies. Furthermore, we saw immune cells infiltration in the placenta of women that will later develop PPPE suggesting a prenatal initiation of the pathology. In-depth analysis of these changes will allow better understanding of the mechanisms linking the immune system to PE and PPPE. http://dx.doi.org/10.1016/j.placenta.2017.07.165
P1.79. REDUCED MITOCHONDRIAL RESPIRATION IN PLACENTAS FROM EARLYONSET PRE-ECLAMPSIA: POTENTIAL ROLES OF THE MITOCHONDRIAL UNFOLDED PROTEIN RESPONSE Emilie Dommett 1, Francesca Colleoni 1, John Kingdom 2, Matts Andrew Murray 1, Graham Burton 1, Hong Wa Olovsson 3, Yung 1. 1 University of Cambridge, Cambridge, UK; 2 University of Toronto, Toronto, Canada; 3 Uppsala University, Uppsala, Sweden Objectives: Mitochondrial dysfunction has been observed in placentas from cases of pre-eclampsia. Our group has demonstrated the pathophysiological role of placental endoplasmic reticulum unfolded protein response (UPRER) in the aetiology of early- (EOPE), but not late-onset (LOPE) pre-eclampsia (Yung et al 2014). As there is close linkage between the UPRER and mitochondrial unfolded protein response (UPRmt), we investigated potential roles of UPRmt in regulation of mitochondrial function. Methods: Placental samples were obtained with ethical approval from 7 normotensive controls, 7 EOPE (<34 wk) and 7 LOPE (>34 wk) cases. Activity of mitochondrial electron transport chain complexes (ETC) was measured by respirometry using Clarke electrodes. Levels of ETC complexes (Oxphos antibodies); UPRmt molecular markers including TID1, HSP60, GRP75, Clpp and, Paraplegin; and citrate synthase (mitochondrial
vascular endothelial growth factor (VEGF) and ATP. We used qPCR with specific primers for Piezo1, using b-actin as a control. Results: Piezo1 mRNA was found consistently in snap frozen placental tissue from normal and FGR samples. Piezo1 was also expressed in cultured PlECs. In response to shear stress, PlECs aligned in the direction of flow. Applying shear stress across individual cell membranes induced a calcium entry response. Using short interfering RNA (siRNA) to deplete Piezo1 significantly reduced the response to shear stress. Conclusion: Human placental endothelium is responsive to shear stress and shows consistent Piezo1 expression. We hypothesise that in pregnancies complicated by FGR, alterations in shear stress will modulate MIC activation. Further studies into Piezo1 will provide increased understanding of blood flow regulation in the placenta, potentially resulting in new therapeutic targets for FGR.
273
Fig. 2. Graphs showing the average radius (left), distance (middle), and tortuosity (right) for central (blue) and eccentric (orange) cord insertion with increasing branch order. Vessel radius and distance are equivalent, but eccentric cord insertion is associated with less tortuous low order branches.
http://dx.doi.org/10.1016/j.placenta.2017.07.161
http://dx.doi.org/10.1016/j.placenta.2017.07.160
P1.74. QUANTIFYING THE STRUCTURE OF THE CHORIONIC VASCULAR TREE WITH CENTRAL AND ECCENTRIC CORD INSERTION Rosalind Pratt 1, 2, Andrew Melbourne 1, Ciaran Hutchinson 1, 3, Owen Arthurs 3,1, Neil J. Sebire 3,1, Tom Vercauteren 1, Sebastien Ourselin 1, Anna L. David 1, 2. 1 University College London, London, UK; 2 University College London Hospital, London, UK; 3 Great Ormond Street Hospital, London, UK Objectives: Marginal cord insertion is associated with poor pregnancy outcome, however the effect cord insertion site has on the chorionic vascular tree is unknown. We perform quantified surface vessel analysis to investigate the effect of cord insertion site. Methods: Six placentae were collected with consent at elective, term caesarean section. High resolution surface vessel photographs were taken following perfusion via an umbilical artery. The arterial tree was semiautomatically segmented. Cord insertion site was calculated as distance to closest placental edge/shortest placental diameter x 100. Central cord insertion was defined as greater than or equal to 33%, eccentric as less than 33%. Novel analysis algorithms written in MATLAB skeletonised the vascular tree, and analysis was performed as a function of the branching structure (Fig. 1). Results: Four placentae had central cord insertions (40.0%, 47.9%, 41.7%, 42.7%) and 2 had eccentric (25.3%, 22.4%). Vessel radius decreased and distance from cord insertion increased with branch order in both (Fig. 2A/ B). Tortuosity decreases with branch number with central cord insertion, but eccentric cord insertion was associated with less tortuous vessels proximally (Fig. 2C). Conclusions: Eccentric cord insertion is associated with measurably less tortuous vessels close to the cord insertion. Quantifying the vascular tree allows us to understand the extent of heterogeneity possible within healthy placenta, and the underlying aetology of pathology.
P1.75. EFFECTS OF INTERACTION
EV
MIRNAS
ON
TROPHOBLASTIC-ENDOTHELIAL
rrez-Samudio, Udo Markert, Diana Morales-Prieto. Placenta Ruby Gutie Labor, Jena, Thuringia, Germany Background: MicroRNAs are short non-coding RNAs involved in posttranscriptional regulation of gene expression. Pregnancy-associated miRNAs control trophoblast functions and can be delivered into maternal cells via Extracellular Vesicles (EVs) secretion. The present study explored the function of trophoblast EV-encapsulated miRNAs on the trophoblastendothelial cell interaction. Methods: Trophoblastic cell lines were transfected with miR-141 and miR519d mimics, or non-genomic controls. Transfected cells were cultured in medium supplemented with exosome-free fetal bovine serum for 24h before being seed for functional assays and their supernatants were recovered. EVs were isolated by ultracentrifugation, and characterized by Nanoparticle Tracking Analysis (NTA) and CD63 assessed by dot blotting. To mimic the trophoblast-endothelial cell interaction, JEG-3 and HTR8/ SVneo cells were co-cultured with human umbilical vein endothelial cells (HUVEC) in a 3D model. The ability of trophoblast cells to interact with HUVECs was examined by inversed microscopy and ImageJ analysis. Results: Overexpression of miR-141 in trophoblastic cells diminishes their ability to interact with HUVECs. Treatment with EV miR-141 on non-transfected cells has the same effect. Overexpression of miR-519d and treatment with EVs miR-519 increases HTR8/SVneo but not JEG-3 cell function. Conclusion: Placenta-associated miRNAs regulate trophoblast-endothelial cell interaction in vitro suggesting an important role on remodeling of spiral arteries in normal and pathological human pregnancies. http://dx.doi.org/10.1016/j.placenta.2017.07.162
P1.76. THE IMPACT OF ANTIRETROVIRAL DRUGS IN PLACENTAL VASCULAR ENDOTHELIUM DYSFUNCTION Onankoy Onyangunga, Jagidesa Moodley, Frederick Odun-Ayo, Thajasvarie Naicker. University of KwaZulu-Natal, Durban, KwaZulu-Natal, South Africa
Fig. 1. Example of a segmented, skeletonised chorionic vascular tree, showing the branch order (A). Segmented vascular tree of placenta with central (A-D) and eccentric (E-G) cord insertion showing segmented branches (B, E), branch radius (C,F), and branch tortuosity (D,G).
Objective: The introduction of antiretroviral drugs (ARV) in the treatment of HIV infection has reduced drastically the morbidity and mortality of HIV infected patients. However the success of the Highly Active Antiretroviral Therapy (HAART) has raised challenges with its side effects particularly on the cardiovascular system. There is a paucity of data on the vascular placental and foetal unit. The burden of HIV infection in South Africa with 31.5% of infected mothers needs to be addressed as regards the issues of foetal consequences of maternal ARV drugs. This study investigates the impact of ARVs on vascular endothelial cell in the placenta of normotensive HIV infected mothers with lymphatic vessel endothelial hyaluron receptor1(LYVE-1) marker.
274
Abstracts / Placenta 57 (2017) 225e335
Methods: Placental tissues obtained from normotensive HIV infected mothers (n¼30) on HAART regimen for at least for 3 months were immunostained using LYVE-1 primary monoclonal antibody. The patient were divided into four groups; each given different regimen. The expression of LYVE-1 in the different regimen and normotensive HIV uninfected (control) groups were analysed. Results: In regimen 1, five were on dual therapy Zidovudine. In regimen 2, seven were on Nevarapine + lamivudine + stavudine. In regimen 3 (nevirapine + lamivudine + tenofovir) and 4 (full drug combination; FDC), five and thirteen patients were noted. An up-regulation of LYVE-1 expression was observed in the HIV infected mothers with no statistical differences in the different regimens. Conclusion: Although the limitation of this study is small sample size. Hence, these preliminary results may require for large cohorts to assess further impact of other markers particularly those affecting the mitochondria and inflammatory factors in the placenta, foetus and neonate of HIV mothers. http://dx.doi.org/10.1016/j.placenta.2017.07.163
P1.77. FETAL SEX AFFECTS UTERINE NATURAL KILLER CELL ACTIVATION AND SPIRAL ARTERY ANGIOGENESIS IN A TRANSGENIC MOUSE MODEL OF INTRAUTERINE GROWTH RESTRICTION Jessica Hebert, Terry Morgan. OHSU, Portland, OR, USA Objectives: We have recently shown using 3D microcomputed tomography and quantitative microbubble enhanced ultrasound that mildly elevated maternal angiotensinogen (AGT) expression (1.2-fold in transgenic (TG) dams versus wild-type (WT) controls) is sufficient to significantly inhibit spiral artery growth during pregnancy, which leads to abnormal uteroplacental blood flow, placental damage, and fetal growth restriction. Male fetal sex is associated with less spiral artery branching in TG dams compared with their female siblings. Since uterine natural killer cells (uNK) play an important role in spiral artery angiogenesis, we hypothesized less uNK cell activation in TG dams, which may be accentuated in males. Methods: Whole mount uteroplacental units from WT and TG dams (3 litters per genotype) were harvested and stained for Ly49 and DBA at days 6.5 and 8.5 following a protocol published by the Croy laboratory (2012). Briefly, the number of Ly49 and DBA positive CD45 positive uNK cells were counted per high power field (20x objective) counting at least two metrial triangle fields per uteroplacental unit. Fetal sex was determined by SRYPCR using fetal tissue. Results: WT whole mount counts were similar to those reported by the Felker et al. (2016) in the Croy laboratory (Table). The Ly49/DBA ratio was higher in females than males at day 6.5. Male progeny from TG dams surprisingly had relatively fewer Ly49+ uNK cells at day 6.5 and fewer DBA+ uNK cells at day 8.5 compared with WT controls. Conclusion: Differences in maternal uterine spiral artery remodeling relative to fetal sex in our TG model of fetal growth restriction may be related to relatively fewer DBA+ activated uNK cells at day 8.5. Future studies will correlate these findings in a larger cohort with metrial triangle angiogenesis at day 8.5 and placental trophoblast invasion.
Mouse Gestational Age
Croy, 2016 M&F
WT Male
WT Female
TG Male
TG Female
Day 6.5 (Ly49/DBA)
1.9 ± 0.2 (reference)
3.2 ± 0.6 (n¼4)
1.9 ± 0.4 (n¼4)
2.0 ± 0.4 (n¼6)
1.0 ± 0.2* (n¼3)
Day 8.5 (Ly49/DBA)
0.8 ± 0.01 (reference)
0.8 ± 0.08 (n¼6)
0.9 ± 0.2 (n¼3)
ND
2.1 ± 0.2* (n¼3)
http://dx.doi.org/10.1016/j.placenta.2017.07.164
P1.78. ALTERED MATERNAL IMMUNE SYSTEM PROFILE IN POSTPARTUM PE: POTENTIAL CONTRIBUTION TO PROGRESSION
PE AND DISEASE
Marie-eve Brien 1, 2, Cyntia Duval 1, 2, Ines Boufaied 1, 2, Mubina Juvanovic 1, 2, Dorothee Dal Soglio 1, 2, Sylvie Girard 1, 2. 1 STE-Justine Hospital Research Center, Montreal, Quebec, Canada; 2 Universite de Montreal, Montreal, Quebec, Canada Preeclampsia (PE) is a leading cause of mortality/morbidity which affects 5-8% of pregnancies. PE is characterized by de novo hypertension and the placenta has a central role in the pathophysiology with its removal being the only cure. However, it cannot explain PE occurring in the postpartum period (PPPE). Systemic inflammation is known to be associated with PE and could be key to understand PPPE. Objective: We aimed to determine the circulating inflammatory profile in women with PE or PPPE as compared to uncomplicated term pregnancies in relation with maternal-fetal interface inflammation. Methods: We recruited women with uncomplicated term pregnancies (Ctrl, N¼20), pregnancies with PE (PE, N¼20) or normal pregnancies with postpartum PE (PPPE, N¼20). Blood samples were collected to determine the circulating immune profile and placental biopsies obtained for histological and inflammatory mediators analysis. Results: Maternal circulating immune cells profiles were distinct between PE and PPPE and characterized by significant increase in the percentage of CD8+ and CD4+ (p<0.01) cells in PE and increased NK cells and monocytes (p<0.01) in PPPE. Circulating levels of endogenous mediators of inflammation also differed with elevated uric acid in PE only (p<0.01) and increased HMGB1 solely in PPPE (p<0.05). At the maternal-fetal interface increased syncytial knots were observed in PE without morphological changes were observed in PPPE. Interestingly, high levels of immune cell infiltration were detected in placentas from women that later developed PPPE (p<0.001). Conclusion: Our work showed unique maternal immune profiles in PE vs PPPE which highlight the involvement of the maternal immune system in both pathologies. Furthermore, we saw immune cells infiltration in the placenta of women that will later develop PPPE suggesting a prenatal initiation of the pathology. In-depth analysis of these changes will allow better understanding of the mechanisms linking the immune system to PE and PPPE. http://dx.doi.org/10.1016/j.placenta.2017.07.165
P1.79. REDUCED MITOCHONDRIAL RESPIRATION IN PLACENTAS FROM EARLYONSET PRE-ECLAMPSIA: POTENTIAL ROLES OF THE MITOCHONDRIAL UNFOLDED PROTEIN RESPONSE Emilie Dommett 1, Francesca Colleoni 1, John Kingdom 2, Matts Andrew Murray 1, Graham Burton 1, Hong Wa Olovsson 3, Yung 1. 1 University of Cambridge, Cambridge, UK; 2 University of Toronto, Toronto, Canada; 3 Uppsala University, Uppsala, Sweden Objectives: Mitochondrial dysfunction has been observed in placentas from cases of pre-eclampsia. Our group has demonstrated the pathophysiological role of placental endoplasmic reticulum unfolded protein response (UPRER) in the aetiology of early- (EOPE), but not late-onset (LOPE) pre-eclampsia (Yung et al 2014). As there is close linkage between the UPRER and mitochondrial unfolded protein response (UPRmt), we investigated potential roles of UPRmt in regulation of mitochondrial function. Methods: Placental samples were obtained with ethical approval from 7 normotensive controls, 7 EOPE (<34 wk) and 7 LOPE (>34 wk) cases. Activity of mitochondrial electron transport chain complexes (ETC) was measured by respirometry using Clarke electrodes. Levels of ETC complexes (Oxphos antibodies); UPRmt molecular markers including TID1, HSP60, GRP75, Clpp and, Paraplegin; and citrate synthase (mitochondrial