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13: Inhibition of toll-like receptors and inflammatory response in primary chorion cells
IDSOG Abstracts
ajog.org
Figure Infectivity of CVL fluid from pregnant and non-pregnant women expressed as a percentage of inhibition compared to positive and negative (no cell) controls
13 Inhibition of toll-like receptors and inflammatory response in primary chorion cells J. Thompson1, L. Feng2, B. Antczak2, P. Seed3, A. Murtha2 1
Vanderbilt University, Department of Ob/Gyn, 2Duke University, Department of Ob/Gyn, 3Duke University, Department of Pediatrics
OBJECTIVES: Activation of toll-like receptors (TLR) result in production of inflammatory cytokines and proteases that play a critical role in the pathogenesis of preterm birth (PTB). Infection with Ureaplasma spp. is a risk factor for adverse pregnancy outcomes including preterm premature rupture of membranes and PTB. The objective of this study was to determine if chorion epithelial cell exposure to Ureaplasma parvum induced inflammation and if this response is mediated through TLR2 and/or TLR6. METHODS: Primary chorion cells were harvested from four subjects undergoing elective term uncomplicated cesarean deliveries prior to the onset of labor or membrane rupture. Chorion cells were stimulated with Ureaplasma parvum in the presence and absence of inhibitors to TLR2 and TLR6. Cells were collected and RNA extracted using the RNeasyÒ minikit. RNA concentration was measured and reverse transcribed into complementary DNA and real time PCR performed using probes against IL-8. Duplicate samples were evaluated with average IL-8 cycle thresholds (CT) normalized to beta 2 microglobulin. IL-8 expression is expressed as fold change compared to the untreated control or the Ureaplasma treated group. Data were evaluated by ANOVA and Mann-Whitney. RESULTS: Ureaplasma significantly increased IL-8 expression compared to untreated controls (2.9 fold increase) and TLR inhibitors (TLR 2 and/or 6) did not alter IL-8 expression (TLR2 inhibitor 0.8 fold change; TLR6 inhibitor 0.8 fold change; TLR2+TLR6 inhibitors 0.9 fold change). Inhibition of TLR2 and TLR6 resulted in a significant decrease in Ureaplasma parvum induced IL-8 expression (inhibition TLR2 p¼0.03; inhibition TLR6 p¼0.03). Combination of TLR2 + TLR6 inhibition did not significantly decrease IL-8 expression although there was a trend toward significance (p¼0.057). CONCLUSIONS: Ureaplasma induced inflammation appears to be mediated through TLR2 and TLR6. Inhibition of TLR2 and TLR6 significantly decreases Ureaplasma parvum induced IL-8 expression in primary chorion cells. LEARNING OBJECTIVE: identify the relationship between Ureaplasma induced inflammation and its relationship with TLR2 and TLR6.
14 Uptake of a third trimester HIV testing program at a tertiary care hospital Erica J. Hardy1,2, Sarah Atunah-Jay2, Brian Chow3, Robin Neale1, Susan Cu-Uvin4, Brenna Hughes1,2 1 Women & Infants Hospital, Providence, RI, 2Alpert Medical School of Brown University, 3Department of Pediatrics, Marshfield Clinic, Wisconsin, 4 Miriam Hospital, Providence, RI
OBJECTIVES: To estimate the difference in third trimester HIV testing at a tertiary care hospital after implementation of a third trimester testing program, and to identify factors associated with third trimester HIV testing after the implementation. METHODS: A retrospective study of prenatal records from two cohorts of women: those delivering in 2011 or 2013, before and after the implementation of third trimester testing program. Delivery records were randomly selected by prenatal care location in equal numbers from each care location. Inclusion criteria included full term delivery, initiation of prenatal care before 24 weeks gestation and no transfer of care after 24 weeks gestation. HIV testing required documentation on a primary lab report or medical record. Women with HIV testing performed at the time of labor and delivery were excluded. Covariates associated with HIV infection and thought to impact testing practices were measured. RESULTS: After the implementation of the testing program, HIV testing in the third trimester increased significantly from 3% tested to 49% (p<0.0001). After controlling for factors that were different between the two cohorts, HIV testing remained significantly higher after program implementation (aOR: 81, 95% CI: 25-253). Other factors that remained associated with receiving a third trimester HIV test were fewer prenatal visits (OR 0.88, 95%CI 0.81-0.98), and having a third trimester STD test (OR 7.43, 95%CI 3.04-18.15). Among the 2013 cohort, factors that associated with third trimester HIV testing were prenatal care location (p<0.0001), with 3% of those who received care in a private office receiving a third trimester test compared to 71% from a public health clinic, and 84% from the hospital-based clinic. Also associated with increased rates of HIV testing were younger maternal age (median age of 26 vs. 30, p¼0.019), fewer prenatal visits (mean of 10 vs. 12, p¼0.024), nonwhite race (p<0.0001), Hispanic ethnicity (p<0.0001), initiation of prenatal care in the second trimester (p¼0.001) and having a third trimester STD test (p<0.0001). CONCLUSIONS: Implementation of a third trimester testing program was effective in increasing third trimester HIV testing rates, however even after program implementation testing rates were significantly associated with location of prenatal care, maternal age, parity, race, ethnicity, insurance type, and receipt of a third trimester STD test. LEARNING OBJECTIVE: Learners will be able to identify factors that are associated with third trimester HIV testing rates after the implementation of a third trimester HIV testing program.
15 Relationship of specific bacteria in the cervical and vaginal microbiotas with cervicitis L. Sycuro1, L. Gorgos2, S. Srinivasan1, T. Fiedler1, M. Morgan1, J. Balkus1, R. S. McClelland3,4, D. Fredricks1,3, J. Marrazzo3 1
Fred Hutchinson Cancer Research Center, Seattle, WA, 2Special Immunology Associates, El Rio Health Center, Tucson, AZ, 3University of Washington, Seattle, WA, 4University of Nairobi, Nairobi, Kenya
OBJECTIVES: Cervicitis is an inflammatory condition of the cervix
associated with upper genital tract infection and reproductive DECEMBER 2015 American Journal of Obstetrics & Gynecology
901
ajog.org
Figure Infectivity of CVL fluid from pregnant and non-pregnant women expressed as a percentage of inhibition compared to positive and negative (no cell) controls
13 Inhibition of toll-like receptors and inflammatory response in primary chorion cells J. Thompson1, L. Feng2, B. Antczak2, P. Seed3, A. Murtha2 1
Vanderbilt University, Department of Ob/Gyn, 2Duke University, Department of Ob/Gyn, 3Duke University, Department of Pediatrics
OBJECTIVES: Activation of toll-like receptors (TLR) result in production of inflammatory cytokines and proteases that play a critical role in the pathogenesis of preterm birth (PTB). Infection with Ureaplasma spp. is a risk factor for adverse pregnancy outcomes including preterm premature rupture of membranes and PTB. The objective of this study was to determine if chorion epithelial cell exposure to Ureaplasma parvum induced inflammation and if this response is mediated through TLR2 and/or TLR6. METHODS: Primary chorion cells were harvested from four subjects undergoing elective term uncomplicated cesarean deliveries prior to the onset of labor or membrane rupture. Chorion cells were stimulated with Ureaplasma parvum in the presence and absence of inhibitors to TLR2 and TLR6. Cells were collected and RNA extracted using the RNeasyÒ minikit. RNA concentration was measured and reverse transcribed into complementary DNA and real time PCR performed using probes against IL-8. Duplicate samples were evaluated with average IL-8 cycle thresholds (CT) normalized to beta 2 microglobulin. IL-8 expression is expressed as fold change compared to the untreated control or the Ureaplasma treated group. Data were evaluated by ANOVA and Mann-Whitney. RESULTS: Ureaplasma significantly increased IL-8 expression compared to untreated controls (2.9 fold increase) and TLR inhibitors (TLR 2 and/or 6) did not alter IL-8 expression (TLR2 inhibitor 0.8 fold change; TLR6 inhibitor 0.8 fold change; TLR2+TLR6 inhibitors 0.9 fold change). Inhibition of TLR2 and TLR6 resulted in a significant decrease in Ureaplasma parvum induced IL-8 expression (inhibition TLR2 p¼0.03; inhibition TLR6 p¼0.03). Combination of TLR2 + TLR6 inhibition did not significantly decrease IL-8 expression although there was a trend toward significance (p¼0.057). CONCLUSIONS: Ureaplasma induced inflammation appears to be mediated through TLR2 and TLR6. Inhibition of TLR2 and TLR6 significantly decreases Ureaplasma parvum induced IL-8 expression in primary chorion cells. LEARNING OBJECTIVE: identify the relationship between Ureaplasma induced inflammation and its relationship with TLR2 and TLR6.
14 Uptake of a third trimester HIV testing program at a tertiary care hospital Erica J. Hardy1,2, Sarah Atunah-Jay2, Brian Chow3, Robin Neale1, Susan Cu-Uvin4, Brenna Hughes1,2 1 Women & Infants Hospital, Providence, RI, 2Alpert Medical School of Brown University, 3Department of Pediatrics, Marshfield Clinic, Wisconsin, 4 Miriam Hospital, Providence, RI
OBJECTIVES: To estimate the difference in third trimester HIV testing at a tertiary care hospital after implementation of a third trimester testing program, and to identify factors associated with third trimester HIV testing after the implementation. METHODS: A retrospective study of prenatal records from two cohorts of women: those delivering in 2011 or 2013, before and after the implementation of third trimester testing program. Delivery records were randomly selected by prenatal care location in equal numbers from each care location. Inclusion criteria included full term delivery, initiation of prenatal care before 24 weeks gestation and no transfer of care after 24 weeks gestation. HIV testing required documentation on a primary lab report or medical record. Women with HIV testing performed at the time of labor and delivery were excluded. Covariates associated with HIV infection and thought to impact testing practices were measured. RESULTS: After the implementation of the testing program, HIV testing in the third trimester increased significantly from 3% tested to 49% (p<0.0001). After controlling for factors that were different between the two cohorts, HIV testing remained significantly higher after program implementation (aOR: 81, 95% CI: 25-253). Other factors that remained associated with receiving a third trimester HIV test were fewer prenatal visits (OR 0.88, 95%CI 0.81-0.98), and having a third trimester STD test (OR 7.43, 95%CI 3.04-18.15). Among the 2013 cohort, factors that associated with third trimester HIV testing were prenatal care location (p<0.0001), with 3% of those who received care in a private office receiving a third trimester test compared to 71% from a public health clinic, and 84% from the hospital-based clinic. Also associated with increased rates of HIV testing were younger maternal age (median age of 26 vs. 30, p¼0.019), fewer prenatal visits (mean of 10 vs. 12, p¼0.024), nonwhite race (p<0.0001), Hispanic ethnicity (p<0.0001), initiation of prenatal care in the second trimester (p¼0.001) and having a third trimester STD test (p<0.0001). CONCLUSIONS: Implementation of a third trimester testing program was effective in increasing third trimester HIV testing rates, however even after program implementation testing rates were significantly associated with location of prenatal care, maternal age, parity, race, ethnicity, insurance type, and receipt of a third trimester STD test. LEARNING OBJECTIVE: Learners will be able to identify factors that are associated with third trimester HIV testing rates after the implementation of a third trimester HIV testing program.
15 Relationship of specific bacteria in the cervical and vaginal microbiotas with cervicitis L. Sycuro1, L. Gorgos2, S. Srinivasan1, T. Fiedler1, M. Morgan1, J. Balkus1, R. S. McClelland3,4, D. Fredricks1,3, J. Marrazzo3 1
Fred Hutchinson Cancer Research Center, Seattle, WA, 2Special Immunology Associates, El Rio Health Center, Tucson, AZ, 3University of Washington, Seattle, WA, 4University of Nairobi, Nairobi, Kenya
OBJECTIVES: Cervicitis is an inflammatory condition of the cervix
associated with upper genital tract infection and reproductive DECEMBER 2015 American Journal of Obstetrics & Gynecology
901