1. Evaluation of Pregnancy PBMC and Placental MtDNA in HIV-infected, HAART-treated women, compared to HIV-uninfected women* Hélène Côté* 1,2, Evelyn Maan 3, Eszter Papp 1, Tessa Chaworth-Musters 3, Izabelle Gadawski 1, Julie van Schalkwyk 3, Marissa Jitratkosol 3, John Forbes 2,3, David Burdge 2,3, Deborah Money 2,3, and The HIV Perinatal Study Group 1 University of British Columbia; 2 Women`s Health Research Institute; and 3 Oak Tree Clinic / Children`s & Women`s Health Centre of BC, Vancouver, British Columbia, Canada 948 Hélène C.F. Côté helene.cote@ubc.ca Ph: 604-822-9777 FAX: 604-822-7635 HIV-infected women receive HAART during pregnancy, either from the 2 nd trimester until delivery, or throughout the pregnancy if indicated by CD4 count. NRTI can cause mitochondrial toxicity and are known to cross the placenta. This study evaluated longitudinal peripheral blood mononucleated cells (PBMC) and placental tissue mitochondrial DNA (mtDNA) levels in HIV-positive pregnancies compared to control HIV-negative pregnancies, as well as whole blood mtDNA levels in the infants at birth. Background Prospective single centre cohort study. Blood was collected in CPT tubes (BD) at 4 time periods in all pregnancies: 13-23w of gestation (before HAART for Group A), >23-30w, >30-39w, delivery (when possible) and for HIV pregnancies, at 6w post-partum. PBMC were isolated and washed in PBS 3 times at low speed. Whole blood was collected from infants (PKU heel prick) within 3 days of birth. Placental tissue was collected from fetal and maternal sides shortly after delivery. MtDNA content was measured in isolated maternal PBMC, infant whole blood and placental tissue by real-time PCR. Statistical analyses: Between group comparisons were done using the Mann-Whitney test. For placenta maternal vs. fetal side analyses, Wilcoxon signed-rank test and Pearson’s correlation were used. Methods Oak Tree Clinic The Perinatal HIV Study Group also includes: Ariane Alimenti Results Group A: N=27 for longitudinal PBMC, N=30 for placenta, HIV+ on HAART since 2 nd trimester of pregnancy (mostly AZT/3TC/nelfinavir HAART regimen) Group B: N=8 for longitudinal PBMC, 7 for placenta, HIV+ on HAART since conception (regimen varied) Group C: N=24 for PBMC, 23 for placenta: HIV negative controls Table 1. Placental mtDNA content. Figure 1. Longitudinal mean ± standard error maternal PBMC mtDNA content during pregnancy. Conclusions Acknowledgements A physiological increase in PBMC mtDNA is seen in normal pregnancy near the end of gestation, possibly to meet high energy demands. This increase appears delayed in HIV+ HAART-treated pregnancies. In HIV+ HAART-treated pregnancies, placental mtDNA tends to be higher on the maternal side compared to the fetal side of the organ. Changes in placental mtDNA may affect metabolism and energy production within the placenta. Infant blood mtDNA at birth content was not different between groups. Mitochondrial toxicity related to nucleoside therapy may have amplified consequences in a perinatal setting. Further studies are needed to identify antiretrovirals with the least toxicity in pregnancy. Figure 2. Comparison of placental mtDNA content between groups A and C on the maternal and fetal side. Funding to D. Money from: Figure 4. Correlation between placenta mtDNA content, maternal vs. fetal side. 0 50 100 150 200 250 300 050100150200250300 maternal side mtDNA/nDNA fetal side mtDNA/nDNA Group C = HIV- controls 0 50 100 150 200 250 300 050100150200250300 maternal side mtDNA/nDNA fetal side mtDNA/nDNA Group A = HIV+ HAART-exposed GroupNMATERNALFETALpaired comparisons Rslopep value A3097 ± 790 ± 70.4760.5040.0080.052 B781 ± 884 ± 150.7190.3590.0690.938 C2384 ± 885 ± 100.8991.05<0.00010.223 mean ± standard error Pearson's correlations Placenta mtDNA/nDNAmaternalvs. fetal A N=30 0 50 100 150 200 250 Placenta maternal side C N=23 mtDNA/nDNA P=0.058 Figure 3. Comparison of infant blood mtDNA content between groups at birth (0-3 days of age). 0 50 100 150 200 250 mtDNA/nDNA Placenta fetal side A N=30 C N=23 P=0.563 * Values presented here are slightly different from those in the abstract as a study participant originally been placed in Group B belonged in Group A 50 100 150 200 250 300 350 Blood mtDNA/nDNA A N=26 C N=19 B N=8 P=0.285 P=0.121P=0.518