evaluation of pregnancy pbmc and placental mtdna in hiv-infected,
DESCRIPTION
Oak Tree Clinic. Placenta maternal side. Placenta fetal side. P=0.058. P=0.563. 250. 250. 300. 300. A = HAART 2. A= HAART 2. trimester. trimester. nd. nd. 200. 200. B= HAART conception. B = HAART at conception. 250. 250. mtDNA/nDNA. C = HIV. C= HIV. -. -. controls. - PowerPoint PPT PresentationTRANSCRIPT
Evaluation of Pregnancy PBMC and Placental MtDNA in HIV-infected, HAART-treated women, compared to HIV-uninfected women*
Hélène Côté*1,2, Evelyn Maan3, Eszter Papp1, Tessa Chaworth-Musters3, Izabelle Gadawski1, Julie van Schalkwyk3, Marissa Jitratkosol3, John Forbes2,3, David Burdge2,3, Deborah Money2,3, and The HIV Perinatal Study Group
1University of British Columbia; 2Women`s Health Research Institute; and 3 Oak Tree Clinic / Children`s & Women`s Health Centre of BC, Vancouver, British Columbia, Canada
948Hélène C.F. Côté[email protected]: 604-822-9777FAX: 604-822-7635
HIV-infected women receive HAART during pregnancy, either from the 2nd 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
ResultsGroup A: N=27 for longitudinal PBMC, N=30 for placenta,
HIV+ on HAART since 2nd 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.
50
100
150
200
250
300
PB
MC
mtD
NA
/nD
NA
rati
o
13-23Weeks
>23-30Weeks
>30-39Weeks
Delivery 6 weeksPost-partum
C= HIV- controls
A= HAART 2nd trimester
B= HAART conception
Period in pregnancy
50
100
150
200
250
300
13-23Weeks
>23-30Weeks
>30-39Weeks
Delivery 6 weeksPost-partum
C = HIV - controls
A = HAART 2 nd trimester
B = HAART at conception
14
5
21
22
7
22
28
8
21
17
5
20
28
8
N
N
N
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 toD. Money from:
Figure 4. Correlation between placenta mtDNA content, maternal vs. fetal side.
0
50
100
150
200
250
300
0 50 100 150 200 250 300
maternal side mtDNA/nDNA
feta
l s
ide
mtD
NA
/nD
NA
Group C = HIV- controls
0
50
100
150
200
250
300
0 50 100 150 200 250 300
maternal side mtDNA/nDNA
feta
l s
ide
mtD
NA
/nD
NA
Group A = HIV+ HAART-exposed
Group N MATERNAL FETAL paired comparisons
R slope p value p value
A 30 97 ± 7 90 ± 7 0.476 0.504 0.008 0.052
B 7 81 ± 8 84 ± 15 0.719 0.359 0.069 0.938
C 23 84 ± 8 85 ± 10 0.899 1.05 <0.0001 0.223
mean ± standard error
Pearson's correlations
Placenta mtDNA/nDNA maternal vs . fetal
AN=30
0
50
100
150
200
250
Placenta maternal side
CN=23
mtD
NA
/nD
NA
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
mtD
NA
/nD
NA
Placenta fetal side
AN=30
CN=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
Blo
od
mtD
NA
/nD
NA
AN=26
CN=19
BN=8
P=0.285
P=0.121 P=0.518