Materials/Methods: We compared the structure of the endometrium and
integrin expression in stimulated and non-stimulated endometrium. Endo-
metrial biopsies were obtained from ovum donors after ovarian stimulation,
7 days after hCG administration. These were compared with a control group
of fertile women attending for sterilisation. A biopsy taken 7 days following
the detection of urinary LH surge.Sections were immediately frozen in
liquid nitrogen and stored at Ϫ20°C until analysis. Immunohistochemical
analysis employed a panel of 5 monoclonal antibodies to analyse the
concentrations of the
1 integrins. The biopsies were
ﬁrstly dated by assessing the stage of the glandular development. Concen-
tration of integrins in the glandular and luminal epithelium were measured
using the HSCORE.
Results: There was a delay in the development of the glandular epithe-
lium after COH. There was also a reduction of all three integrins in the
glandular epithelium of biopsies from the donor patients when compared to
the control patients (all p Ͻ 0.05). The expression of the
3 integrin was
also reduced in the luminal epithelium (P Ͻ 0.003).
Conclusions: There is a delay in the development of the glandular
epithelium after superovulation. However the main ﬁnding is the reduction
in the expression of all 3 integrins in the endometrium. These integrins are
thought essential for implantation but may be suppressed in IVF cycles due
to the high oestradiol levels. Their reduction may lead to impaired implan-
tation during IVF treatment. Further work needs to be done to assess if the
integrin expression can be manipulated by altering the oestradiol levels
during IVF treatment.
A polymorphism of the tryptophan hydroxylase gene is not associated
with idiopathic recurrent miscarriage. G. Unfried, K. Lessel, F. Nagele,
C. B. Tempfer, J. Huber. Univ of Vienna, Vienna, Austria.
Objective: Immunologic factors are believed to play a major etiologic
role in the pathogenesis of idiopathic recurrent miscarriage (IRM). We
investigated the relation between IRM and a polymorphism in intron 7 of
the tryptophan hydroxylase (TPH) gene, an immunologic modulator on the
Design: Prospective case control study.
Materials/Methods: We investigated 125 women with a history of 3 or
more consecutive pregnancy losses before 20 weeks gestation and 137
healthy controls with at least 2 live births and no history of pregnancy loss.
Peripheral venous puncture, DNA extraction, and polymerase chain reaction
(PCR), followed by restriction fragment length polymorphism (RFLP) anal-
ysis were used to genotype women for the presence of the A218C poly-
morphism of the TPH gene.
Results: Allele frequencies among women with IRM and controls were
32.4% and 38.7%, respectively, for allele A (wild type) and 67.6% and
61.3%, respectively, for allele C (mutant). No association between the
presence of allele C and IRM was found (P ϭ .3, odds ratio [OR] 1.31;
Conﬁdence Interval [CI] 0.93 to 1.87). Genotype frequencies were also not
signiﬁcantly different between the study group (C/C: 44.8%, A/C: 45.6%,
A/A: 9.6%) and the control group (C/C: 37.2%, A/C: 48.2%, A/A: 14.6%;
P ϭ .2). Between women with primary and secondary IRM, no statistically
signiﬁcant differences with respect to allelic frequencies were observed
(63% vs. 62% for allele C and 31% vs. 38% for allele A; P ϭ .3).
Conclusions: This is the ﬁrst report on a TPH gene polymorphism among
women with IRM, demonstrating that the investigated A218C polymor-
phism in intron 7 is not associated with IRM in a Caucasian population.
Identiﬁcation of a novel progesterone response element in the HLA-G
gene promoter region. S. M. Yie, R. Xiao, C. L. Librach. Dept of
Obstetrics and Gynecology, Sunnybrook & Women’s Coll Health Science
Ctr and Max Bell Research Wing, The Toronto Gen Hosp, Toronto, ON,
Objective: Many functional characteristics of human leukocyte antigen G
(HLA-G) support the notion that it plays an important role in maternal
immune tolerance to the fetus. Previously. we reported that HLA-G protein
and mRNA levels are enhanced by progesterone in vitro through activation
of HLA-G gene transcription. Since we could not ﬁnd any previously
reported progesterone response element (PRE) sequences within the HLA-G
gene promoter region, the objective of this study was to identify any novel
progesterone response elements in this region.
Design: Controlled experimental study.
Materials/Methods: Progesterone receptors were prepared from the
MCF-7 breast cancer cell line treated with RU486 in culture. A 458 bp
HLA-G gene promoter fragment that has previously been shown to increase
CAT reporter gene expression by progesterone was isolated from JEG-3 cell
genomic DNA. A PRE derived from the mouse mammary tumor virus
(MMTV) and potential PRE site oligonucleotides derived from the fragment
were synthesized. The interaction between the receptor and DNA was
analyzed by an electrophoretic mobility shift assay (EMSA) and a DNA
fragment binding ELISA.
Results: Progesterone receptors bound speciﬁcally to both of the 458 bp
HLA-G promoter fragment and the control MMVT PRE. One of the
oligonucleotide constructs competed for progesterone receptor binding with
both the 458 bp fragment and the MMTV PRE. This oligonucleotide had a
similar binding afﬁnity to the progesterone receptor as the MMTV PRE.
However, it only had a 60% sequence homology with the MMTV PRE.
Conclusions: We have identiﬁed a novel progesterone response element
in the HLA-G gene sequence that may be responsible for HLA-G gene
transcriptional regulation by progesterone. Therefore, one mechanism for
the immunomodulation by progesterone during pregnancy may be through
regulation of HLA-G expression via this newly identiﬁed PRE.
Obstetrical family histories of women with unexplained recurrent preg-
nancy loss. C. A. Clark, K. A. Spitzer, J. N. Nadler, D. Wong, C. A.
Laskin. START Reproductive Biology Unit, Toronto, ON, Canada.
Objective: To determine whether there is an increased incidence of
pregnancy loss in the mothers of women with unexplained recurrent preg-
nancy loss (RPL).
Design: Survey analysis of the family history in women with recurrent
pregnancy loss compared to a control population.
Materials/Methods: We examined the family obstetrical histories of a
population of women with RPL (n ϭ 380) which was deﬁned as at least 2
losses in the absence of hormonal, anatomic or genetic abnormalities. Each
completed a questionnaire which asked for both her own and her mother’s
obstetrical history. A control group of 140 women who were attending a
family practice clinic were asked to complete a family history questionnaire
Results: Table 1 below shows the number of losses in each group. The
mothers of the women with RPL had no higher prevalence of pregnancy
losses than mothers of an unselected group of women.
Family history in women with recurrent pregnancy loss.
History of loss
n ϭ 380
n ϭ 380
n ϭ 140
At least 1 loss 380 (100) 135 (35.5) 45 (32.1)
1 Loss 0 80 (21.1) 28 (20.0)
2 Losses 96 (25.3) 30 (7.9) 9 (6.4)
Ն3 Losses 284 (74.7) 25 (6.6) 8 (5.7)
Conclusions: Based upon our review of the obstetrical histories of 520
women, there does not appear to be any familial pattern to recurrent
TUESDAY, OCTOBER 23, 2001
ART: CLINICAL ART
A survey of visitors to a science centre and website regarding the use of
donor gametes. J. Jenkins, C. Kailasam, C. Ford, L. Corrigan, K. Sykes.
Univ of Bristol, Bristol, UK; At-Bristol Science Ctr, Bristol., UK.
Abstracts Vol. 76, No. 3, Suppl. 1, September 2001