COMBINED EFFECT OF ANTIBODIES TO BENZO[A]PYRENE, ESTRADIOL AND PROGESTERONE UPON SEX HORMONE CONCENTRATIONS IN BLOOD SERUM OF PREGNANT WOMEN WITH CONGENITAL MALFORMATIONS OF FETUS

Specific antibodies against environmental chemical gene toxicants and endogenous steroid hormones are shown to modulate concentrations of these compounds in blood serum and their biological effects in experimental models. However, probable hazards of such antibodies in human teratogenesis are still unknown. In particular, potential correlations between specific serum antibodies, sex hormone levels in pregnant women, and congenital malformations in newborns are not clear. The aim of this study was to identify possible associations between occurrence of antibodies to benzo[a]pyrene, estradiol and progesterone (Bp, Es and Pg, respectively), and congenital malformations, and effects of these antibodies upon Es and Pg concentrations in blood serum of pregnant women. We have included into the study 182 women with normal pregnancy and 101 females with congenital malformations of fetus. A non-competitive solid phase immunoassay was performed using Bp, Es and Pg conjugated to bovine serum albumin as antigens. Es and Pg serum concentrations were measured using immunoassay test-systems of “Immunotech” (Moscow). Results: strong positive correlations were revealed between the levels of studied antibodies in the both groups. High IgA-Bp/IgA-Es (> 3) and IgA-Bp/IgA-Pg ratios (> 3) were associated with congenital malformations (OR = 2.2, p = 0.013 and OR = 6.8, p < 0.0001). Positive correlations were revealed between Pg/Es and IgA-Bp/IgA-Es (rS = 0.62, p < 0.0001), and IgA-Bp/IgA-Pg ratios (rS = 0.77, p < 0.0001) in cases with inborn malformations. Similar correlations were found for the women who had normal pregnancy (rS = 0.4, p = 0.0001, and rS = 0.23, p = 0.026, respectively). The Pg/Es proportion correlated with IgG-Bp/IgG-Es (rS = 0.46, p = 0.002), and with IgG-Bp/IgG-Pg ratio (rS = 0.5, p = 0.0009) in cases of malformations, but not in women with normal pregnancy. Conclusion: we have revealed novel associations between congenital malformations of fetus and ratios of IgA-Bp/IgA-Es, as well as IgA-Bp/IgA-Pg, like as positive correlations between hormonal Pg/Es proportions, and ratios of specific antibodies in pregnant women

ASSOCIATIONS OF POLYMORPHISMS IN CYTOKINE GENES WITH THE INFECTIOUS GENESIS Of RECCURENT MISCARRIAGE

The associations of polymorphisms in cytokine genes with the infectious form recurrent miscarriage were investigated. Have established, that at the women a genotype AA of a gene TNFA (rs1800629) and combination of genotypes *1/*1 genes IL1RN and GG of a gene IL6 (rs1800795) are involved in pathogenesis recurrent miscarriage

Study of relationships between HLA-G gene polymorphism, intrauterine infection and recurrent miscarriage in women

The relationship between the HLA-G gene polymorphism (rs41551813, rs12722477, rs41557518), intrauterine infection and recurrent miscarriage (RM) in women were studied. The case group consisted of 180 patients with RM, defined as two or more consecutive miscarriages (min = 2; max = 8) at up to 20 weeks of gestation, and with clinically confirmed pregnancies and non-viable fetuses. At the time of examination. the women were enrolled from the Genetic Counseling Center at the Kemerovo Regional Clinical Hospital, Kemerovo, Russia, and were not pregnant. Each patient underwent a gynecological examination. We excluded women with a history of medical abortion, birth, and ectopic pregnancies. In addition, we excluded women with endocrine (e.g. diabetes) disorders. To exclude other known causes of spontaneous abortion, the following tests were performed: ultrasound examination of pelvic organs, and karyotyping in women and men. The women’s mean age in the RM group, was 29.6±4.8 (SD) years. The control group comprised 408 fertile women. These women didn’t have a history of spontaneous abortion, or a family history of congenital malformations. They have born, at least, 1-2 healthy children. Women’s mean age at birth of last child was 26.8±5.2 (SD) years. Influence of the intrauterine infection was analyzed on the basis of laboratory tests. Diagnostics of bacterial vaginosis and vulvo-vaginal candidiasis by microscopic examination was conducted. Viral agent infections (herpes simplex virus type 2, cytomegalovirus, human papilloma virus type 16/18), Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, Gardnerella vaginalis and Trichomonas vaginalis were detected by enzyme-linked immunoassay and polymerase chain reaction (PCR). The data were obtained from the medical cards of the surveyed women. All the women gave a written informed consent before participating in the study. Typing of polymorphisms of Thr31Ser (rs41551813, HLA-G*01:03) in exon 2, Leu110Ile (rs12722477, HLA-G*01:04) and 1597 delС (rs41557518, HLA-G*01:05N) in exon 3 HLA-G genes were performed by realtime PCR followed by melting analysis. The study showed that the intrauterine infection was not a risk factor for RM (p = 0.30) in the examined women. It was found that the 110 Ile allele (HLA-G *01:04) was a risk factor for RM both in women with intrauterine infection [ORa = 4.50 (2.41-8.38), p = 2.09e-06], and in women without infection [ORa = 2.46 (1.44-4.21), p = 0.0009]. The cooperative influence of genetic and infections factors with the risk of RM in women was revealed [ORa+f = 3.50 (2.01-6.09), p = 8.78e-06]. Our results will be useful in understanding the molecular mechanisms of immune disorders in fetomaternal interface, and for choosing the strategy of management and treatment in women with RM

ASSOCIATION BETWEEN CYTOKINE GENE POLYMORPHISMS AND RECURRENT MISCARRIAGE

Associations between IL1B (rs1143634), IL1RN (VNTR, intron 2), IL4 (VNTR, intron 3), TNFA (rs1800629, rs361525), IL6 (rs1800795), and IL10 (rs1800896) genetic polymorphisms in women with recurrent miscarriage (RM) were analyzed. We studied DNA samples of 112 women with RM and 267 women with physiological pregnancy. The IL1RN, IL4 genotypes were identified by PCR techniques, the IL1B, IL6 gene polymorphisms were defined by means of RFLP approach. To detect TNFA and IL10 gene polymorphisms, TaqMan real-time PCR was used. The results have shown that polymorphic loci of IL1B, IL1RN, IL10, TNFA genes were not associated with RM, and early spontaneous abortion risk. The 2R allele of IL4 gene was found to be associated with higher RM risk (OR = 1.52; 95% CI = [1.08-2.14]; P-value (cor) = 0.05), and G allele of IL6 gene was associated with a risk for > 3 early spontaneous abortions (OR = 2.10; 95% CI = [1.24-3.56]; P-value (cor) = 0.05), in an additive inheritance model. Upon evaluation of the data obtained, one may conclude that the IL4 (VNTR intron 3) and IL6 (rs1800795) gene polymorphisms could influence the RM development. These results may be useful for assessment of molecular mechanisms underlying early spontaneous abortion