Antibodies against beta 2-glycoprotein I (beta 2-GP I) and their relationship to fetoplacental antigens.

Binding of beta 2-GP I to anionic phospholipids is thought to be the major antigen required in the reaction of anticardiolipin antibodies to phospholipids. The aim of this study was to investigate the changes of anti-beta 2-GP I IgG during the first and second trimester of pregnancy and the relationship between the levels of anti-beta 2-GP I and fetoplacental antigens and the correlation between anti-beta 2-GP I IgG and antibodies against oxidized low-density lipoprotein IgG (oLAb) in serum of pregnant women. We determined anticardiolipin antibodies (ACA) IgG and maternal serum levels of alpha 1-fetoprotein (AFP), human chorionic gonadotrophin (HCG) and trophoblast-specific beta 1-glycoprotein (SP1) in 204 pregnant women in the first and second trimester. From this group we selected 52 serum samples positive for ACA IgG and 16 samples negative for ACA IgG. In the samples of selected patients, the levels of anti-beta 2-GP I IgG and oLAb IgG were determined. Anti-beta 2-GP I IgG levels significantly decreased in the second trimester (6.2+/-9.3 U/ml, mean +/- S.D.) in comparison with the first trimester (8.3+/-10.4 U/ml) (p=0.05). Multiple of median (MoM) AFP correlated negatively but not significantly in the first trimester with anti-beta 2-GP I (r = -0.261, p = 0.12). In the second trimester this correlation was significantly negative (r = -0.278, p = 0.04). The Spearman correlation coefficients for MoM HCG and anti-beta 2-GP I were 0.158 for the first trimester and 0.174 for the second trimester. MoM SP1 also did not correlate significantly with anti-beta 2-GP I in both trimesters. The correlation between anti-beta 2-GP I IgG and oLAb IgG was not significant (r = -0.06). In the first trimester 40 % serum samples were positive for anti-beta 2-GP I IgG and negative for oLAb IgG or vice versa, while 60 % samples in the second trimester were positive only for one determined autoantibody. We can conclude that the levels of anti-beta 2-GP I IgG decrease during the second trimester probably as the result of the effects of some immunosuppressive agents associated with pregnancy. The finding of negative correlation between AFP and anti-beta 2-GP I suggests that anti-beta 2-GP I has an influence on fetus development.

Antibodies against oxidized low density lipoproteins in pregnant women.

Oxidized low density lipoproteins (oxLDL) formed in vivo induce a humoral immune response. Oxidative modification of LDL renders it immunogenic and a heterogeneous population of specific anti-oxLDL antibodies is produced. These antibodies could represent a biological marker of oxidative stress and serve as markers of atherosclerosis. Autoantibodies against oxLDL (oLAb) have been detected in human subjects practically of every age. oLAb also appear in the blood of pregnant women. Some studies have shown that the levels of antibodies to oxLDL were elevated in women with established preeclampsia. The present study was aimed to estimate the oLAb IgG levels in the first and second trimester of pregnancy. Furthermore, we estimated the correlation between maternal serum (MS) levels of oLAb and alpha-1-fetoprotein (MS AFP), human chorionic gonadotrophin (MS HCG) and trophoblast-specific-beta-1-glycoprotein (MS SP1), because these proteins are determined as a part of prenatal biochemical screening for fetal congenital abnormalities. Our study deals with the oLAb changes in women with pregnancy-induced hypertension. We also investigated the correlation between oLAb IgG and anticardiolipin antibodies IgG (ACA) in the serum of pregnant women. We examined 40 pregnant women attending Institute for Mother and Child Care for their antenatal care as outpatients. Routine blood samplings between the 9-13th week of pregnancy and 16-18th week of pregnancy were performed as a part of biochemical prenatal screening for fetal congenital abnormalities (Group 1). Their mean age was 27 +/- 4.1 years. Furthermore, we examined 26 women in the second or third trimester with pregnancy-induced hypertension (Group 2). Group 2 was compared with 49 pregnant women in the second or third trimester who were normotensive (Group 3). We used commercial standardized ELISA kits for determination of oLAb IgG, ACA IgG, MS AFP and MS HCG, MS SP1 was analyzed by single radial immunodiffusion. We did not find any differences in the levels of oLAb IgG in the first and second trimester in the women of Group 1. The correlation between oLAb and ACA IgG was not statistically significant (Spearman coefficient r=0.22, p=0.1). The correlation between oLAb IgG with MS AFP, MS HCG and MS SP1 was not statistically significant. Weak negative correlation for AFP and HCG was suggested both in the first and in the second trimester. The levels of oLAb IgG in the group of women with pregnancy-induced hypertension were significantly lower than in the group of normotensive women (348 +/- 388 U/ml v.s. 579 +/- 400 mU/ml, p<0.01). We can conclude that the levels of oLAb do not differ in the first and second trimester of gravidity. However, we cannot exclude the possible influence of an inverse relationship between oLAb IgG titers and the synthesis of fetoplacental antigens. This finding is important especially in the context of the results of prenatal biochemical screening. Pregnancy-induced hypertension is associated with lower levels of oLAb. Weak cross-reactivity between oLAb and anticardiolipin antibodies may exist but there is a possibility that there are two different populations of antibodies reacting with various antigens.

[PAPP-A in the first trimester of pregnancy]. / PAPP-A v prvním trimestru tehotenství.

OBJECTIVE: Pregnancy-associated plasma protein A has been reported to be low in Down syndrome affected pregnancies during the first trimester of pregnancy. The aim of this study was to determine preliminary the medians of pregnancy-associated plasma protein A (PAPP-A) in the first trimester of pregnancy and to compare PAPP-A with other biochemical markers used for biochemical prenatal screening. DESIGN: Retrospective study. SETTING: First Institute of Medical Chemistry and Biochemistry and Institute for Clinical Biochemistry, First Medical Faculty, Charles University. Institute for Care of Mother and Child, Prague. PATIENTS: One hundred forty one pregnant women, who undergo biochemical prenatal screening for chromosomal disorders between 7th and 13th week were studied. In addition six women in the second trimester and five women with twin pregnancies, two cases of trisomy 21 and one case of trisomy 18 in second trimester were available for study. METHODS: Maternal serum levels of PAPP-A, human chorionic gonadotropin (hCG) and alfa-1-feto-protein (AFP) were measured using ELISA methods. A single radial immunodiffusion was used to determine trophoblast-specific-beta-1-glycoprotein (SP1). RESULTS: PAPP-A levels increased throughout the first trimester with median 1.8 mg/l in the 7th week to 23.0 mg/l in the 13th week of pregnancy. PAPP-A serum levels from 3 women with twin pregnancies were higher than in women with singleton pregnancies. Serum levels of PAPP A in two women with fetus affected by chromosomal disorders did not differ from normal pregnancies. Correlation coefficients between PAPP-A and AFP and between PAPP-A and SP1 were statistically significant (r = 0.42, P < 0.001, respectively r = 0.54, P < 0.001). The levels of PAPP-A and HCG did not correlate significantly (r = 0.019, P = 0.82). CONCLUSION: We established first trimester medians for PAPP-A, which are necessary for evaluation of the pathological values. We found statistically significant correlation between PAPP-A and SP1 and PAPP-A and AFP.

Prevalence of various antiphospholipid antibodies in pregnant women.

Antiphospholipid antibodies (APAs) are characterized as a heterogeneous population of autoantibodies directed against different target antigens, predominantly anionic phospholipids or phospholipid-containing structures. The presence of APAs has been strongly associated with a variety of clinical disorders including adverse pregnancy complications such as spontaneous abortions, pregnancy-induced hypertension, preeclampsia and intrauterine growth retardation. The purpose of this study was to compare the prevalence of anticardiolipin antibodies (ACAs), which are routinely examined, with APAs directed against phosphatidylserine (APS), phosphatidylinositol (API), phosphatidylethanolamine (APE) and phosphatidylcholine (APC) in the sera of pregnant women. We examined 410 serum samples of pregnant women hospitalized in the department for pathological pregnancies. They underwent prenatal biochemical screening of fetal congenital abnormalities in the first and the second trimester of gravidity. Anticardiolipin IgG and IgM were measured using commercial ELISA kits (ImmuLisa Anti-Cardiolipin Antibody), whereas APS, APE, API and APC were determined by our modified ELISA kit. Among 410 pregnant women we found 21 patients (5.1%) positive for ACA IgG (>20 GPL) and 30 patients (7.3%) positive for ACA IgM (>10 MPL). It was found that 7.8% of pregnant women had at least one high-titer APA IgG and 9.8% high-titer APA IgM. One third of ACA IgG or IgM positive sera contained polyspecific autoantibodies reactive to at least two various phospholipids. In the group of IgG ACA positive women, 28.6% patients were positive for APS, 28.6% were positive or moderately positive for API, 23.8% for APC and 19% for APE. In the group of IgM ACA positive women, 33.3% were also positive for APS, 26.7% for APE, 26.7% for API and 23.3% for APC were present. IgG and IgM ACA negative patients exhibited a significantly lower incidence of other APA than the group of ACA positive pregnant women. It still remains to clarify if the routine examination of APA reacting with other anionic and zwitterionic antigens other than cardiolipin would improve the probability of identifying women liable to adverse pregnancy complications.

[Embryo-fetoscopy--present possibilities of endoscopy in obstetrics]. / Embryofetoskopie--soucasné moznosti endoskopie v porodnictví.

OBJECTIVE: The introduction of the embryofetoscopy--a new method of prenatal diagnosis used in the 1st trimester of the pregnancy. SETTING: Institute for the Care of Mother and Child, Prague. DESIGN: Pilot clinical study. METHODS: The embryofetoscopy was carried out in pregnant women in the 7th or 8th conceptional week who asked for a legal abortion of the pregnancy. We used the embryofetoscope with diameter of 0.8 mm (made by K. Storz, Tuttlingen, Germany). The trocar of the embryofetoscope was inserted into the animal cavity transabdominally always guided by ultrasound. It is possible to perform the examination in local anaesthesia. A further trocar with two canals--one for insertion of the embryofetoscope, the second of the needle with diameter of 0.6 mm--is designated for genetic indicated sampling of embryonal tissues or for incidental application of gene or stem cells therapy in the future. RESULTS: The videocamera documented our embryoscopical findings till this time in 7 cases. On the pictures 1-8 are the face, scull, eye, ear, extremities, genitals and umbilical cord of the embryo in the 7th conceptional week. CONCLUSIONS: The applications of the embryofetoscopy are very wide. It allows us to obtain not only new knowledge about the early development of the normal and affected embryo in vivo, but it simultaneously opens new ways for the therapy of the embryo in the future and for fetal surgery guided by embryoscope in the 2nd and 3rd trimester of the pregnancy.

[Comparative study of the occurrence of beta-2-glycoprotein I antibodies (anti-beta-2-GPI) and anticardiolipin antibodies (ACA) in the blood of pregnant women]. / Srovnávací studie výskytu protilátek proti beta-2-glykoproteinu I (anti-beta-2-GPI) a antikardiolipinových protilátek (ACA) v séru tehotných zen.

OBJECTIVE: It is accepted now that most antiphospholipid antibodies (APA) are not directed against phospholipids alone but complexes of phospholipid-binding proteins and phospholipids. One of the phospholipid-binding proteins is beta 2-glycoprotein I (beta 2-GPI), a normal plasma glycoprotein, which under physiological conditions binds with negative charged phospholipids. Measurement of anti-beta 2-GPI antibodies requires specific tests, since ELISA for determination of anticardiolipin antibodies (ACA) may detect both antibodies directly binding cardiolipin and antibodies against cardiolipin-binding proteins. In this study a comparison of APAs against cardiolipin (CL), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylethanolamine (PE) v.s. anti-beta 2-GPI were compared. SETTING: First Institute of Medical Chemistry and Biochemistry, First Medical Faculty, Charles University and Institute for Care of Mother and Child. DESIGN: Retrospective clinical study using stored sera for determination of APA. METHODS: One hundred twenty four women in the first and the second trimester, who undergo biochemical prenatal screening for chromosomal disorders by maternal serum alpha-fetoprotein (MS AFP), human chorionic gonadotrophin (MS HCG) and trophoblast-specific beta 1-glycoprotein (MS SP1) were studied. In serum samples antibodies against CL, PS, PE, PI (isotype IgG and IgM) were examined by solid ELISA. 19 women who were positive at least for one type of APAs were selected and in the serum samples the levels of anti-beta 2-GPI were measured. RESULTS: No pregnant woman with anti-beta 2-GPI had positive antibodies against other phospholipids except CL. There was no significant correlation between levels of ACA IgG and IgM v.s. anti-beta 2-GPI IgG and IgM. Eight percent of serum samples were positive for both anti-beta 2-GPI IgG and ACA IgG. In the second trimester a statistically significant decrease of ACA IgG was found (p = 0.014). The difference of ACA IgM and anti-beta 2-GPI IgG and IgM was not statistically significant. Levels of foetoplacental antigens in anti-beta 2-GPI positive pregnant women were normal. CONCLUSION: A correlation between anti-beta 2-GPI antibodies and ACA was not found in our study. Anti-beta 2-GPI antibodies and ACA may be two different subpopulations of APA.

[Anti-phosphatidylserine antibodies in women with reproductive disorders]. / Antifosfatidylserinové protilátky u zen s poruchami reprodukce.

Antiphosphatidylserine antibodies (APSA) belong to the heterogeneous population of antiphospholipid antibodies (APA) which are oriented above all against negatively charged phospholipids. The presence of APA in women is closely associated with repeated miscarriages and other complications during pregnancy. The most frequently detected specific antibodies in these patients are autoantibodies against cardiolipin and phosphatidylserine (PS). In a group of 84 pregnant women where within the framework of biochemical prenatal screening of inborn developmental defects serum levels of alpha-1-fetoprotein, choriogonadotropin and trophoblast specific beta-1-glycoprotein were examined as well as in 22 women treated for primary sterility and 22 blood donors the authors assessed, using the ELISA method, antiphosphatidylserine and cardiolipin antibodies (ACA). They found an increased prevalence of APSA in all examined groups as compared with the control group of blood donors. In pregnant women the prevalence of APSA and ACA did not differ and at least one type of antibodies was detected in 20.1%. In pregnant women with positive APSA in the case-records spontaneous abortions were recorded, or imminent abortions during the present gestation or treatment on account of sterility, and in some instances also changes of foetoplacental antigen serum levels were found. It is therefore likely that the presence of APA in women may be one of the factors participating in reproductive disorders and that assessment of APSA together with APA may extend the spectrum of immunological examinations, in particular in sterile and infertile women.

11 beta-hydroxyandrostenedione in human amniotic fluid.

11 beta-Hydroxyandrostene-3,17-dione (11 beta-hydroxyandrostenedione), a potential marker for prenatal diagnosis of CAH, has been for the first time determined in amniotic fluid obtained from 68 women who later delivered children without endocrine disorders. Its concentration averaged 0.96 +/- 0.31 (S.D.) (range 0.48-1.85) nmol/l and did not depend on the fetal sex. There was mild, insignificant increase of these values along with duration of pregnancy.