Immunochemical study of the specific antigens of human amniotic fluid]

Four specific antigens (trophoblastic beta 1-globulin, placental lactogen, alpha 1- and alpha 2-globulins of human placenta) were identified using antisera to the native amniotic fluid. Five antigens with the mobility of prealbumins, alpha 1-globulins, alpha 2-globulins and beta 2-globulins which bear no resemblance with the previously studied antigens were identified using antisera to the acid fraction of the amniotic fluid. Both the prealbumins and alpha 2-globulin were found in the blood serum of foetuses of different age and of newborn infants; these proteins were absent from the blood serum of pregnant women and donors. They received the names of embryonic prealbumine 1, embryonic prealbumine 2 and embryonic alpha 2-globulin. The protein with the mobility of alpha 1-globulins was found in the amniotic fluid of foetuses and in the blood serum of pregnant women only and received the name of amniotic alpha 1-globulin. The concentration of the antigens in question was studied in the developing foetuses and in the blood serum of pregnant women at different stages of pregnancy.     

Immunochemical identification and physico-chemical characteristics of specific alpha 2 globulin of human granulocytes]

The antigen which has alpha 2-globulin mobility was identified immunochemically in leukolysate and pus extract. This antigen was localized in polymorphonuclear leukocytes by the immunofluorescence technique in the blood of healthy donors. The alpha 2-globulin of granulocytes (alpha 2GG) is not identical to lactoferrin, lysozyme, granulocyte elastase, fibronectin, fetal hemoglobin, amyloid P-component of the serum. The molecular mass of alpha 2GG is equal to 50 +/- 8 Kd. in the pus extract. The biological role of alpha 2GG is unknown.     

Immunochemical identification of human kidney-specific alpha2-macroglobulin]

Renospecific alpha2-macroglobulin (RS) with molecular weight 2 000 000 was identified by methods of immunochemical analysis. The revealed antigen was not identical to alpha2H-globulin (ferritin), to reno-pancreatic alpha2-globulin, uromucoid and alpha 2-macroglobulin of blood serum. The RS alpha2-macroglobulin content in the tumour tissue of the kidney decreased as compared to that in the normal kidney, in 16 of 23 tumors it was not revealed. On the sensitivity level of the monospecific RS alpha2-macroglobulin test-system it was not demonstrated in the blood of healthy persons and in the blood and urine of nephrologic patients.     

Immunochemical identification of beta2-globulin in metastases of ovarian tumors

Antisera were obtained upon immunization of rabbits with the extracts obtained from metastatic tissues of primary ovarian carcinoma into the omentum. Eight antigens were found, which were termed "ovarian-metastatic antigens" (OMA). OMA 1-7 showed cross-reactions with the antigens of one of the normal organs of the adult man: kidneys, spleen, brain. OMA-8 was not identified in the internal organ tissues of the adults and fetuses. Using immunodiffusion method, OMA-8 was revealed in the tissues of metastases of primary ovarian cancer into the omentum in 55% of cases (3-180 mg/l), in 50% of cases it was detected in primary ovarian carcinomas (3-50 mg/l) and in mature placenta (38-40 weeks) (6-12 mg/l). OMA-8 was detected in the chorion (8-20 weeks) and in the amniotic fluid at all periods at the maximum sensitivity of immunodiffusion method (1-2 mg/l). OMA-8 is a beta 2-globulin of protein nature with NW 35 kD, containing alpha- and beta-subunits with MW 18 and 19.5 kD. No carbohydrates, lipids and ferrum were determined in OMA-8. Its physico-chemical and antigenic properties differ from those of the described carcinoembryonic and placental proteins.     

Immunochemical characteristics of alpha 2-globulin from the rat "pregnancy zone" in ontogeny

An antigen with electrophoretic mobility of alpha 2-globulins was found in the blood serum of the pregnant rats by means of electrophoresis. It is detected not only in the blood serum and tissue extracts of the pregnant rats, but occurs in low concentrations (1 microgram/ml) in the blood serum of the normal adult animals. The concentration of this protein attains the maximal values by the end of the pregnancy period. Alpha 2-globulin appears to be a protein associated with pregnancy.     

Immunochemical detection, physicochemical characterization and levels of pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG) in seminal plasma of men

Pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG), the major secretory protein of the human uterine endometrium during the luteal phase of the menstrual cycle and early first trimester of pregnancy, has been detected by immunochemical techniques in seminal plasma. Biochemical analysis and immunoblotting has verified that immunoreactive alpha 2-PEG in seminal plasma exhibits properties identical to those of endometrial alpha 2-PEG, i.e. Concanavalin A-binding dimeric glycoprotein of native Mr 56,000, subunit Mr 28,000, average pI 4.7 and of alpha 2-mobility. Concentration of alpha 2-PEG in seminal plasma was 22.13 +/- 2.82 micrograms/ml (mean +/- s.e.m., n = 110) which compared to 12.02 +/- 1.65 micrograms/ml (mean +/- s.e.m., n = 48) found in amniotic fluid at 11-20 weeks of pregnancy, to 4.29 +/- 1.66 micrograms/ml (mean +/- s.e.m., n = 15) in uterine luminal fluid in women during the luteal phase and to 0.245 +/- 0.025 micrograms/ml (mean +/- s.e.m., n = 10) in sera at 10 weeks of pregnancy. This distribution is very different from that observed for pregnancy-associated placentally-derived serum proteins detected in seminal plasma. The source of alpha 2-PEG in seminal plasma is unknown but is unlikely to be the testis because of the normal levels observed in vasectomized men. In the endometrium alpha 2-PEG synthesis and secretion appears to be related to progesterone-dependent differentiation of the glandular epithelium. Therefore these observations suggest that a different mechanism of regulation of the gene for alpha 2-PEG operates in the male reproductive tract.     

Identification, isolation and physico-chemical properties of neurospecific alpha2-globulin]

In the immunization process of rabbits with the protein fraction of the water-salt extract from human brain partially and concentrated at 500-600 times was received antiserum revealed brain specific alpha 2-globulin that is not identical to the known cytoplasmatic brain specific antigens. This antigen has got electrophoretic mobility of alpha 2-globulins, molecular weight 90 +/- 10 kD and isoelectric point 4.1-5.4. Develop the procedure for purification of this antigen on the basis of the combination ion change, affinity, hydrophobic chromatography gel-filtration and isochromatofocusing.     

Gastric inhibitory polypeptide (GIP) concentration in human amniotic fluid

Several studies report that placenta and amniotic fluid (AF) may be a source of many peptide hormones. Although the presence of gastric inhibitory polypeptide (GIP) in amniotic fluid has not been described, it is present in the fetal gut. In this study we report the presence of insulin and GIP in human AF of normal and diabetic pregnancies. GIP concentrations in the AF collected two hours after an arginine tolerance test (ATT), at 34-36 weeks of gestation, were evaluated in 8 normal and 53 diabetic pregnant women. GIP was found in all samples of AF. The mean AF-GIP concentrations were 133 +/- 19 pmol/l in controls and 111 +/- 6 pmol/l in the diabetics, being the GIP values of the diabetics belonging to White Class B significantly lower than those of normals (99 +/- 10 vs 133 +/- 19 pmol/l). The GIP/IRI molar ratio was significantly lower in the diabetics than in controls (1.2 +/- 0.2 vs 2.5 +/- 0.4); moreover the GIP/IRI molar ratio was significantly higher in AF collected from diabetic pregnant women who delivered overweight infants than in AF of normal weight infants or controls. This finding would suggest a negative feedback mechanism between GIP and insulin in fetus.     

Cholinesterases (acetyl and pseudocholinesterase) in human amniotic fluid]

Study on 41 cases. The cholinesterasic activity of human amniotic fluid, free from blood, is very weak, in average 0,10 microkatal, approximatively 300 times weaker than mother's serum. In this "total cholinesterase", we find a large amount (about 40%) of acetylcholinesterase. Amniotic cholinesterases have an prominent placentary origin, with, in addition, a likely activity proceeding from cellular elements.     

Comparative immunochemical and physicochemical characteristics of human placental chorionic alphal- and alpha2-microglobulins]

Immunochemical analysis showed chorionic alpha1-microglobulin to be immunologically different from chorionic alpha2-microglobulin. Some physico-chemical properties of these proteins were studied, and they were found to differ from one another by a number of parameters.     

Detection of stage specific and organ specific human brain antigens during embryogenesis]

The rabbit antisera were obtained against the water soluble antigens of the brain of 8--10 weeks old human foetuses. Three groups of specific antigens were identified in the brain of human foetuses: 1) antigens common for the embryonic brain and other organs of the same age; 2) antigens common for the embryonic brain and some organs of the adult organism; 3) stage (phase)-specific brain antigens present only in the brain between the 8th and 10th weeks of pregnancy.     

Immunochemical comparison of human and goat haptoglobin and alpha 2-macroglobulin.

1. A non-haptoglobin alpha 2-globulin was observed in the plasma of goats (Capra hircus) after inflammatory stresses. 2. This stress reaction protein exhibited an electrophoretic mobility and molecular weight similar to goat haptoglobin. 3. Complete immunological cross-reactivity was demonstrated between the goat protein and human alpha 2-macroglobulin. 4. The functional and evolutionary significance of these observations are discussed.     

Identification, purification and physico-chemical properties of neurospecific alpha 1-globulin]

The partially purified and concentrated 500-600-folds protein fraction has been obtained from human brain extract. This protein fraction was used for the immunization of rabbits. The corresponding anti-sera have the potency to detect the brain specific alpha 1-globulin, which is not identical to known cytoplasmatic brain specific protein. These antisera were used for the control of the antigen purification procedure which included ion-exchange, affinity and hydropho'ic chromatography, gel-filtration and isochromatofocusing. The antigen, purified to the homogeneity, has the electrophoretic mobility of the alpha 1-globulins, M(r) = 110-10 kD, and isoelectric point at pH 2.9-3.1.