[Effects of complexes of alpha-fetoprotein and amphiphilic docosaenoic acid derivatives on the humoral immune response in C57BL/6 mice]. / Vliianie kompleksov al'fa-fetoproteina s amfifil'nymi soedineniiami na osnove dokozageksaenovoi kisloty na gumoral'nyi immunnyi otvet u myshei linii C57BL/6.

The immunotropic activity of three derivatives of docosahexaenic acid (N-docosahexaenoyl-L-serine phosphate, N-docosahexaenoyl-L-threonine phosphate, and N-docosahexaenoyl-L-tyrosine phosphate) in complexes with high-purity human alpha-fetoprotein was evaluated. The polyene compounds stimulate the humoral immunity in mice immunized with T-dependent antigen (ram red blood cells). The immunotropic activity of the alpha-fetoprotein-ligand complexes studied depends on the structure and the content of polyene ligands.

[Autoantibodies to human thyroid peroxidase in immunoassay and immunoaffinity chromatography]. / Autoantitela k tiroperoksidaze cheloveka v immunoanalize i immunoaffinnoi khromatografii.

A biochemical system of radioimmunoassay of human thyroid peroxidase (TPO) with the use of specific autoantibodies was developed for the first time. This system includes lyophilized preparations of human autoantibodies to TPO, radioiodinated TPO, standards prepared from pure TPO, and the solid-phase protein A as a precipitating agent. An analytical system was used to study the TPO distribution in fractions of thyroid tissue homogenate. Differences in TPO concentrations in mitochondria, microsomes, and supernatant fluid depending on thyroid pathology and tissue storage conditions were found. The behavior of immobilized anti-TPO-autoantibodies in immunoaffinity chromatography of this microsomal antigen was studied.

Modification of humoral immune response in C57Bl/6 mice with a complex of alpha-fetoprotein and retinoid acid derivatives.

Amides of all trans-retinoic acid and O-phospho-L-threonine, O-phospho-L-tyrosine, and O-phosphoethanolamine injected intravenously in a dose of 6.8 microg/kg 1.5-3.4-fold increased the count of antibody-producing cells in the spleen of C57Bl/6 mice (primary immune response to sheep erythrocytes). Activity of the complex containing alpha-fetoprotein and L-threonine ligand did not differ from that of free retinoid. This complex holds much promise as a prototype of immunomodulators with a wide range of activity.

[Interaction of transcortin with human syncytiotrophoblast]. / Vzaimodeistvie transkortina s sintsitiotrofoblastom cheloveka.

Interactions of transcortin (corticosteroid-binding globulin, CBG) variants, nCBG and rCBG, present in the blood of pregnant women, and microvesicular fraction of the brush border membrane of human placental syncytiotrophoblast at 23 +/- 2 degrees C were studied. Interaction of nCBG in complex with a steroid with each of the two types for specific binding was found associated with transmembranous transfer of glycoprotein. Interaction of rCBG with binding sites of both types did not involve subsequent glycoprotein transfer through the membrane. Possibility of penetration of only one CBG variant through syncytiotrophoblast membrane suggests the presence of different mechanisms of these glycoproteins' participation in the hormonal effects of steroids associated with them.

[The effect of steroids on binding of transcortin with human syncytiotrophoblasts]. / Vliiane steroidov na sviazyvanie transkortina so sintsitiotrofoblastom cheloveka.

The effects of steroids on specific binding of transcortin (corticosteroid-binding globulin, CBG) variants (nCBG and vCBG) from pregnant women sera to the microvesicular membrane fraction derived from placental syncytiotrophoblast brush border have been studied. It was shown that native CBG variants stripped of steroids lose their ability to interact with specific membrane sites. Specific binding of CBG variants was found to increase in parallel with an increasing per cent content of glycoproteins associated with cortisol, reaching a maximum at cortisol concentrations of 5.10(-8)-5.10(-7) M which is saturating for the complex formation. Specific binding sites in the syncytiotrophoblast membrane do not differentiate between the nCBG complexed to cortisol, corticosterone, progesterone or testosterone but show a selectivity towards the vCBG-cortisol complex.

[Isolation, identification and properties of a new thyroxine-binding protein--apolipoprotein A-1 from human blood serum]. / Vydelenie, identifikatsiia i svoistva novogo tiroksinsviazyvaiushchegobelka--apolipoproteina A-1 syvorotki krovi cheloveka.

A protein having a molecular mass of about 25 kWa was isolated by thyroxin (T4)-Sepharose affinity chromatography from human blood serum; its properties were found to be distinct from those of known T4-binding proteins. Using immunodiffusion, radioimmunoassay, lipid analysis, differential precipitation and electrophoresis, it was shown that the isolated protein is a component of high density lipoprotein (HDL) particles and represents an apolipoprotein A-1 (apoA-1). Using cholate-Sepharose chromatography apoA-1 was separated from the lipid moiety and contaminant proteins, and apoA-1 was effectively isolated directly from the blood serum. Apo-A-1-HDL and apoA-1 obtained by affinity chromatography as well as the HDL3 fraction isolated by a standard ultracentrifugation technique, all displayed a T4-binding activity, the affinities for the hormones being of the same order of magnitude. The T4 interaction with these preparations induced difference UV-absorption signals, altered the characteristics of apoA-1 intrinsic fluorescence without affecting the circular dichroism of the protein-hormone system. The binding of spin-labelled T4 to apo-1, apoA-1-HDL or HDI3 caused substantial changes in the shape of the ESR spectrum and an increase in the apparent rotational correlation time. The mobility of the radical fragment of spin-labelled T4 depended on the composition and properties of the protein preparation. The electron spectroscopy data suggest that the T4-HDL interaction occurs via specific mechanisms and that the molecular structures of the complexes formed thereby are not characteristic of other known T4-binding proteins.

[Pregnancy-associated molecular variation of thyroxine-binding globulin in health and in various diseases of man]. / Sviazannaia s beremennost'iu molekuliarnaia raznovidnost' tiroksinsviazyvaiushchego globulina v norme i pri nekotorykh zabolevaniiakh u cheloveka.

A molecular variant of human serum thyroxine-binding globulin (TBG), containing only triantennary oligosaccharide chains and having a more prolonged in vivo survival (TBG-1), was first detected in normal pregnancy and then in the postpartum period. Serum TBG-1 was measured in normal and in some pathological conditions associated with normal or increased TBG biosynthesis using a combination of Con A-Sepharose 4B microcolumn affinity chromatography and a highly sensitive TBG radioimmunoassay. TBG-1 was shown to be present in the sera of healthy subjects (0.21 +/- 0.04 micrograms/ml, n = 60; 1.15% of total TBG). The proportion of TBG-1 in total serum TBG was significantly increased (up to 9.5%) in conditions with serum TBG excess (cancer, hypothyroidism and liver diseases). It has been assumed that a selective enhancement of biosynthesis of the TBG molecular variant with a prolonged half-life in the circulation during the posttranslational modification of the polypeptide chain is a response to the body requirements in a high TBG concentration.

[Specific binding of iodothyronines with apolipoprotein A-1 in high density lipoprotein particle isolated from human serum by affinity chromatography on thyroxine-sepharose]. / Spetsificheskoe sviazyvanie iodtironinov s apolipoproteinom A-1 v sostave lipoproteidnykh chastits vysokoi plotnosti, vydelennykh iz syvorotki cheloveka affinnoi khromatografiei na tiroksin-sefaroze.

The kinetic and equilibrium characteristics of interaction of thyroxine (T4) and its structural analogs with a high density lipoprotein (HDL) fraction isolated from human serum by T4-Sepharose affinity chromatography and containing apolipoprotein A-I (apo A-I) as a sole protein component, were studied. The binding of [125I]T4 to apo A-I-HDL reached a maximum after 40 min and did not change during the next 80 min of incubation at 0 degrees--22 degrees C. Dissociation of [125I]T4 induced by the addition of excess unlabeled T4 to the complex solution proceeded more intensely on a time scale at 0--2 degrees C than at 22 degrees C. Incubation of apo A-I-HDL with increasing concentrations of T4 showed that the binding is saturable. The data analysis using different computer programs revealed the presence in apo A-I-HDL of a single class of binding sites with K alpha = (4.0 +/- 2.1).10(-7) M- and Bmax = 1.7 +/- 0.8 nmol T4/mg of protein. Naturally occurring iodothyronines, their analogs and D-isomers of thyroid hormones competed with [125I]T4 for the binding sites on apo A-I-HDL with the following inhibitory potencies: L-T4 = D-T4 greater than or equal to 3,3',5-triiodo-L-thyronine = 3,3',5-triiodo-D-thyronine greater than 3,5-diiodo-L-thyronine = 3,3',5- triiodothyroacetic acid greater than 3,3',5-triiodothyropropionic acid greater than or equal to 3,5-diiodo-L-thyrosine.(ABSTRACT TRUNCATED AT 250 WORDS)

[The role of steroid-binding glycoproteins in human blood in transmitting the hormonal signal to the cell]. / Rol' steroidsviazyvaiushchikh glikoproteinov krovi cheloveka v peredache gormonal'nogo signala k kletke.

The data on interaction of specific steroid-binding glycoproteins of blood plasma and of their steroid-hormone complexes with the plasma membranes of various hormone target tissues suggests that these glycoproteins play an active role in the transport of steroid hormones into the target cells. Both polypeptide and carbohydrate components of the steroid-binding glycoproteins are involved in the formation of determinants for the membrane recognition of hormone-glycoprotein complexes. This causes a modulation of the steroid hormone action on various tissues due to modifications of the glycoprotein carbohydrate structures.

[Affinity chromatography of thyroxine-binding proteins from human serum]. / Affinnaia khromatografiia tiroksinsviazyvaiushchikh belkov syvorotki krovi cheloveka.

The affinity matrix prepared by the attachment of L-thyroxine (T4) to epichlorohydrine-activated Sepharose 4B biospecifically absorbs the T4-binding globulin (TBG), 25K and 80/27K proteins, immunoglobulin G (IgG) and albumin (HSA) from human normal and retroplacental sera. The absorbed protein patterns were shown to depend on the immobilized T4 concentration, pH, temperature and incubation time. The potent eluents desorbing 85-100% of the protein are 1 mM NaOH, 3 M NH4SCN, 10(-5) M T4 or 3 mM 8-anilinonaphthalene-1-sulfonic acid (ANS) for TBG; NaOH, NH4SCN, 3 mM MgCl2 or 12mM sodium cholate for 25K protein and HSA; NaOH, NH4SCN or MgCl2 for the 80/27K and 25K proteins and IgG. Moreover, T4 desorbs small amounts (6-8%) of the 80/27K and 25K proteins, while sodium cholate elutes about 6% of TBG. The eluted from T4-Sepharose 4B and further purified TBG, 25K and 80/27K proteins display different [125I]T4-binding activities within the pH range from 2 to 9 and differ by their resistance to thermal inactivation at 50-80 degrees C. Double radial immunodiffusion analysis with the use of antisera to TBG, 25K, 80/27K, HSA and IgG demonstrated that the proteins share no common antigenic determinants. It was concluded that the novel 25K and 80/27K proteins represent endogenous components of the human blood thyroid hormone-binding protein system rather than fragments or aggregates of the known T4-binding proteins.

[Experience with the use of laboratory methods of comprehensively evaluating the state of health of newborn infants]. / Opyt ispol'zovaniia laboratornykh metodov kompleksnoi otsenki sostoianiia zdorov'ia novorozhdennykh.

The authors offer new laboratory methods for comprehensive assessment of the health status of the newborns. Introduction of these methods in practical activity of clinical diagnostic laboratories of therapeutic and prophylactic institutions will promote early diagnosis of some diseases in the newborns and help comprehensively assess the neonates' adaptation potential, this being valuable for successful management of the babies.

[Characteristics of a transcortin-binding component of the plasma membrane of cells of the human decidual endometrium]. / Kharakteristika transkortin-sviazyvaiushchego komponenta plazmaticheskoi membrany kletok detsidual'nogo éndometriia cheloveka.

A sialoglycoprotein was isolated by affinity chromatography on immobilized transcortin from plasma membranes of human decidual endometrium cells, whose components were labeled with 125I and solubilized with sodium cholate. The apparent molecular mass of the monomer is 20.0 +/- 1.5 kDa, pI is at pH 3.3. The sialoglycoprotein specifically binds transcortin complexed to progesterone with Kd approximately 10(-10) M.

[New aspects of isolation and purification of thyroxine-binding globulin from human biological fluids]. / Novye aspekty vydeleniia i ochistki tiroksinsviazyvaiushchego globulina iz biologicheskikh zhidkostei cheloveka.

Based on the new data concerning the multicomponent system of thyroxine-binding proteins in human plasma, some methodological aspects of isolation and purification of thyroxine-binding globulin (TBG) were examined, and a simple two-step procedure for TBG purification was developed. Normal human blood serum, retroplacental serum and amniotic fluid were used as TBG sources. The procedure includes affinity chromatography and adsorption chromatography on a hydroxyapatite column. A biospecific adsorbent was synthesized by stepwise binding of epichlorohydrin and thyroxine to Sepharose. The yield of pure TBG varied from 60 to 80%, depending on the TBG source used. The properties of TBG preparations from retroplacental serum and amniotic fluid were identical; both preparations contained a pregnancy-associated molecular variant, TBG-1. Two novel serum thyroxine-binding proteins were detected, isolated and partly characterized.

[Biological function of the carbohydrate component of human sex hormone-binding globulin]. / O biologicheskoi funktsii uglevodnogo komponenta sekssteroidsviazyvaiushchego globulina cheloveka.

The role of the carbohydrate component of sex steroid-binding globulin (SBP) from human blood in the glycoprotein interaction with the recognition system for SBP-estrogen complexes in human decidual endometrium plasma membrane was studied. It was shown that the removal of N-acetylneuraminic acid residues from the oligosaccharide chains of SBP did not affect the steroid-binding or immunochemical properties of the glycoprotein. At the same time, the above modification of the glycoprotein resulted in a loss by SBP of its ability to specifically interact with the membrane recognition system. It is concluded that the oligosaccharide chains of SBP are involved in the formation of determinants needed for recognition of the SBP-estrogen complexes by endometrium cell plasma membranes.

[Interaction of the transcortin-progesterone complex with plasma membranes of human decidual epithelium]. / Vzaimodeistvie kompleksa transkortin-progesteron s plazmaticheskimi membranami kletok detsidual'nogo éndometriia cheloveka.

It was demonstrated that transcortin complexed with progesterone specifically binds to plasma membranes of human decidual endometrium, a progesterone target tissue. This interaction is characterized by a high affinity [Kd = (1.0 +/- 0.2).10(-10) mol/l] and selectivity. Such human serum proteins as albumin, orosomucoid, transferrin, thyroxine- and sex steroid-binding globulins do not compete with transcortin for the binding sites on the membranes. The concentration of endogenous transcortin in sodium cholate-solubilized endometrium cell membranes was determined by the radioimmunoassay method.

[Radioimmunologic system for the quantitative determination of thyroxine in human blood serum]. / Radioimmunologicheskaia sistema dlia kolichestvennogo opredeleniia tiroksina v syvorotke krovi cheloveka.

A methodological approach to development of a radioimmunological system for quantitative assay of thyroxine (T4) in human blood serum is described. It implies investigation of every component of the system and matching of the components by the physico-chemical properties and concentrations. An antiserum to the T4 conjugate was prepared with bovine serum albumin. The hapten "fitting density" was 30 mol per 1 mol of protein. The antiserum had a titer of 5000, was highly specific and showed affinity to T4, Ka (7.6 +/- 1.0) X 10(8) M-1. The characteristics of the antiserum did not depend on concentration of the hydrogen ions in solution at pH 6.0-9.0 and the ionic strength of the buffer solutions within M 0.05-1.0. 8-Aniline-1-naphthalene sulfonic acid (ANA) in concentrations of 0.05-0.1 per cent completely inhibited binding of T4 to endogenic proteins of the blood serum and had no effect on the antiserum interaction with T4. This provided performance of the radioimmunological assays at 0.1 per cent concentration of ANA. [125I]T4 with the specific activity of 40 TBk/g was used in the study. The quality of [125I] T4 was estimated by its immune reactivity. The standard solutions of T4 were prepared with donor blood serum as the basis. It was shown that quantitative assays of T4 in blood serum within clinically significant ranges of the hormone concentration are possible at 1:5000 final dilution of the antiserum, the [125I] T4 activity of 20 000-30 000 imp/min per a specimen and 5 per cent content of the blood serum in the analytical specimen. A simple and reliable technique for T4 assay in human blood serum is developed.

[Comparative characteristics of molecular forms of human thyroxine-binding globulin]. / Sravnitel'naia kharakteristika molekuliarnykh raznovidnostei tiroksinsviazyvaiushchego globulina cheloveka.

Two molecular variants, of thyroxin-binding globulin (TBG), TBG-1 and TBG-2, were obtained from human retroplacental blood by fractionation of pure TBG on concanavalin A-Sepharose. It was found that both variants are immunologically identical, have similar molecular weights, amino acid composition and spectral properties, and possess the same affinity for thyroid hormones. However, TBG-1 and TBG-2 differed in charge upon isoelectrofocusing and had different monosaccharide composition. The existence of two molecular variants of TBG in pregnancy is probably due to the peculiarities of the polypeptide chain glycosylation during TBG biosynthesis.

[Radioimmunological system for the quantitative determination of progesterone in human blood serum]. / Radioimmunologicheskaia sistema dlia kolichestvennogo opredeleniia progesterona v syvorotke krovi cheloveka.

A methodical approach to development of a radioimmune system for quantitative assay of progesterone in the blood serum of man is described. It implies investigation of every component in the system and conformity of these components by the physico-chemical properties and concentrations. An antiserum to the progesterone conjugate with bovine serum albumin was prepared. The "fitting density" of the heptaene was 35 mol/mol of the protein. The linkage with the carrier protein in it was performed by the functional group at C(ii) of the steroid molecule. The antiserum had a high titre (28000), high specificity and affinity to progesterone (Ka (3.5 +/- 0.4) X 10(9) M-1). The antiserum characteristics did not depend on the hydrogen ion concentration in the solution at pH 3.5-10.0 and the ionic strength of the buffer solutions (0.05 to 1.0 M). This allowed performing the radioimmunological assay at pH 3.5. Under such conditions complete inhibition of the endogenic binding proteins of the blood serum was possible. 125I preparations of progesterone made in the USSR were used in the study. The progesterones were prepared by adding the molecule of histamine containing the 125I atom to the molecule of progesterone modified at C(3). The quality of the 125I preparations of progesterone was estimated by their immune reactivity. The standard solutions of progesterone were prepared with the donor blood serum. It was shown that quantitative determination of progesterone in the blood serum at clinically significant ranges of the hormone concentration was possible at the final antiserum dilution of 1:28000, the label activity of 20000-30000 imp/min per a sample and the content of the blood serum in the analyzed sample equal to at least 10 per cent. A simple and reliable procedure for the assay of progesterone in the blood serum of man was developed.

[Interaction of blood sex steroid-binding globulin-steroid complexes with plasma membranes of human decidual endometrium cells]. / Vzaimodeistvie kompleksov sekssteroidsviazyvaiushchii globulin krovi-steroid s plazmaticheskimi membranami kletok detsidual'nogo éndometriia cheloveka.

Plasma membranes of decidual tissue cells specifically bind the sex steroid-binding globulin (SBP) complexes with estrogen (estradiol, estriol, estrone) and with the pharmacological agent danazol but do not interact with the SBP-testosterone or SBP-dihydrotestosterone complexes. The selectivity of interaction of the SBP-steroid complexes with decidual tissue cellular membranes provide evidence for the active role of SBP in the realization of steroid effects on the target tissue.