IgG4 antibodies in hay fever patients. Difference in IgG4 response to tree pollen and grass pollen allergens.

Total IgE and IgE and IgG4 antibodies to offending allergens were studied in 118 hay fever patients allergic to tree pollen and grass pollen allergens. No difference in the level of total IgE and IgE antibodies was found between these two groups while the IgG4 antibody level was significantly higher in tree pollen allergic patients. It is suggested that the IgG4 antibodies to tree pollen allergens can be stimulated by structurally related allergens of vegetable food and are probably involved in tree pollen-associated food hypersensitivity.

[alpha 1-proteinase inhibitor in breast cancer]. / alfa1-Ingibitor proteinaz pri rake molochnoi zhelezy.

Level of alpha 1-proteinase inhibitor (alpha 1-Pi) and antitryptic activity in blood serum were assessed in 167 patients with breast cancer and 30 cases of benign lesions. An increase in blood serum-alpha 1-Pi level was shown to be associated with tumor advancement but could not be regarded as a marker of cancer. Concentration of the inhibitor exceeded 8 mg/ml in 83.4% of cases with metastatic breast cancer but was as a rule, equal to or below that value in metastasis--free patients. Those two groups were prognostically different. Morphological analysis established a correlation between blood serum-alpha 1-Pi level, on the one hand, and grade of malignancy, degree of pathologic changes in tumor stroma and extent of lymph node involvement, on the other.

[IgG4 antibodies to the allergens of grass and tree pollens in pollinosis patients: their diagnostic and prognostic significance]. / IgG4-antitela k allergenam pyl'tsy zlakov i pyl'tsy derev'ev u bol'nykh pollinozom: diagnosticheskoe i prognosticheskoe znachenie.

Total IgE, IgE and IgG4 antibodies to offending allergens were studied in 118 hay fever patients with allergy to tree pollen or to grass pollen. No difference in the level of total IgE and IgE antibodies was found between these groups of patients while the IgG4 antibody level was significantly higher in tree pollen-allergic patients. It has been established that in tree pollen allergy, the determination of IgG4 antibodies is of no less diagnostic value than the assay of IgE antibodies. The authors discuss the problems of the prognostic value of IgG4 antibody increase during the hyposensitization therapy and of a possible relationship between IgG4 antibodies and allergy to vegetable food often associated with tree pollen allergy.

[Immunoenzyme test system with monoclonal antibodies to human IgG4 in the determination of allergen-specific antibodies in pollinosis]. / Immunofermentnaia test-sistema s monoklonal'nymi antitelami k IgG4 cheloveka dlia vyiavleniia allergenspetsificheskikh antitel pri pollinoze.

ELISA for determination of allergen-specific IgG4 antibodies was developed with the help of monoclonal anti-IgG4 antibodies obtained by classic hybridoma technique. Subclass specificity of antibodies were studied in sera of 108 patients suffering from pollinosis. Antibodies of this isotype were found in the majority of patients with tree pollen allergy but not in patients with grass pollen allergy. The level of IgG4 antibodies correlated with the severity of the disease but not with the intensity of skin tests. Specific hyposensitization resulted in significant increase of IgG4 antibody level in patients with tree pollen allergy. Determination of IgG4 antibodies is proved to be useful to reveal tree pollen allergy and to monitor hyposensitization therapy.

[Purification of human alpha-1-protease inhibitor and production of antisera for its immunochemical determination]. / Ochistka alpha-1-ingibitora proteaz cheloveka i poluchenie antisyvorotki, prigodnoi dlia ego immunokhimicheskogo opredeleniia.

Human homogeneous alpha-I-inhibitor of proteases was isolated and purified by means of chromatography on DEAE cellulose, hydroxyapatite and concanavalin Sepharose 4B. After immunization of rabbits specific monovalent antiserum towards the inhibitor was produced. The antiserum was used in diagnosis of hereditary deficiency of alpha-I-inhibitor of proteases and for estimation of its content in newborns.

[Alpha 1-protease inhibitor: characteristics of its biochemical and biological properties and its level in various diseases (review of the literature and personal observations)]. / Alpha 1-ingibitor proteaz: kharakteristika biokhimicheskikh i biologicheskikh svoistv i opredelenie urovnia pri razlichnykh zabolevaniiakh (obzor literatury, sobstvennye nabliudeniia).

The authors review biological and biochemical properties and the clinical importance of the serum proteases alpha 1-inhibitor with broad-range antiproteolytic activity. Congenital deficiency of this protein is a frequent enough condition linked with predisposition to some diseases of the lungs and liver. Early determination of the deficiency of alpha 1-Pi is fairly urgent, since it permits the early administration of the preventive measures and substitution therapy. The immunochemical technique makes it possible to determine all the varieties of alpha 1-Pi, as they are antigenically similar. Sera from 267 patients with different diseases were examined. The content of alpha 1-Pi was found to be elevated in Waldenström's macroglobulinemia, chronic active hepatitis and liver cirrhosis and to be lowered in bronchial asthma. In multiple myeloma and pollinoses, no alterations in the alpha 1-Pi content were recorded.

[Complexes of monoclonal immunoglobulins of various classes with albumin and alpha 1-proteinase inhibitor]. / Kompleksirovanie monoklonal'nykh immunoglobulinov raznykh klassov s al'buminom i alpha 1-ingibitorom proteaz.

Ability of monoclonal immunoglobulins (IgG, IgA, IgM and IgD) to develop complexes with albumin and alpha 1-proteinase inhibitor (alpha 1-PI) was studied using methods of immunoelectrophoresis, cross immunoelectrophoresis and immunoselection. High of proteins, producing complexes with albumin, was found among monoclonal IgA, IgM and IgG; alpha 1-PI formed complexes mainly with IgA and IgM. In healthy volunteers complexes of blood serum proteins were not found. Albumin and alpha 1-PI developed complexes most often with various molecules of monoclonal immunoglobulins. Ability to produce complexes did not depend on the type of the paraprotein.

Crystal-forming proteins of Bacillus thuringiensis. The limited hydrolysis by endogeneous proteinases as a cause of their apparent multiplicity.

The crystals of the entomocidal protein of Bacillus thuringiensis are admixed with proteinases that in the course of their dissolution cause gradual degradation of the "genuine" crystal-forming protein components (i.e. the primary biosynthetic products) to products of lower molecular weight. This phenomenon might explain at least partially the contradictory data on the molecular parameters of the crystal-forming proteins. Preliminary inactivation of the proteinases adsorbed on the crystals allowed us to eliminate this source of the artefacts and to gain more reliable data on the protein composition of the crystals formed by various strains of B. thuringiensis. It has been shown that the crystals formed by all serotypes of B. thuringiensis, with the exception of the serotype V, contain only one protein with a mol. wt. of 145000, 135000 or 130000, depending on the strain. The majority of the strains that belong to the serotype V form crystals consisting of two proteins with mol. wts. of 135000 and 130000, but some of them also have a third component with a mol. wt. of 65000.

[Using a combined transcription-translation system for determining the role of cryptic plasmids of Bacillus thuringiensis]. / Ispol'zovanie sovmeshchenoi sistemy transkriptsii--transliatsii dlia vyiusneniia roli kripticheskikh plazmid Bacillus thuringiensis.

The possibility of entomocyde crystal protein synthesis was studied using a heterological cell-free system with Bacillus thuringiensis plasmid DNA as template. The high level of template activity is usual for Bac. thuringiensis plasmid DNA. Immunochemical studies of the in vitro synthesized polypeptides showed that Bac. thuringiensis plasmid DNA does not direct crystal protein synthesis.

[Proteinases in growth and spore formation of Bacillus thuringiensis]. / Proteinazy v protsesse rosta i sporoobrazovaniia Bacillus thuringiensis.

Some serine proteases and leucine aminopeptidases were detected inside and outside the cells during the analysis of three crystalline and two acrystalline strains of Bac. thuringiensis var. galleriae. The data obtained on the protease formation during growth and sporulation and the level of their activity are indicative of intracellular proteases involvement in spore- and crystal formation. The enzymes isolated from the culture medium do not probably take part in these processes. The intracellular enzymes may account for the different crystal protein composition of various strains due to limited proteolysis of crystal proteins in the course of biosynthesis.

[Proteolytic enzymes bound to Bac. thuringiensis crystals]. / Proteinazy, Sviazannye s kristallami Bac. thuringiensis.

It was demonstrated that crystals of entomopathogenic protein from Bac. thuringiensis contain admixture of proteinase either adhered to their surface on inconponated into crystal lattice defects. A proteolytic action, particularly when enhanced by crystal dissolution, causes progressive degradation of crystal proteins with molecular weights of 140 000--129 000 down to the components with smaller molecular weights. This may, at least, partially account for the contradictions in the literature data on crystal composition. Using synthetic peptide substrates and specific inhibitors, it was shown that the enzymes incorporated into crystals belong to serine and metalloproteases. The presence of leucine aminopeptidase was also noted. A method for enzyme separation from crystal has been developed.

[Proteins of Bacillus thuringiensis delta-endotoxin crystals]. / Belki kristallov delta-endotoksina Bac. thuringiensis.

Pure crystals (at least 99% purification) of sigma-endotoxin were isolated from Bac. thuringiensis var. galleriae. The complete dissolution of crystals might be achieved by the increase of pH up to 12 and higher or by a combined action of S = S-reducing and denaturing agents. Electrophoresis of the solubilized crystal proteins in 5% polyacrylamide gels containing 0,1% sodium dodecyl sulfate and 8 M urea reveals two major bands corresponding to molecular weights of 120000--140000 (65%) and 65000 (8-10%), and some minor components whose molecular weights varied from 65000 to 340000. Urea (3--8 M) causes to partial dissolution of the crystals; the component with molecular weight of 65000 is mainly found in the solution (component A). In dithioerythritol extracts at pH 9 the major component of the crystal is the protein with molecular weight 120000--140000 (component B). The crystals, alkali-soluble components and proteins isolated from crystals by selective extraction (3--8 M urea or 0.01 M dithioerythrytol, pH 9) were found toxic for the larvae of Galleria mellonella.