Use of immunoenzyme analysis for determining antibacterial antibodies in diphtheria infection

A diagnostic EIA system for the detection of antibacterial antibodies in diphtheria infection has been developed. As antigen, homogeneous membrane protein (mol. wt. 64 KD) obtained from Corynebacterium diphtheriae cell walls has been used. This protein antigen has been prepared with the use of nonionic detergent NP-40.     

Specific antibodies and their role in the formation of antidiphtheria immunity

In human sera, studied with the use of the enzyme immunoassay, antidiphtheria postvaccinal antitoxic IgG and naturally acquired antibacterial IgG, IgM and IgA were detected. In the blood of children and adults aged up to 50 years antitoxic IgG were present at normal and high concentrations. In 50% of children antibacterial IgA were absent, while specific antibacterial IgM could be found at high concentrations. Changes in the content of antibacterial antibodies of different classes in sera were observed with age. More than 90% of adults had antibacterial IgA and IgG at normal and hig concentrations, while the level of IgM decreased. Under the influence of ecological, social, anthropogenic and other environmental factors the optimum levels of specific antibodies were replaced by anomalous ones, which led to an increased number of persons susceptible to diphtheria infection and in the intensity of the circulation of the infective agent. The deficiency of antidiphtheria antibacterial antibodies in the blood determined the necessity of correcting immunity by means of not only toxoid, but also bacterial antigens.     

Immobilization of invertase in polyvinyl alcohol coatings

A possibility of invertase immobilization in the polyvinyl alcohol coating formed directly on the electrode surface from water solution of polyvinyl alcohol and boric acid was being investigated. Conditions for obtaining the polymeric coating at the constant potential and at the constant current were compared. In order to obtain the polymeric coatings with a marked enzyme activity optimal conditions were found.     

Structural features of the submandibular salivary gland of gnotobiotic rats

The submandibular salivary gland was histochemically and electron microscopically studied in gnotobiotic and conventional Wistar rats at the age of 15 days--10 months. By the first month of age, the submandibular salivary glands in both groups of animals complete differentiation of their acini, and the gland, according to the type of its secretion, becomes seromucous with predominance of albuminous component in it. Succinate dehydrogenase activity and nonspecific esterase predominate in epithelium of the excretory ducts. At the second month of life some signs of moreasing ductal part of the gland appear in the gnotobiotic rats. This part greatly increases by the 4th--10th month of life, mainly at the expense of twisted granular sections. This increase in number and volume of the twisted granular ducts results in compressing acini and in ultrastructural disorders of granulocytes, with destruction of some mitochondria, increasing number of lysosomes, decreasing size and alterations in character of secretory granules, in structural disorders of the laminar complex. All these manifest depressing secretory function of granulocytes in the gland of gnotobiotic animals.     

Humoral response to the injection of acellular Pertussis vaccine

The humoral response of mice and rabbits to the injection of whole-cell pertussis vaccine (PV) and acellular pertussis vaccine (APV), developed at the Mechnikov Research Institute for Vaccines and Sera (Russian Acad. Med. Sci.) in Moscow, was studied. In the sera of immunized animals antibodies to the antigenic complex were determined in the direct hemagglutination (DHA) test, specific antibodies to filamentous hemagglutinin (FHA) and pertussis toxin (PT)--in the enzyme immunoassay (EIA) and antibodies neutralizing PT in a cytopathogenic dose (CPD)--in neutralization test on Chinese hamster ovary (CHO) cells. In mice and rabbits immunized with APV the antibody titers determined in the DHA test were higher than those in the animals immunized with PV. Specific antibodies titers to FHA and PT in the sera of rabbits immunized with APV were also higher than those in the sera of rabbits immunized with PV. High dilutions of sera taken from the animals immunized with APC neutralized 4-16 doses of PT in the neutralization test on CHO cells. The most important result of this study was the detection of a more pronounced immune response in the animals immunized with APV in comparison with that induced by PV according to the results obtained in EIA and in the test of PT CPD neutralization on CHO cells.     

Immunomodulating properties of the antigens of the cell walls of nontoxigenic Corynebacterium diphtheriae

The total protein fraction was obtained by extraction with nonionic detergent NP-40. The fraction composition and molecular parameters were studied with DDS-Na electrophoresis in polyacryl amide gel. Common properties of the Corynebacterium diphtheriae protein fraction 66 kD and an analogous fraction of the cell walls of staphylococci and the measles virus were shown with immunoblotting. Cell-mediated mechanisms of the immune response to contact with antigens of Corynebacterium diphtheriae cell walls were revealed.