[Blood sampling systems and interference in immunoassay].

Immunoassay interference caused by reagents (additives, excipients) incorporated in the blood sampling systems is undetectable by the conventional methods for intralaboratory quality control, by using reference substances. A large number of manufacturers and a great physicochemical variety of additives in the current vacuum and non-vacuum systems require that additional measures be made to monitor systemic errors in immunochemical studies. The review considers the mechanisms of this type of interference and the problems of its monitoring and prevention.

[Effect of immunomodulators of various origin on the histamine-sensitizing activity of whole-cell pertussis vaccine]. / Vliianie immunomoduliatorov razlichnogo proiskhozhdeniia na gistaminsensibiliziruiushchuiu aktivnost' korpuskuliarnoi kokliushnoi vaktsiny.

Changes in the histamine-sensitizing activity of whole-cell pertussis vaccine (PV) under the action of immunomodulators (IM) of bacterial (peptide and peptidoglycans), synthetic (peptidoglycan) and vegetable origin have been studied. The study has revealed that these IM, introduced orally and parenterally, exhibit histamine-sensitizing activity, depending on the nature of IM and the optimum selection of the doses of IM and PV.

[The use of microdot immunoenzyme analysis with visual detection for the determination of tularemia antibodies]. / Ispol'zovanie mikrotochechnogo immunofermentnogo analiza s vizual'noi indikatsiei dlia opredeleniia tuliaremiinykh antitel.

The possibility of using the micropoint enzyme immunoassay (EIA) on a nitrocellulose membrane with the visual evaluation of results for the detection of tularemia IgG antibodies in hamadryas baboons at the postvaccinal period has been studied. The sensitivity of this assay has been compared with that of the passive hemagglutination (PHA) test, the microagglutination (MA) test and EIA with the spectrophotometric evaluation of results in plates. As shown in this study, EIA in the above-mentioned modification can be successfully used for the detection of tularemia antibodies in the blood serum. The sensitivity of micropoint EIA has proved to be not inferior to that of EIA in plates, while exceeding the sensitivity of the PHA test 10- to 20-fold and the sensitivity of the MA test 10- to 1,000-fold. This method is simple, reliable, highly sensitive, economic and requires no special equipment, which makes it highly promising for the diagnosis of tularemia and the evaluation of humoral immunity at the postvaccinal period.

[Dot immunoenzyme analysis in biological research]. / Tochechnyi immunofermentnyi analiz v biologicheskikh issledovaniiakh.

The modern state, general methodologic problems and the possibilities to use in biological research the dot enzyme-linked immunosorbent assay (dot-ELISA) are analysed in the review. New types of microporous polymer membranes and equipment for the application of the solid-phase reagent and performing the assay are considered. Different variants of dot-ELISA and methods for the evaluation of results obtained in the assay as well as the ways for its optimization are discussed.

[A comparative study of the immunogenicity of 2 group-B meningococcal vaccines in monkeys]. / Sravnitel'noe izuchenie immunogennosti dvukh meningokokkovykh vaktsin gruppy B na obez'ianakh.

The comparative study of two group B meningococcal vaccines manufactured in the USSR and in Cuba was made. The vaccine manufactured in the USSR contained the noncovalent compound of group B Neisseria meningitidis polysaccharide and outer membrane protein, and the Cuban vaccine contained group B N. meningitidis outer membrane proteins and group C N. meningitidis polysaccharide. The data obtained in this study indicated that both vaccines possessed immunological potency evaluated according to their capacity to stimulate the formation of bactericidal antibodies, whose level was found to increase eightfold after the immunization of monkeys in two injections. Besides, group B meningococcal vaccines did not induce the suppression of nonspecific protective activity characteristics of the body and did not stimulate the formation of autoantibodies to brain and liver tissues, which was indicative of the safety of these vaccines.

[The use of the microdot immunoenzyme method for determining antibodies to Mycobacterium tuberculosis antigens]. / Ispol'zovanie mikrotochechnogo immunofermentnogo metoda dlia opredeleniia antitel k antigenam Mycobacterium tuberculosis.

The microdot enzyme immunoassay (EIA) has been used for the determination of antibodies to M. tuberculosis protein fractions, crude antigenic preparations, PPD and old tuberculin in tuberculosis patients and healthy persons. Purified protein fractions have been found to possess the highest sensitivity and specificity in microdot EIA. The determination of antibodies to these fractions has permitted the differentiation of persons infected with M. tuberculosis from healthy ones. The use of M. tuberculosis protein fractions permits the determination of IgA and IgC in the sera of tuberculosis patients.

[Microdot immunoenzyme analysis of chorionic gonadotropin in pregnancy]. / Mikrotochechnyi immunofermentnyi analiz khorionicheskogo gonadotropina pri beremennosti.

A microdot enzyme immunoassay test system has been developed for measuring chorionic gonadotropin levels in urine samples, on the basis of affine purified antibodies to hormonal beta-subunit and antibody complex with horse radish peroxidase. The results are assessed visually with the use of a color indicator scale obtained in experiments with reference preparations containing 12.5, 50, and 200 mMU of the hormone per ml. Comparison of the intensity of microdot color permits characterizing the level of hormone secretion and diagnose pregnancy 4-5 days before the first day of expected menstruation (50 mMU/ml). The specificity, reproducibility, and sensitivity of the developed test system are not inferior to common enzyme immunoassay on the plates; the method does not involve the use of sophisticated equipment and is reagent-saving. The diagnostic accuracy of the test system is 100 percent.

[Comparative study of murine B-cells obtained by 2 different methods proliferating long-term in vitro]. / Sravnitel'noe izuchenie dlitel'no proliferiruiushchikh in vitro B-kletok myshi, poluchennykh dvumia razlichnymi metodami.

The hybridization of myeloma cells NP with lymphocytes of mice, immunized with protein isolated from Neisseria meningitidis strain Bc5 in a single injection into the spleen 3 days prior to fusion, made it possible to obtain 25-72% of hybridomas secreting antibodies to meningococcal antigens. The treatment of immune lymphocytes from these mice with the total preparations of nucleic acids, isolated by the phenol-detergent method from mouse myeloma cells NP and NS/0, induced an increase in the proliferative activity of lymphocytes; in some microcultures multilayer cell growth was observed on the bottom of the wells, whereas in the control microcultures such growth was absent. No synthesis of specific antibodies was detected in the cultures of lymphocytes whose proliferation was stimulated with nucleic acids.

[Experimental study of the immunological activity of a preparation made from the antigenic complexes of opportunistic microorganisms administered by different routes]. / Eksperimental'noe izuchenie immunologicheskoi aktivnosti preparata iz antigennykh kompleksov uslovno-patogennykh mikroorganizmov pri razlichnykh sposobakh vvedeniia.

The data obtained in the experimental study of the humoral factors of local and systemic immunity, as well as the morphofunctional changes of internal organs, after multiple subcutaneous and aerosol immunization with the combined preparation of Proteus, Staphylococcus, Klebsiella pneumoniae and Escherichia coli antigens (preparation nC-4) are presented. The subcutaneous and aerosol administration of preparation nC-4 has been found to induce an increase in the levels of antibodies to all components of the preparation in the blood serum and in respiratory tract secretions. The introduction of the preparation through the respiratory tract resulted in an earlier intensive accumulation of specific antibodies and IgA in respiratory tract secretions. The results of the quantitative cytological study of respiratory tract secretions and the morphofunctional changes of the lymphoid tissue associated with the lungs are indicative of an important role played by cell-mediated immunity factors in the formation of local postvaccinal immunity.

[Humoral factors of local and systemic immunity in the multiple peroral administration of a corpuscular pertussis vaccine]. / Gumoral'nye faktory mestnogo i sistemnogo immuniteta pri mnogokratnom peroral'nom vvedenii korpuskuliarnoi kokliushnoi vaktsiny.

Multiple oral immunization with pertussis corpuscular vaccine was shown to lead to the considerable stimulation of local and systemic humoral immunity. The data on the titers of specific and normal secretory antibodies, on the levels of IgA in washings from the oral cavity, the small intestine and the lungs, on the titers of agglutinins and hemagglutinins in the blood serum, as well as on the morpho-functional transformation of the mucous membrane and the associated lymphoid tissue in the digestive tract, are presented in their dynamics. Specific pertussis antibodies in high titers were detected in both intestinal and pulmonary washings. The multiple administration of the vaccine did not produce pathological changes in internal organs.

[Detection of antibodies neutralizing the endotoxins of gram-negative bacteria using the liposomal potentiometric method]. / Vyiavlenie antitel, neitralizuiushchikh éndotoksiny gramotritsatel'nykh bakterii, liposomal'nym potentsiometricheskim metodom.

For the first time the study of the indicator system consisting of sensitized liposomes with NaF incorporated as a marker and a fluorine-selective electrode has been made and, as a result, the possibility of the potentiometric determination of the immune lysis of liposomes in the presence of complement and specific antibodies has been demonstrated. The dissolution of the lipid components (Re-chemotype glycolipid and lipid A) in the bilayer matrix obviates the necessity for converting lipid antigens into the water-soluble state in the process of serological tests. As compared with other methods, the liposomal potentiometric method for the determination of Re-chemotype glycolipid and lipid A is highly sensitive (20-40 ng/ml), rapid, technically easy to perform, cheap and does not require large volumes of samples. The disadvantages of this analytical system are the instability of liposomes and the diffusion of fluorine ions from the internal aqueous phase of vesicules. For this reason the immunoassay can be made only within 12 hours after the preparation of sensitized liposomes incorporating the marker.

[Characteristics of the immune response to a single peroral administration of different doses of corpuscular pertussis vaccine]. / Osobennosti immunnogo otveta pri odnokratnom peroral'nom vvedenii raznykh doz korpuskuliarnoi kokliushnoi vaktsiny.

The effect of oral administrations of different doses of pertussis vaccine on the humoral and cell-mediated responses of systemic immunity and on the immunomorphological transformation of the mucous membrane of the small intestine was studied in (CBA X C57BL/6)F1 mice. On day 28 after the administration of all the tested doses of pertussis vaccine the animals were found to have a high degree of protection from the development of meningoencephalitis induced by the inoculation of Bordetella pertussis in the absence of specific hemagglutinins in their blood sera. At the same time the formation of spontaneous and immune rosette-forming cells and splenocytes was found to be inversely related to the administered dose. The immunomorphological transformation observed in the mucous membrane of the small intestine and in the lymphoid tissue associated with the small intestine was indicative of the stimulation of local immunity. The results thus obtained suggest that a single oral administration of pertussis vaccine to mice stimulates cell-mediated and humoral reactions of local immunity and induces the development of systemic cell-mediated immunity.

[Immunological activity and toxicity of a peroral vaccine made from mutant Re Salmonella minnesota]. / Immunologicheskaia aktivnost' i toksichnost' peroral'noi vaktsiny iz mutanta Re Salmonella minnesota.

The possibility of the oral use of heated corpuscular vaccine prepared from S. minnesota mutant R 595 (chemotype Re) for protection against Pseudomonas aeruginosa has been studied. Oral immunization in 3 doses, each containing 10(9) cells of the vaccine strain, has been shown to protect mice from death after the intravenous injection of P. aeruginosa culture in a dose of 5 LD50 and induce a rise in the titers of antibodies to Re-glycolipid (Re-hemagglutinins). After multiple oral administration Re-vaccine shows low acute and chronic toxicity and induces local and systemic immunological transformation.

[Persistence of pneumococci in mice]. / Persistentsiia pnevmokokka u myshei.

The sensitivity of different mouse strains to Str. pneumoniae, serotype 6, under the conditions of intraperitoneal and intranasal infection has been studied. The time course of distribution of pneumococci in the organs has been shown and differences depending on the methods of infection have been revealed. The pathomorphological and immunomorphological changes in the organs and tissues after the introduction of the infective agent by different methods have been established.

[Choline acetyltransferase in the nerve plexuses of the basilar artery of the cat brain]. / Kholinatsetiltransferaza v nervnykh spleteniiakh osnovnoi arterii mozga koshki.

Localization of choline-acetyltranspherase (CHAT 2. 3. 1. 6) and acetylcholinesterase (ACHE 3. 1. 1. 7) in the neuronal plexus of the main artery in the cat brain has been studied. It has been stated that CHAT is mainly revealed in preterminal and terminal fibres of the plexus, while AXE--along the whole length of the fibre. ACHE activity is higher than that of Chat.

[Choline acetyltransferase in the neural plexus of the main artery of cat's brain]. / Kholinatsetiltransferaza v nervnykh spleteniiakh osnovnoi arterii mozga koshki

Methods of Burt (1961, 1970) and Kása (1970) were used in order to study localization of choline acetyltransferase (ChAT 2.3.1.6) and method of Koelle to study the activity of acetylcholinesterase (AChE 3.1.1.7) in nerve plexuses of the cat's brain major artery. It has been established that ChAT is detected mainly in preterminal and terminal fibres of the plexus while AChE is detected along the total length of the fibre. The activity of AChE is higher than that of ChAT.

[Histophysiologic characteristics of the effector innervation of the arteries of the base of the brain during rat ontogenesis]. / Gistofiziologicheskaia kharakteristika éffektornoi innervatsii arterii osnovaniia golovnogo mozga v ontogeneze u krys

The development of adren- and cholinergic nervous plexuses in the brain base arteries was studied by histochemical methods of Falck and Kelle in animals and fetuses of 10-22 days, newborn rats, animals of 10, 20, 30, 40, 60 and 120 days of life and 1 and 2 years old rats. The cholinergic nerve fibres were first found in the basilar, vertebral and internal carotid arteries on the 15th and 16th days of ontogenesis. Specific fluorescence of adrenergic conductors on the same arteries is revealed somewhat later--from the 17th and 18th days of the intrauterine development. Further formation of the cholin- and adrenergic innervation of the arteries of the Willis' circle goes on synchronously. The number of nerve fibres increases with the growth of the artery diameter. The concentration of catecholamines and the activity of AChE in them gradually increases. The greatest density of nerve fibres per 1 mm2 is determined in 20-day-old rats. The number of cholinergic nerve fibres on the arteries of the brain base is equal to that of adrenergic fibres during the whole period of postnatal ontogenesis. By the 30th day the effector nervous apparatus has a definite structure. In old rats the activity of AChE and the content of catecholamines drop, the amount and concentration of nerve fibres decrease.