[Improving the efficacy of tuberculosis immunodiagnosis using anti-human IgG monoclonal antibodies]. / Povyshenie éffektivnosti immunodiagnostiki tuberkuleza putem primeneniia monoklonal'nykh antitel protiv IgG cheloveka.

Monoclonal antibodies (MAb) F5 to human IgG were used for creating immunoperoxidase conjugate. MAb dissociation constant was 10(-9)M-1 and the number of binding sites 1. Enzyme immunoassay (EIA) and immunoblotting showed that MAb F5 specifically recognize conformation epitope on intact human IgG molecule but not on other human immunoglobulins or denatured IgG. The resultant peroxidase conjugate with MAb F5 was used for EIA titration of antibacterial antibodies in sera from 30 patients with pulmonary tuberculosis, 28 patients with nonspecific pulmonary diseases (bronchitis and/or asthma, pneumonia), and 12 donors. For comparison similar studies were carried out with reference commercial immunoperoxidase conjugate to human IgG(H + L) manufactured at N. F. Gamaleya Institute of Epidemiology and Microbiology. Mycobacterium tuberculosis monoantigen (mol. weight 15-18 kD), affinity isolated by antibacterial MAb S4C1G4 (alpha S4C1G4), and PPD (Batch RT 45, Stattens Seruminstitut, Denmark) were used. Sensitivity and specificity of serum antibacterial antibodies were compared. The specificity of conjugate based on MAb F5 with monoantigen alpha S4C1G4 was 78.21%, sensitivity 94.50%, while those of conjugate to human IgG(H + L) were 53.30 and 76.89%, respectively (p < 0.001). For PPD the specificity and sensitivity were 56.75 and 72.33%, respectively (conjugate with MAb F5) versus 47.67 and 62.38% for conjugate against IgG(H + L), p < 0.001. Similar values were obtained in assessment of the concentrations of antibodies to alpha S4C1G4 for MAb F5 conjugate: specificity and sensitivity 47.16 and 71.56%, respectively, versus 23.67 and 95.16% for PPD (p < 0.05). No statistically significant differences between the experimental groups were detected with IgG(H + L) conjugate. We believe that specific MAb-based conjugate to human IgG will improve the efficacy of EIA as a method for the diagnosis of pulmonary tuberculosis.

[Comparative analysis of two mycobacterium tuberculosis antigens and two methodological approaches to determining serum antimycobacterial antibodies]. / Sravnitel'nyi analiz dvukh antigenov mycobacterium tuberculosis i dvukh metodicheskikh podkhodov pri opredelenii protivotuberkuleznykh antitel syvorotki v immunofermentnom analize.

The glycoprotein (15-18 kDa) antigen of Mycobacterium tuberculosis H37Rv was affinity isolated on the immunosorbent with monoclonal antibodies S4C1G4 (specific to M. tuberculosis H37Rv; Avdiyenko V. G., Kondrashov S.Yu, Lyashenko S.M.@Probl. Tuberk. 1996, v. 1, p. 6-8 (in Russian). This antigen and PPD (Batch RT 45, Stattens Seruminstitute, Denmark) that was a standard antigen were used for enzyme-linked immunosorbent assay (ELISA), by detecting serum IgG antibodies in patients with pulmonary tuberculosis, and control groups of patients with lung diseases other than tuberculosis (bronchitis and/or asthma, pneumonia) as well as healthy volunteers. The diagnostic parameters of specificity and sensitivity for titers and the same parameters for optic density (OD) (in serum dilution with maximum differences for groups of patients and donors) were compared. The new monoantigen method provided 86.11% specificity and 87.87% sensitivity, which were higher those obtained for optic density (63.89 and 80%, respectively). With PPD, the specificity and sensitivity were 58.04 and 78.78 (for the new titer method) versus 50 and 78.78% (OD data). The method error for titer determination was 10% and for standard OD determination was 27%. The new approach offers additional possibilities of enhancing the quality of ELISA for diagnosis of tuberculosis.

[Serological and immunochemical properties of carbohydrate-containing fraction from mycobacterium tuberculosis H37Rv]. / Serologicheskie i immunokhimicheskie svoistva uglevodosoderzhashchei fraktsii iz mycobacterium tuberculosis H37Rv.

Using affinity chromatography on concanavalin A (Con A) sepharose CL 6B, a carbohydrate-containing fraction was derived from Mycobacterium tuberculosis H37Rv sonicate. Surprisingly, the main component of Con A fraction was a protein having a molecular weight of a 30-kD range which is generally absent in the Con A-adsorbed fraction from the culture filtrate. ELISA by means of an antimycobacterial monoclonal antibody panel showed that the 30-kD range of Con A fraction contained antigen 85. It is suggested that the derived components antigen 85 are a glycosylated form of the proteins associated with the mycobacterial cell wall. The Con A fraction, antigen 85 (Department of Virology, Pasteur Institute, Brussels, Belgium), and PPD (Batch RT 45, Stattens Seruminstitute, Denmark) were used for ELISA determination of antimycobacterial antibody titers in the sera of 30 patients with pulmonary tuberculosis, 28 patients with nonspecific lung diseases (bronchitis and/or asthma, pneumonia), as well as in the sera of 12 healthy volunteers. The sensitivities were 46.42, 57.34, and 72.33% and the specificities were 36.97, 26.73, and 56.75% for Con A fraction, antigen 85, and PPD, respectively. The authors suggest that serodiagnostic properties of Con A fraction are extremely limited.

[Lipid peroxidation and various parameters of mineral metabolism in patients with alimentary obesity during therapy with monosodium glutamate]. / Issledovanie perekisnogo okisleniia lipidov i nekotorykh pokazatelei mineral'nogo obmena u bol'nykh alimentarnym ozhireniem pri ispol'zovanii v dietoterapii mononatriia glutamata.

Lipid peroxidation and mineral metabolism were studied in patients with III degree of obesity and bad standing of sodium restricted diet before and after treatment with including in diet of monosodium glutamate (MSG). It was established well being of MSG and absence of negative effect on loss of body mass in course of dietary treatment. MSG caused normalization of level of diene conjugates and some minerals in serum of patients.

[Systematic approach to detection of anti-idiotypic antibodies to antimycobacterial idiotype in man and animals]. / Metodicheskii podkhod k opredeleniiu antiidiotipicheskikh antitel k antimikrobakterial'nomy idiotipu cheloveka i zhivotnykh.

ELISA in the sandwich modification was developed to detect anti-idiotypic antibodies to the antituberculosis idiotype of monoclonal antibodies (MAb) S4C1G4 (anti 15-18 kDa H37Rv). Sera immunoglobulins were determined by Staphylococcus protein A capture. F[ab+]2, the MAb fragments S4C1G4 (IgG2a, kappa) and 1.3.3.B5 (IgG2a, kappa), were used as a negative control, which were not recognized mycobacterial antigens and human immunoglobulins. The anti-idiotypic antibody titers were determined in the murine hyperimmune sera during the period of immunization with the MSAb SaC1G4. The method detected anti-idiotypic antibodies in the sera of infected mice and in patients with pulmonary tuberculosis. It is suggested that this method is applicable while using diagnostic tools.

[An analysis of the specificity of anti-Mycobacterium monoclonal antibodies and of the chemical nature of the antigenic determinants with which they react]. / Analiz spetsifichnosti antimikobakterial'nykh monoklonal'nykh antitel i khimicheskoi prirody antigennykh determinant, s kotorymi oni reagiruiut.

Seven hybridomas of BALB/c mice producing monoclonal antibodies (mAb) against M. tuberculosis H37Rv have been obtained. The mAb acted against M. tuberculosis H36Rv with molecular mass 14, 17-15, 25 27 30 kD excluding mAb S5B3B8 and S3H5D7 which acted against the main antigen with 54 kD mass and 5-6 bands of antigens. Immunoglobulin isotypes of mAb were IgG2a and IgG2b. mAb can be divided into 2 groups by interaction with mycobacterial antigens in EIA: specific to M. tuberculosis complex antigens, cross-reactive to antigens of different mycobacteria. Chemical nature of antigenic determinants recognizable by a panel of mAb was investigated. Mild sodium periodate oxidation and protease digestion of mycobacterial antigens showed that mAb recognize both carbohydrate-containing epitopes (S2H6G4) and protein epitopes (S5B11E10, S5B3B8, S4C1G4) or protein and carbohydrate-containing antigenic determinants (S2F5C10, S2F2E2).

[An experimental analysis of Mycobacterium smegmatis as a possible vector for the design of a new tuberculosis vaccine]. / Eksperimental'nyi analiz Mycobacterium smegmatis v kachestve vozmozhnogo vektora dlia konstruirovaniia novoi protivotuberkuleznoi vaktsiny.

This study was inspired by WHO program for vaccine development. Our experimental model of lethal tuberculosis in mice was used to test whether rapidly growing avirulent M. smegmatis could be used as a live vaccination vector for screening rationalized M. tuberculosis cosmid library. The results suggest that M. smegmatis is not likely to possess protective activity against tuberculosis in C57B1/6 TB-susceptible mice. This negative result is satisfactory in that the immunity induced by M. smegmatis itself could not interfere with expected protection caused by the expression of M. tuberculosis genes in this vector. Our model makes it possible to find out which gene(s) of M. tuberculosis provide an ability to multiply in the host and whether virulence and persistence are controlled by the same genetic traits(s).

[Morphologic changes of peripheral lymphoid tissue and liver in experimental animals under food deprivation]. / Morfologicheskie izmeneniia perifericheskikh limfoidnykh organov i pecheni éksperimentalnykh zhivotnykh v usloviiakh golodaniia.

Morphologic examinations of liver, spleen, mesenteric lymph nodes and groups of lymphatic follicles of small intestine (Peyer's patchs) of rats in condition of starvation were shown. Starvation of animals for 48 hours and most for 96 hours was caused statistic reliable decrease of body mass, concerning mass of thymus and liver. Centrolobular fatty infiltration and decrease of RNA and glycogen contents in hepatocytes of liver were shown. Decrease of content of small lymphocytes of T-dependent zones and alteration of cell's representation in B-dependent zones were discovered.

[Effect of atherosclerotic diet containing polyunsaturated omega-3 fatty acids on level of natural antibodies to prostaglandin F2alpha in patients with ischemic heart disease, hyperlipidemia and hypertension]. / Vliianie protivoateroskleroticheskoi diety s vkliucheniem polinenasyshchennykh zhirnykh kislot omega-3 na uroven estestvennykh antitel k prostaglandinu F2alpha u bolnykh ishemicheskoi bolezníu serdtsa, giperlipoproteidemiei i gipertonicheskoi bolezníu.

The hypolipidemic, antiatherogenic and thrombolytic effects and decrease of prostaglandin F2 alpha (PG) excretion were observed in 55 patients suffering from Ischemic heart disease, hyperlipidemia and hypertension fed the antiatherosclerotic diet containing 20 g of eiconol during 55 days. The low initial level of natural antibodies to PGF2 alpha was increased significantly in blood serum. Authors consider that decrease of PGF2 alpha level under diet influence is connected not only with the change of fatty acid composition of cell membranes but also with increasing of natural antibody production.

[Characteristics of anaphylaxis in fasting rats]. / Osobennost proiavleniia anafilaksii u krys pri golodanii.

The effect of 4-day fasting was studied in male Wistar rats (200-150 g) sensitized by ovalbumin on anaphylaxis manifestations in vivo, antigen-dependent histamine secretion by mast cells in vitro, concentration of serum specific IgG antibodies. The fasting ended by a shocking allergen dose. It is shown that in the condition of food deprivation anaphylactic severity significantly lowered as well as antigen-dependent histamine secretion by mast cells. No changes were seen in the levels of specific antibodies in the serum.

[The effect of starvation on mast cell functional activity and on the content of microsomal cytochromes in the liver]. / Vliianie golodaniia na funktsional'nuiu aktivnost' tuchnykh kletok i soderzhanie mikrosomal'nykh tsitokhromov v pecheni.

Histamine secretion and ATP levels were measured in mast cells from the abdominal and pleural cavities of Wistar rat males (250-300 g) which had been starving for 2-4 days. Food deprivation had no effect on histamine levels and secretion in exposure to substance 48/80. ATP concentration got reduced from starvation day 2. Food deprivation for 2 days led to decline in the content of cytochromes P-450, P-450B, in the ratio P-450B/P-450L, but had no effect on rat liver cytochrome B5. 4-day starvation did not entail further changes.