Cytokine profile in children with inflammatory bowel disease.

Crohn's disease (CD) and ulcerative colitis (UC) are inflammatory bowel diseases (IBD) characterized by immune reactivity against microbial and auto-antigens. This work was designed to study the cytokine profile in blood serum and coproextracts of children with CD and UC. The studied patients consisted of 17 children with CD (group I), 17 children with UC (group II), and 18 controls with intestinal dysbiosis (group III). The diagnosis of UC and CD was based on accepted clinical and endoscopic criteria. The levels of 13 cytokines (IL-1ß, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-17A, TNF-α, TGF-ß, and IFN-γ) were determined in blood sera and coproextracts of the patients and controls using the BioPlex technology. The level of IL-17A was significantly increased and that of TGF-ß was significantly decreased in the blood serum of the patients with IBDs. Changes in the cytokine profile in the coproextracts affected the wider spectrum of cytokines. The levels of proinflammatory cytokines (IL-2, IL-4, IL-6, IL-12p70, TNF-α, and IFN-γ) were increased 6-9-fold, whereas the level of the anti-inflammatory cytokine IL-10 was increased 3-fold. The cytokine balance was shifted to the proinflammatory cytokines. The TGF-ß level was increased 9-fold and that of IL-17A was increased 3-fold. Thus, the cytokine profile in the coproextracts was more informative than that of the blood serum. The determination of cytokines in coproextracts is simple and noninvasive.

[Evaluation of the immunological activity and safety of group B meningococcal vaccine prepared from a natural complex of specific polysaccharide and outer membrane proteins]. / Otsenka immunologicheskoi aktivnosti i bezopasnosti meningokokkovoi gruppy B vaktsiny iz prirodnogo kompleksa spetsificheskogo polisakharida i belkov naruzhnoi membrany.

Immunological activity and safety of group B meningococcal vaccine prepared from a natural complex of specific polysaccharide and outer membrane proteins were under study. The immunological safety of the vaccine was evaluated by the absence of antibodies to denaturated and native DNA (d-DNA and n-DNA). As shown with the use of the enzyme immunoassay (EIA), the administration of the vaccine did not induce antibody formation to d-DNA and n-DNA during the observation period. The titer of bactericidal antibodies in the immune bacteriolysis assay (IBA) to the vaccine strain B:2b:P1.2 after immunization increased four-fold and greater in 80% of the vaccinated persons. The significant increase of bactericidal antibodies to heterologous strains B:2a:P1.2 and B:15:P1.7 was registered in 20-30% of the vaccinees, respectively. A month after the repeated vaccination an increase in specific IgG antibodies to the complex antigen was found to occur according to EIA results. The use of RIB made it possible to evaluate the preventive activity of group B meningococcal vaccine as a whole and to suppose that the vaccine induced mainly type-specific response.

[The molecular parameters of a meningococcal serogroup B protein-polysaccharide vaccine]. / Izuchenie molekuliarnykh parametrov meningokokkovoi belkovo-polisakharidnoi vaktsiny serogruppy B.

Methods for the evaluation of the molecular parameters of B polysaccharide (B-PS) in meningococcal protein-polysaccharide vaccine of group B are proposed. The comparison of two proposed methods, the passive hemagglutination inhibition test and rocket immunoelectrophoresis (RIEP), has shown that the latter method has the highest degree of correlation with the chemical method of the detection of B-PS, which is often hindered by lactose added as a bulking agent. RIEP may be recommended for the standardization and control of the commercial preparations of group B meningococcal vaccine. B-PS contained in the vaccine is known to be in a highly polymeric state: its yield is 66.4 +/- 1.7% to Kd = 0.25 and 85.3 +/- 1.2% to Kd = 0.5. B-PS contained in the prepared vaccine has been found to be highly stable. Outer membrane proteins of meningococci, forming a noncovalent complex with capsular B-PS, seem to stabilize its structure and prevent the depolymerization of molecules.

[The effect of the conditions for Neisseria meningitidis cultivation on the protein composition in the protein-polysaccharide complex--the base of meningococcal serogroup B vaccine]. / Vliianie uslovii kul'tivirovaniia Neisseria meningitidis na sostav belok v belkovo-polisakharidnom komplekse--osnove meningokokkovoi vaktsiny serogruppy B.

New data concerning the influence of cultivation conditions on the synthesis aimed at producing immunologically active antigens of group B Neisseria meningitides were obtained. The combination of conditions (deficiency in ions of iron, pH of the medium, growth phase of the culture) capable of essentially increasing (9- to 28-fold) the content of iron-dependent protein with a mol. wt. of 75-76 kD, was selected. At the same time the content of other immunologically active proteins with mol. wt. of 92-94 kD, 100-102 kD and 141-144 kD perceptibly increased (1.5 to 2.2-fold). Group B meningococcal vaccine prepared on the basis of native protein-polysaccharide complex with a higher content of the above-mentioned species-specific proteins may be expected to have a wider spectrum of immunological action.

[The relationship between the immunological efficacy of a dried meningococcal group-A polysaccharide vaccine and the molecular parameters of the group-A polysaccharide]. / Vzaimosviaz' immunologicheskoi éffektivnosti sukhoi polisakharidnoi meningokokkovoi gruppy A vaktsiny i molekuliarnykh parametrov gruppovogo A-polisakharida.

This work deals with the problem of relationship between the molecular parameters of group A meningococcal polysaccharide and its immunological effectiveness for laboratory animals and humans. The depolymerization of group A polysaccharide contained in the vaccine leads to a decrease in its capacity of inducing the production of hemagglutinating (19S and 7S) and bactericidal IgA antibodies in humans, as well as inducing an increase in the number of cells producing IgA antibodies in the spleen of immunized mice and the appearance of circulating IgA antibodies in their sera. As shown in this investigation, fully developed immune response to group A meningococcal vaccine may be achieved in humans only if the content of group A high-molecular polysaccharide in the vaccine is not less than 70%. Mice have been recommended as an experimental model for the prognostication of the effectiveness of meningococcal polysaccharide vaccines and for their control in the process of manufacture instead of currently used titration of bacteriolysins in the sera of immunized humans.

[The protective activity of polyclonal and monoclonal antibodies to the lipopolysaccharide of Neisseria meningitidis serogroup A in in vivo experiments]. / Protektivnaia aktivnost' poliklonal'nykh i monoklonal'nykh antitel k lipopolisakharidu Neisseria meningitidis serogruppy A v opytakh in vivo.

The protective activity of the sera of mice immunized with the preparations of native and detoxified N. meningitidis lipopolysaccharide (LPS), group A, as well as with monoclonal antibodies to N. meningitidis antigens, groups A and B, was studied on the mucin model of meningococcal infection. The study showed that the maximum level of anti-LPS antibodies in mice was observed on day 7 after the injection of LPS. Immune sera obtained from mice were capable of protecting the animals from fetal meningococcemia induced by N. meningitidis strains of homologous and heterologous groups. As shown by the results of this study, the alkaline treatment of N. meningitidis native LPS did not decrease the protective properties of antibodies. The monoclonal antibodies under study were found to possess high preventive activity in mice challenged with N. meningitidis, groups A and B. Anti-LPS monoclonal antibodies showed greater protective activity than antipolysaccharide monoclonal antibodies.

[The protective activity of the detoxified lipopolysaccharide of Neisseria meningitidis serogroup A in in vivo experiments]. / Protektivnaia aktivnost' detoksitsirovannogo lipopolisakharida Neisseria meningitidis serogruppy A v opytakh in vivo.

The immunogenic potency, toxicity, homologous and heterologous protective activity of lipopolysaccharide preparations obtained from serogroup A N. meningitidis (LPS A) were studied in animal experiments. These preparations were shown to possess very high protective activity. The alkaline treatment of native LPS A decreased the toxicity of the preparation almost 20 times and did not affect its immunogenic potency. Detoxified LPS A was capable of protecting mice from fatal meningococcemia resulting from infection with N. meningitidis strains, serogroups A, B and C; the adsorption of the preparation on aluminium hydroxide did not affect its protective activity. In view of the properties of detoxified LPS A revealed in this investigation, it may be regarded as a possible vaccinal preparation.

[Evaluation of the effectiveness of group-A and -C meningococcal polysaccharide vaccines by molecular parameters]. / Otsenka éffektivnosti meningokokkovykh polisakharidnykh vaktsin grupp A i C po molekuliarnym parametram.

Chromatographic properties of meningococcal polysaccharides and vaccines of serogroups A and C prepared at the G. N. Gabrichevskii Moscow Research Institute of Epidemiology and Microbiology were studied during determination of their molecular parameters by gel filtration on sepharose 4B. It was shown that the character of the polysaccharide elution was not affected by the composition of the eluent and pH changes within the ranges of 6.5-8.5. The ionic strength of the solvent should be at least 0.2 mol/l. Immunochemical methods such as countercurrent and rocket immunoelectrophoresis were standardized and recommended for quantitative evaluation of the content of meningococcal polysaccharides of groups A and C in the eluates. Rocket immunoelectrophoresis was shown to be more accurate and reproducible.