Vol 9, No 3 (2012)

Background. For wide introduction of the component allergy diagnostics in laboratory practice it is necessary to create methods of production, purification and verification of synthetic allergens. Major birch pollen protein Bet v 1, a representative of the superfamily PR-10, is interesting due to the high incidence of sensitization and the manifestation of cross-reactive properties. Methods. To receive a synthetic protein DNA cloning techniques were used, followed by expression in E. coli. Purification was made by metal-affinity chromatography. Evaluation of immunochemical properties was held in several tests including ELISA analysis and line-blot. Results. We have obtained high-yield bacterial strain E. coli, containing expression vector for protein Bet v 1 production. Physico-chemical and immunological properties were estimated in various test systems with the reference panel of serum. Conclusion. It was shown that the resulting synthetic analogue of protein Bet v 1 can be used to measure the presence of specific immunoglobulin E in serum samples. This allergenic component may be used for estimation of each component of the sensitization profile.

Background. To estimate the dynamics of soluble apoptosis markers in infants with atopic dermatitis for updating mechanisms of immunopathogenesis and improvement of diet therapy. Methods. We observed 66 bottle-fed infants aged 1,5—12 months old (boys -47, girls — 19) with atopic dermatitis (AD). The sensibilization to cow milk protein was revealed in all 66 infants. Detected allergen-specific IgG and IgE antibodies to cow milk protein, its fraction and goat milk protein were the reason to include infants into the 1st group and feed with hydrolyzed formula (27 infants). 39 infants in the 2nd group, who were not sensibilized to goat milk protein, were fed by goat milk based formula. Serum levels of soluble apoptosis markers (sCD153, caspase-8, sFas-L, caspase-9 and annexin-5) were measured by immunoenzyme method (ELISA). Results. The activation of signal apoptosis systems in infants with AD with increased levels of sFas-L и sCD153 was revealed. Levels of caspase-8 and caspase-9 were significantly lower than in control group, and reflected the impaired elimination of modified immmunocompetent cells. The level of annexin-5 was significantly lower in infants with AD than in control group. The estimation of the dynamics of investigated parameters during diet therapy showed significant increase of caspase-9 level in both groups. The level of caspase-8 was increased only in infants who were fed by goat milk formula. Levels of sFas-L, sCD153 and annexin-5 during diet treatment did not differ significantly between groups. Conclusion. The results showed that sCD153, caspase-8, sFas-L, caspase-9 and annexin-5 play a role in the realization of allergic inflammation in infants with AD. The diet therapy with goat milk formula promotes more physiological repair of the effectory component of the apoptosis.

Background. Improvement of control methods of treatment of mycoplasma infection in children with bronchial asthma. Methods. 52 children from 1 to 4 years old with bronchial asthma who had mycoplasma antigens in the blood were examined before treatment of mycoplasma infection, 15 patients were examined in 1,5 months after treatment. Reaction of aggregate-hemagglutination, immunofluorescence assay and the reaction of the polymerase chain reaction (PCR) were used for detection of mycoplasmas’ antigens and DNA. Results. In reaction of aggregate-hemagglutination Mycoplasma pneumoniae (Mpn) was detected in 65,4% of patients, Mycoplasma hominis (Mh) — in 32,7%, Ureaplasma urealyticum — in 50%, Mycoplasma arthritidis — in 46,2%, Mycoplasma fermentrans — in 46,2%. In more detail, we investigated Mpn and Mh. Antigens of Mpn and Mh were detected in serum significantly frequently than DNA by PCR. DNA in structure of circulating immune complexes (CIC) was detected more often than in free state. After treatment by azithromycin the number of positive tests on antigens and DNA in free state and in structure of CIC decreased. In most cases DNA found in serum of blood in free state and in structure of CIC belongs to persistent living cells of mycoplasma. Conclusion. Empowering diagnosis of mycoplasma infections in asthma in children allows to base a comprehensive approach to laboratory diagnosis, monitor treatment, predict disease course.

On time diagnostics and correct treatment of patients with PID with humoral deficiency provide a good prognosis. Despite of long experience of intravenous immunoglobulins (IVIG) usage for the treatment of these patients no strict recommendations concerning regimes of treatment exist. This article presents clinical experience about the treatment with 10% IVIG Gamunex in PID patients with immunoglobulin deficiency.