Detection of Small Subsets of CD4+ Lymphocytes with SmartFlare Nanoprobes.

SmartFlare technology allows detection of mRNA in single living cells. We studied the possibility of using SmartFlare nanoprobes for detection of small subsets of CD4+ lymphocytes. It was found that SmartFlare allows detection of transcriptional master regulators of major CD4+T helper subsets in living human lymphocytes. Nanoprobes labeled with Cy5 fluorophore were better detected by flow cytometry than nanoprobes labeled with Cy3. Appropriate time of lymphocyte incubation with SmartFlare probes was 24 h.

[STRUCTURAL ABNORMALITIES OF THE OBSTRUCTED URETEROVESICAL JUNCTION].

The article presents a morphological study of UVJ removed during surgery in 110 children with stage III hydronephrosis. Morphological changes in resected UVJs indicate a predominance of sclerotic transformation accompanied by different types of dysplasia.

[Features of patients with the multiple sensitization diagnostics before allergen specific immunotherapy will be set and the assessment of the therapy results with the laboratory methods].

BACKGROUND: Due to the wide expansion of atopy, its early beginning, variety of forms, difficulty of specific pathogenetic treatment, and also high cost of in vitro researches there is a need of diagnostic test systems development and optimization. AIM: To make the assessment of atopy diagnostics efficiency in children with a multiple allergy and the analysis of specific immunotherapy (ASIT) influence on immune markers level in serum for a choice of the most significant predictive indicator. PATIENTS AND METHODS: 457 children (mean age 8,9 ± 4,3 years) with pollinosis symptoms that prevalence in spring period were tested with birch pollen allergens extract by detecting allergen-specific immunoglobulin E (sIgE) levels. Than patient witch showed positive sIgE level (243 children) were tested with expanded set of plant allergens (birch, alder, hazel, oak pollen, allergens of Rosaceae family and carrot) and set of birch pollen allergocomponents (Bet v1, Bet v2, Bet v4, Bet v6). From them 32 patients were treated with allergen-specific immunotherapy. Immunological assays were performed by indirect immunofluorescent method on ImmunoCAP250 (Sweden). RESULTS: It was shown that birch allergens sIgE antibodies detection in patients with pollinosis allows to estimate sensitization degree to allergens of related trees and could predict their quantitative values. The oak allergens sIgE level is a good predictive marker of sIgE level to food plant derived allergens. And apple allergens sIgE concentration is closely assotiated with sIgE to fruit allergens of Rosacea family. Detection of sensitization to minor allergens in patient influences on therapy efficacy prognosis. CONCLUSION: sIgE detection to limited number of allergens (birch-oak-apple) is effective to sIgE value assessment in patient with allergy to plant causing allergens cross reactivity. Component-divided in vitro diagnostics directed on reveal of sensitization caused by minor allergens, is actual at the answer a question about ASIT validity and its efficiency. Component-divided in vitro diagnostics directed on reveal of sensitization caused by minor allergens, is actual at the answer a question about ASIT validity and its efficiency. Significant results of the therapy are shown after double course ASIT that also allows to reduce considerably production of sIgE antibodies to significant allergens, and cross reacting plant food allergens.

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.

[Immunophenotypic characteristics of blood lymphocytes and expression of cytokine receptors in newborn infants].

Formation of the immune system during the neonatal period was studied in 88 infants aged 3 days-1 month (gestation age 28-41 weeks) with the primary diagnosis of cerebral ischemia. The patients were allocated to 4 groups based on chronological and gestational age at the moment of examination and a reference group. A total of 16 umbilical blood samples were available for analysis. Phenotype of blood lymphocyte was identified by flow cytofluorimetry with differentiation between subpopulations of T and B lymphocytes and detection of membrane receptors of IL-2, 8, and 10. T-cell subpupulation was found to be enriched in CD3+CD4+CD8+ lymphocytes. Content of B1 lymphocytes was equally high in full-term and premature infants. The latter showed enhanced expression of IL-2 receptor, CD122, by the end of the first month of life. Expression of receptors of IL-8 (chemotaxis factor) and IL-10 was high in lymphocytes of children of all groups since the onset of the neonatal period. It is concluded that comprehensive study of membrane cytokine receptors is of interest for better understanding mechanisms underlying maturation of the immune system in children of different age groups.

[In vitro cell adhesion and integrin expression by calcium ionophore-treated mononuclear cells from patients with acute myeloid leukemia]. / Adgezivnye svoistva i ékspressiia integrinov kletkami bol'nykh ostrymi mieloidnymi leikozami, stimulivannymi v kul'ture ionoforom dlia ionov kal'tsiia.

As shown elsewhere, cultured acute myeloid leukaemia blasts acquire certain characteristics of dendritic cells upon stimulation with cytokines and calcium ionophore. The ability of leukaemia-derived dendritic-like cells to express immune costimulatory molecules and dendritic cell marker CD83 has been extensively investigated. Although migratory capacity is a major attribute of dendritic cells, the ability of in vitro modified blasts for adhesion, chemotaxis and homing remain elusive. In the present paper, we show that after stimulation with calcium ionophore acute myeloid leukaemia blasts as well as normal dendritic cell precursors demonstrate increased capacity of binding fibronectin and denatured collagen. The expression pattern of integrins on dendritic-like leukaemic cells in general closely resembles that of monocyte-derived dendritic cells, however, variation in cell properties isolated from blood of individual patients are observed.

[Membrane markers and interleukin-2 receptor of splenic lymphocytes in children with aplastic anemia: detection of active subpopulations]. / Membrannye markery i retseptor k interleikinu-2 limfotsitov selezenki u detei s aplasticheskoi anemiei: vyiavlenie aktivnykh sub- populiatsii.

A total of 16 children with aplastic anemia were investigated, among them 6 girls and 10 boys, the mean age 10.5 years. Splenectomy was included into the complex of treatment along with glucocorticoid and transfusion-replacement therapy of all the children. The membrane markers of splenic lymphocytes were assayed in the indirect immunofluorescence test with monoclonal antibodies of ICO and OKT series. The mean parameters did not significantly differ from those of subpopulations of splenic lymphocytes in the control group described in the literature. An individual analysis made in 8 patients (50%) has revealed active lymphocyte populations in the spleen: in the group of patients with a favourable prognosis to the disease an increase in the percent of B-cells was recorded in 2, OKT8(+)-lymphocytes in 1 patient; a rise in the percent of cells with receptors to interleukin-2 in 3 patients (from 25 to 32%); in the group of patients with severe aplasia an increase in the percent of activated HLA-DR(+)-lymphocytes was recorded in 2, and OKT8(+)-lymphocytes in 1 patient. All the patients responded well to the treatment. It has been suggested that in all cases the splenectomy effect is associated with the removal of immunoactive lymphocytes and can serve as an equivalent of the immunosuppressive therapy. The effect of splenectomy in the rest cases has been obscure.