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.

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.

[Conjugates and carbohydrate chains of alpha 1-acid glycoprotein with polyacrylamide maintain the immunomodulating activity of native glycoprotein]. / Kon''iugaty uglevodnykh tsepei alpha1-kislogo glikoproteina s poliakrilamidom sokhraniuiut immunomoduliruiushchuiu aktivnost' prirodnogo glikoproteina.

Translocation of carbohydrate glycoprotein N-chains onto soluble polyacrylamide was proposed as a method for studying the biological role of carbohydrate chains. N-linked carbohydrate chains of alpha 1-acid glycoprotein (AGP) were aminated at the reducing GlcNAc moiety and covalently attached to polyacrylamide (PAA). Thus "pseudo-AGP" was obtained where peptide core was replaced with PAA. The synthetic model mimics AGP by M(r) and carbohydrate content as well as the ratio of tetra-, tri- and diantennary and mono-, di-, tri- and tetrasialo chains. It was shown that the conjugate inhibits proliferation of lymphocytes like the parent AGP. Therefore, the property of AGP to inhibit the lymphocyte proliferation is attributed to its carbohydrate chains, whereas peptide core serves as carrier providing polyvalent interaction of multiple carbohydrate chains with cell.

[The interaction of peripheral blood leukocytes with alpha1-acid glycoprotein, its carbohydrate chains and neoglycoconjugates]. / Vzaimodeistvie leikotsitov perifericheskoi krovi s alpha1-kislym glikoproteinom, ego uglevodnymi tseliami i neoglikokon"iugatami.

The interaction of human peripheral blood leukocytes with alpha 1-acid glycoprotein (AGP), its glycoforms as well as neoglyco-conjugates representing carbohydrate chains of AGP or its fragments was studied by flow cytometry. It was shown that the main target cells for AGP as well as for conjugates of its carbohydrate chains with polyacrylamide (PAA) are monocytes and polymorphonuclear leukocytes but not lymphocytes. The interaction of AGP with monocytes and granulocytes are mediated by its carbohydrate chains: the binding of AGP with cells was inhibited by AGP, AGP oligosaccharides as well as conjugates of oligosaccharides and its fragments with PAA. The data obtained show the existence of monocyte (and granulocyte) receptors which interact with complex type sialooligosaccharides of AGP.

[Degree of proliferative response to concanavalin A of spleen cells of mice of different strains and production of interleukin-2]. / Vysota proliferativnogo otveta na konkanavalin A kletok selezenki myshei raznykh linii i produktsiia interleikina-2.

Differences in the lymphoproliferative response to Con A of spleen cells allowed one to distinguish a high responder (BALB/c and DBA/2) and low responder (C57BL/6 and CC57BR) mice. BALB/c and DBA/2 mice (H-2d haplotype) produced interleukin 2 better, than C57BL/6 and CC57BR mice (H-2b haplotype). However acceptance of interleukin 2 was better in BALB/c and C57BL/6, than in DBA/2 and CC57BR mice. Summarizing these facts the authors suppose that the differences in interleukin 2 production and acceptance play an important role in the height of lymphoproliferative response.

Pharmacokinetics of alkylating metabolites of cyclophosphamide in different strains of mice.

Determination of the amount of alkylating (NBP) metabolites of cyclophosphamide (CP) in blood serum of BALB/c and DBA/2 mice has revealed that the shape of the curve describing the accumulation of NBP-metabolites, depends on the administered drug dose and animals' genotype. The experimental data obtained made it possible to suggest an improved pharmacokinetic model which takes into account a possible switching in of factors conditioning non-linear changes in the intensity of the accumulation processes in blood and elimination of CP alkylating metabolites from blood. The effect of different CP alkylating metabolite kinetics in BALB/c and DBA/2 mice on a different sensitivity of these mice to the immunodepressive action of CP in vivo, has been discussed.

[Genetic control of interleukin-2 production in inbred mice]. / Izuchenie geneticheskogo kontrolia produktsii interleikina-2 u myshei inbrednykh linii.

BALB/c mice (H-2d haplotype) produced IL 2 better than C57BL/6 mice (H-2d haplotype). However genetic analysis in F2 generation demonstrated independent segregation of the IL 2 production intensity and H-2 haplotype. Investigation of the IL 2 production intensity in BALB/c, C57BL/6 and their F1 and F2 generation revealed that this was controlled by only one gene.

Immunosuppressive action of cyclophosphamide in mice: contribution of some factors to determination of strain differences.

Mechanisms of strain differences in susceptibility to cyclophosphamide (CP) has been investigated with BALB/c, DBA/2, CC57BR, and C57BL/6 mice. It was shown that the final immunosuppressive effects of CP is a result of a combination of such factors as the level of CP alkylating metabolites in the bloodstream and the sensitivity of the target cells to antiproliferative action of these metabolites. The importance of the immune status peculiarities has been discussed.

[Comparative study of mechanisms of antiproliferative action of low concentrations of mafosfamide and cyclosporin A]. / Sravnitel'noe izuchenie mekhanizmov antiproliferativnogo deistviia nizkikh kontsentratsii mafosfamida i tsiklosporina a.

Exact mechanisms of Mf and CsA effect have been investigated in BALB/c, DBA/2, CC57BR, and C57BL/6 mice. Different sensitivity of spleen cells of mice of different strains to Mf and CsA antiproliferative effect has been demonstrated. These interstrain variations have not been related with an inhibition of interleukin-2 (IL-2) release but, perhaps depended on the IL-2 receptor (IL-2R) expression differences. The p75 chain of IL-2R as a possible target for Mf, has been discussed.

Spectrum of anti-measles immunoglobulin G subclasses in convalescents after measles.

A simple semiquantitative method for measuring anti-measles IgG subclasses is developed on the basis of commercial diagnostic test system for measurements of anti-measles IgG and a kit of peroxidase-labeled monoclonal antibodies to human IgG subclasses. During the acute phase of the disease specific antibodies are presented mainly by IgG2 antibodies, while in subjects with a history of measles more than 10 years before 2 subgroups were detected, which responded by production of IgG2 or IgG1 subclasses.

[Antiprolifirative effect of chlorbutin and sensitivity of patients with glomerulonephritis to this drug]. / Antiproliferativnoe deistvie khlorbutina i chuvstvitel'nost' bol'nykh glomerulonefritom k terapii etim preparatom.

Children suffering from different forms of glomerulonephritis were examined for the sensitivity of peripheral blood lymphocytes to the antiproliferative action of chlorambucil in vitro. All the patients were given chlorambucil treatment in accordance with a 4-component scheme. As a result, in 15 patients (group I), a complete clinicolaboratory remission was attained. In 8 patients, (group II), the treatment results were estimated as a partial remission and in 8 patients (group III), the treatment turned out ineffective. In 14 patients of group I, the sensitivity of peripheral blood lymphocytes to the antiproliferative action of chlorambucil was viewed as high whereas in 7 patients of group III as low. Peripheral blood lymphocytes from group II patients had a low sensitivity to chlorambucil. The prognostic value of the laboratory test suggested is under discussion.