The gene therapy of collagen-induced arthritis in rats by intramuscular administration of the plasmid encoding TNF-binding domain of variola virus CrmB protein.

Wistar rats with collagen-induced arthritis were intramuscularly injected with the recombinant plasmid pcDNA/sTNF-BD encoding the sequence of the TNF-binding protein domain of variola virus CrmB protein (VARV sTNF-BD) or the pcDNA3.1 vector. Quantitative analysis showed that the histopathological changes in the hind-limb joints of rats were most severe in the animals injected with pcDNA3.1 and much less severe in the group of rats injected with pcDNA/sTNF-BD, which indicates that gene therapy of rheumatoid arthritis is promising in the case of local administration of plasmids governing the synthesis of VARV immunomodulatory proteins.

Relationship between interleukin-1 type 1 and 2 receptor gene polymorphisms and the expression level of membrane-bound receptors.

The biological activity of the multifunctional cytokine interleukin-1 (IL-1) is mediated by its receptors. The aim of this study was to determine if an association exists between single nucleotide polymorphisms (SNPs) in the IL-1 type 1 and 2 receptor genes (IL1R1 and IL1R2) and the expression level of membrane-bound IL1Rs on subpopulations of mononuclear cells or serum levels of soluble IL-1 receptors. It was observed that healthy individuals with the genotype TT in SNP rs2234650:C>T had a lower percentage of intact CD14(+) monocytes expressing IL1R1 on their surface. The SNP rs4141134:T>C in IL1R2 has also been associated with the percentage of intact CD3(+) T cells expressing IL1R2. Furthermore, individuals carrying the CC allele of SNP rs4141134:T>C and the TT allele of SNP rs2071008:T>G in IL1R2 had a lower density of IL1R2s on the surface of CD14(+) monocytes in lipopolysaccharide (LPS)-stimulated PBMC cultures. In summary, this study demonstrated that IL-1 receptor gene polymorphisms could be one of the factors influencing the expression of membrane-bound IL-1 receptors (IL1R) on immunocompetent cells.

Tissue-specific expression of splice variants of human IL-4 and IL-6 gene mRNA.

The spectrum of alternatively spliced IL-4 and IL-6 gene mRNA was studied in peripheral blood mononuclears from healthy donors and in human fetal tissues. It was found that the expression of alternatively spliced IL-4 and IL-6 gene mRNA in fetal tissues is tissue specific and that hemopoiesis- and immunopoiesis-related tissues differ by the amount of IL-4 and IL-4δ2 mRNA. An mRNA variant IL-4alt3 carrying partial exon 3 deletion was for the first time identified in human mononuclear cells.

Production of TNF-α and IL-1ß by peripheral blood mononuclear cells in carriers of different allele variants of the gene.

Associations of cytokine production by mononuclear cells and the TNF-α genetic polymorphism in positions -238, -308, -376, -857, -1031, and of IL-1ß in positions -31 and +3954 were studied. The data on distribution of allele and genotype incidence and on the level of spontaneous and mitogen-induced production of these cytokines by donor mononuclear cells demonstrated a statistically significant association of TNF-α production by mononuclear cells with polymorphic variants of the gene promoter regions -238>A (rs361525) and -857C>T (rs1799724). Carriers of -238GG/-308GG/-857CC/-1031NC genotype were characterized by low production of TNF-α, while carriers of -31TT/+3954CT genotype were characterized by low IL-1ß stimulation index in response to mitogen in comparison with carriers of other genotype combinations.

Evaluation of the expression of isoforms of stem cell factor mRNA in fetal tissues and mononuclear cells at different stages of human development.

We studied the expression of isoforms of stem cells factor mRNA forming as a result of alternative splicing. Both isoforms of stem cell factor mRNA forming as a result of alternative splicing were found in different fetal tissues. Changes in the expression of alternative isoforms of stem cell factor in peripheral blood mononuclear cells were demonstrated from the prenatal and neonatal periods to adult organism.

Ontogenetic features of the expression of mRNA isoforms for leukemia-inhibitory factor in human fetal tissues and mononuclear cells.

The expression of leukemia-inhibitory factor mRNA in human fetal tissues and mononuclear cells was studied during ontogeny. The expression of mRNA isoforms for leukemia-inhibitory factor was tissue-specific at the stage of prenatal development. The transition from antenatal and neonatal development to the postnatal period was accompanied by a decrease in the expression of mRNA isoforms for leukemia-inhibitory factor in mononuclear cells.

Effects of recombinant IL-4delta2 on human peripheral blood mononuclears.

In human cells, expression of IL-4 gene involves alternative mRNA splicing. IL-4delta2 splice variant is an antagonist of full-length IL-4 protein and blocks its effect on the functional activity of immunocompetent cells. The effect of recombinant IL-4delta2 on cytokine-producing activity of human peripheral blood mononuclear cells is shown for the first time.

Diurnal dynamics of cytokine spectrum produced by immunocompetent cells of intact mice.

The production of cytokines by immunocompetent cells of intact mice was studied in various phases of the diurnal cycle. The ratio of produced cytokines was shown to differ in various phases of the diurnal cycle. The correlation between spontaneous production of various cytokines disappeared at 20:00, which reflects the development of physiological immunosuppression due to evening increase in physical activity of mice and corresponding changes in neuroendocrine status. Our results illustrate the existence of diurnal variations in intercellular interactions in the immune system. These data should be taken into account in the study of the mechanisms for cytokine immunoregulation and development of new schemes for cytokine immunocorrection.

Proliferative activity of cells in mouse thymus and spleen under different diurnal regimens of interleukin-2 administration.

Circadian variations in spontaneous and concanavalin A-stimulated proliferation of mouse thymocytes and splenocytes were studied after administration of recombinant interleukin-2 at different times of day. Differences were revealed in the effect of morning and evening treatment with the cytokine. The time of injection corresponded to various phases of the natural circadian rhythm of endogenous interleukin-2 production, which probably contributes to diurnal differences in the influence of this cytokine.

Production of cytokines by immature erythroid cells derived from human embryonic liver.

It is well known that regulatory interactions between hematopoietic and lymphoid cells are mediated by different mediators. The cells of erythroid lineage are not an exception and have a regulatory effect on hemato- and immunopoiesis that can be mediated through the production of cytokines i.e. by soluble factors - a universal mechanism for cell regulation in hematopoietic and immune systems. It has been previously shown that erythroid progenitor cells from mice express mRNA of cytokines such as IL-1 alpha and beta, IL-4, IL-6, IFN-gamma, GM-CSF and TGF-beta. In this report we present the results of the production of the main immunoregulatory cytokines by erythroid cells derived from human embryonic liver. It was revealed that the cell population enriched with erythroid progenitors, isolated from human fetal liver, can produce IL-1 beta, IL-2, IL-4, IL-6. The levels of production of cytokines by immature erythroid progenitor cells is compared to the levels of corresponding cytokines produced by mitogen-stimulated peripheral blood mononuclear cells. The production of these cytokines changed quantitatively under the effect of erythropoietin, and are correlated with the expression of differentiation markers of erythroid cells such as AG-EB and Glycophorin A. The role of cytokine production by erythroid cells in hemato- and immunopoiesis and the mechanisms of self-regulation of proliferation and differentiation of erythroid progenitor cells is discussed.

[Level of Tx1- and Tx2-type cytokines in blood sera of hepatitis C patients]. / Soderzhanie tsitokinov Tx1 i Tx2 tipa v syvorotke krovi bol'nykh gepatitom C.

The content of cytokines of type Tx1 (IL-2 and IFN-gamma) and type Tx2 (IL-4) in blood sera of 132 patients with hepatitis C and the combined form of hepatitis B + C was studied. For control, blood sera taken from healthy donors were used. A significant increase, in comparison with the control, in the content of IL-4 in all subgroups of the patients was registered. The content of IFN-gamma reached the maximum level in patients with acute hepatitis C with the positive result of the polymerase chain reaction (PCR) for the virus (216.4 and 46.4 pg/ml respectively) and was somewhat lower in acute hepatitis C with the negative PCR-result (77.7 and 9.6 pg/ml), mean while in the chronic course of hepatitis C these data were within the limits of control values irrespective of the results of PCR. In case of mixed infection in the acute clinical form a significant increase in the concentration of IFN-gamma (34.4 pg/ml) in comparison with the control (25.3 pg/ml) was observed. The content of IFN-gamma in patients with acute hepatitis C and the positive result of the test for NS antibodies also reached the maximum level (207.3 and 42.7 pg/mg respectively). But in contrast to hepatitis C in the acute form with the negative results of PCR in patients with hepatitis C in the acute form and the negative results of the NS test these data were within the limits of control values, as well as in the chronic course of hepatitis C irrespective of the results of the NS antibodies serum test. In case of mixed infection a significant increase in the concentration of IFN-gamma was registered in the subgroup of patients with the acute form of NS+ (39.9 pg/ml). The data obtained in this study were indicative of significant changes in the serum profile of serum cytokines of types Tx1 and Tx2 in different forms and courses of virus hepatitis. This makes it possible to believe that the chronization of the process was associated with the prevalence of the Tx2 function.