Cytokine-induced differentiation and proliferation of human T lymphocytesin vitro: Effects of interleukin 2 and interleukin 6

The contents of CD8⁺, CD4⁺CD8⁺, CD3⁺HLA-DR⁺, CD⁺INF-γ⁺ T cells, and natural killers (CD16⁺56⁺) and NK/T cells (CD16⁺56⁺CD3⁺) increase after 7-day culturing in the presence of interleukin-2. The number of apoptotic cells and cells in S-, and G₂+M phases of the cell cycle also increased. Interleukin-6 predominantly induced proliferation of CD3⁺HLA-DR⁺ T cells and G₂+M mitotic cells. Translated fromByulleten' Eksperimental'noi Biologii I Meditsiny, Vol. 129, No. 6, pp. 667–671, June, 2000     

Experession of human interleukin-10 inEscherichia coli cells

The human interleukin-10 gene has been synthesized by the chemical enzymatic method. Vectors for cytoplasmic and periplasmic expression of recombinant interleukin-10 have been obtained inE. coli cells. A high level of protein expression was found to be characteristic of only recombinant strains producing interleukin-10 as “fused” protein (fused with the N-terminal fragment of interleukin-3). Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 3, pp. 324–327, March, 1995 Presented by Yu. A. Romanov, Member of the Russian Academy of Medical Sciences). (Address for correspondence: Nauchnyi Pr. 8, 117246 Moscow, fax 331-01-01.     

RecombinantEscherichia coli strains provide high-level expression of human interleukin-3 and interleukin-4

A number of expression vectors carrying the genes coding for interleukin-3 and interleukin-4 are constructed. The influence of the promoter type, the ribosome-binding site, and the number of plasmid copies on the expression of recombinant proteins and the stability of producing strains is studied.Escherichia coli strains providing a stable high level production of human interleukin-3 and interleukin-4 are obtained. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 1, pp. 83–85, January, 1995 Presented by Yu. A. Romanov, Member of the Russian Academy of Medical Sciences     

Time course of the cytotoxicity of blood mononuclears in patients with bladder cancer during endolymphatic immunotherapy with lymphokine-activated killers and recombinant interleukin-2

A specific stimulation of mononuclear cellsin vivo was observed after 3 or 4 endolymphatic injections of autologous lymphokine-activated killers and recombinant interleukin-2 in male patients with disseminated bladder cancer. This activation presented as an increase of the cytotoxicity of mononuclears towards target cells of bladder carcinoma. A statistically reliable increase of natural killer activity was observed, and in one patient a reliable increase of the cytolytic activity of mononuclear cells against Mel-1 target cells. After 1 or 2 injections the activity of natural killers increased to 80–90% for an initial level of 23–50%. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 121, N ^o 2, pp. 188–191, February, 1996 Presented by N. N. Trapeznikov, Member of the Russian Academy of Medical Sciences     

Comparative study of the effects of recombinant tumor necrosis factor and synthetic peptides corresponding to its fragments on phagocytosis mediated by Fc- and MF-receptors

The effects of human recombinant tumor necrosis factor and its peptides on the macrophage-like cells of continuous cell line P388D1 were studied. One of the peptides, 123–131, was found capable of not only mimicking, but also of blocking the effects of recombinant tumor necrosis factor. The results permit us to tentatively identify the 123–131 site of the factor molecule as being responsible for its activating effect on macrophages. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 121, N ^o 3, pp. 304–308, March, 1996 Presented by I. P. Ashmarin, Member of the Russian Academy of Medical Sciences     

Inhibition of IL-2-Dependent proliferation of human lymphocytes and of the expression of interleukin-2 receptors by synthetic peptide fragments of α-2 interferon

It is shown that biologically active peptides inhibit the concanavalin A-induced generation of cells which may proliferate in response to exogenous recombinant interleukin-2. Determination of the number of CD25-containing cells showed that synthetic peptides are able to lower not only the number of CD25-containing cells but the level of its expression as well. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 117, N ^o 1, pp. 65–68, January, 1994 Presented by I. P. Ashmarin, Member of the Russian Academy of Medical Sciences     

Ultraviolet photomodification of the functional state of leukocytes under conditions of experimental endotoxemia

Ultraviolet irradiation of peripheral blood in therapeutic doses corrects the immuno-suppressive effect of staphylococcal toxin. The main immunoregulatory effect of ultraviolet irradiation is associated with the production of endogenous immune transmitters by blood cells such as interleukin-1 and tumor necrosis factor. Ultraviolet irradiation increases the content of CD3⁺ and CD4⁺ lymphocytes and restores the natural killer activity. The last parameter is proposed as a convenient criterion of the efficiency of quantum hemotherapy. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 123, No. 6, pp. 687–689, June, 1997     

Synthesis and properties of the immunotoxin CD5-ricin

Synthesis and properties of an immunotoxin produced by conjugating ricin with a novel monoclonal antibody (IgG3 of the ICO-104 class) are described. Cytolytic activity of the synthesized immunotoxin, determined by two independent methods and expressed in LD₅₀, is 0.3–0.6×10⁻⁷ M. Its specificity for target cells containing the CD5 antigen is shown. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 1, pp. 76–79, January, 1995 (Presented by Yu. A. Romanov, Member of the Russian Academy of Medical Sciences)     

Expansion of CD14+CD16+ peripheral monocytes among patients with aseptic loosening

Objective and design  In this study, we have investigated the relevance of peripheral blood inflammatory CD14⁺CD16⁺ monocytes phenotype to patients with aseptic loosening (AL). Material and treatment  Immunophenotypes of monocytes were examined among patients with AL (n = 43), patients with mechanical loosening (ML, n = 30), patients with stable implant (SI, n = 16), and patients with osteoarthritis (OA, n = 17) using flow cytometry. Methods  Immunological assay was used to measure TNF-α and IL-1β levels in both sera and culture media of implant wear stimulated CD14⁺CD16⁺ and CD14⁺⁺CD16⁻ monocytes. Periprosthetic tissues were collected during surgery for histological assessment. Results  The frequency of CD14⁺CD16⁺ monocytes showed significant increase in AL patients than in ML, SI, and OA patients. A positive association was found between the subpopulation of CD14⁺CD16⁺ monocytes and plasma TNF-α and IL-1β level in AL patients. Furthermore, a positive correlation existed between the subpopulation of CD14⁺CD16⁺ monocytes and the total histopathology score. Conclusion  The results indicate that CD14⁺CD16⁺ monocytes represent a sensitive marker for the disease activity of AL, and may serve as an effective prognostic index to identify total joint replacement recipients who are at increased risk for osteolysis and progression of AL.     

Involvement of fibronectin and interacting serum components in the regulation of activity of human natural killer cells

The cytotoxic activity of natural killer cells and the intensity of conjugate formation are studiedin vitro in the natural cytotoxicity reaction against³H-uridine-labeled human erythromyeloleukotic cells K-562 in the presence of fibronectin, γ-globulin, and fibronectin/γ-globulin combination. It is demonstrated that fibronectin does not change natural cytotoxicity, γ-globulin increases the activity of human natural killer cells, and the fibronectin — γ-globulin combination increases both the intensity of conjugate formation and the cytotoxic activity of natural killer cells. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 7, pp. 54–59, July, 1994     

Na+/H+ countertransport and calcium exchange in peripheral blood cells of healthy subjects and patients with chronic heart failure

A comparative study is performed of Na⁺/H⁺ exchange and Ca²⁻ mobilization in erythrocytes and platelets of patients with stage I–II chronic heart failure caused by dilative cardiomyopathy and ischemic heart disease. A significant rise in the Na⁺/H⁺ exchange rate is found in the cells of chronic heart failure patients, which correlates with an elevated erythrocyte and platelet concentration of Ca²⁺ and an increased “calcium” response of platelets to inductors. The findings testify to a certain functional relationship between various cation-transporting cellular systems whose change in properties upon chronic heart failure can play an important pathogenic role. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 12, pp. 572–575, December, 1994     

Changed count of CD4+ T-lymphocytes in the bone marrow of aggressive CBA mice

Enhanced immune response of aggressive CBA mice after 10 daily confrontations in sensory contact on day 4 after immunization with sheep red blood cells (5×10⁸) is paralleled by an increase in the count of CD4⁺ T-cells in the bone marrow. Aggressive behavior, weight of the spleen, and count of CD4⁺ T-helpers in the bone marrow (which is increased only in aggressors with a history of at least 3 victories) are correlated. The effect of aggressive behavior on immunity can be caused by changes of the neurochemical status of the brain and determined by an increase in the CD4⁺ T-helper count. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 124, No. 11, pp. 544–546, November, 1997     

Oridonin promotes CD4+/CD25+ Treg differentiation, modulates Th1/Th2 balance and induces HO-1 in rat splenic lymphocytes

Objective and design: Oridonin is an ent-kaurene diterpenoid extracted from Isodon Serra, and we have previously demonstrated its immunosuppressive effect. Our goal was to study how Oridonin impacts CD4⁺/CD25⁺ regulatory T cells (Tregs) and Th1/Th2 balance, as well as its effect on the anti-inflammatory target HO-1. Material: Splenic lymphocytes were prepared from male 6–8-week-old SD rats. Treatment: Cells were cultured in four groups as Oridonin-L (Oridonin 12.5 μmol/l), Oridonin-H (Oridonin 25 μmol/l), Cobalt protoporphyrin (Copp 50 μmol/l) and control (DMSO) with stimulation of ConA (5 μg/ml) for 48 h or with no stimulation for 12 h. Method: We set up a model of Th1 polarization in vitro using ConA stimulation; ratios of CD4⁺/CD25⁺ Tregs (confirmed by the expression of Foxp3) were measured by flow cytometry, and levels of IL-2, IFN-γ, TGF-β and IL-10 were measured by ELISA. In addition, HO-1 expression was measured without stimulation with ConA by RT-PCR and Western blotting, and HO-1 level in vitro was then measured by enzyme activity assay. p<0.05 (t-test) was taken as the level of statistical significance. Result: Oridonin promoted differentiation towards CD4⁺/CD25⁺ Tregs, inhibited IL-2 and IFN-γ but induced TGF-β and IL-10, thus rectifed the Th1 polarization. Moreover, Oridonin induced the expression of HO-1 mRNA and protein, and HO-1 activity in vitro was enhanced accordingly. Conclusion: The results suggest that Oridonin has a distinct effect on promoting CD4⁺/CD25⁺ Treg differentiation and modulating Th1/Th2 balance, and this effect may be achieved via inducing the anti-inflammatory target HO-1. Received 16 October 2007; accepted by G. Wallace 26 November 2007     

Inhibition of mitogen-stimulated proliferation of human lymphocytesin vitro by synthetic peptide fragments of α-2 interferon

Human α-2 interferon and peptides representing parts of the amino-acid sequence 124–138 of the IF molecule inhibit the proliferative response of peripheral blood lymphocytes from healthy donorsin vitro induced by ConA. It is shown that neither interferon α-2 nor biologically active peptides change the level of interleukin-2 ConA-induced production by human blood mononuclears. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 117, N ^o 1, pp. 72–74, January, 1994 Presented by I. P. Ashmarin, Member of the Russian Academy of Medical Sciences     

Cytokine expression in CD4+ cells exposed to the monocyte locomotion inhibitory factor produced by Entamoeba histolytica

Entamoeba histolytica produces monocyte locomotion inhibitory factor (MLIF), a pentapeptide with in vitro and in vivo anti-inflammatory properties. MLIF may interfere with leukocyte migration, disturbing the balance of pro- and anti-inflammatory cytokines secreted by CD4⁺ T lymphocytes. We evaluated the effect of MLIF on expression of pro- and anti-inflammatory cytokines in human CD4⁺ T lymphocytes. Regulatory cytokines [interleukin-1 beta (IL-1β), IL-2, interferon gamma (IFN-γ), IL-5, IL-6, and IL-10] were studied by enzyme-linked immunosorbent assay method in CD4⁺-cell supernatant fluids. Proinflammatory cytokines were produced per se by MLIF (IL-1β, IL-2, and IFN-γ) and also anti-inflammatory cytokines (IL-5, IL-6, and IL-10) with 1-phorbol-12 myristate-13 acetate + MLIF; the IL-1β, IFN-γ, IL-5 and IL-6 production was inhibited but not that of IL-10 which disclosed increase in its expression. MLIF disturbs the pro- and anti-inflammatory balance, and it induces inhibition of IL-1β (principal proinflammatory cytokine) and increases IL-10 (prototype of an anti-inflammatory cytokine).     

Blast transformation of lymphocytes and the activity of natural killer cells in the presence of γ-globulinin vitro

The cytotoxic activity of natural killer cells against³H-uridine-labeled target cells (human erythromyeloleukosis cells K-562) and the intensity of spontaneous blast transformation are studiedin vitro in the presence of human serum γ-globulin. It is shown that spontaneous blast transformation is 49–51% due to the presence of aggregated γ-globulin, while the aggregate-free γ-globulin fraction does not induce this reaction. The cytotoxic activity of natural killer cellsin vitro declines in the presence of native γ-globulin, which is related to the influence of aggregated γ-globulin, the intensity of whose formation may increase upon a manyfold decrease in the γ-globulin content of the preparation. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 12, pp. 625–630, December, 1994     

A study of the mechanism of action of befol on Ca2+ metabolism in cardiomyocytes using a FURA-2 fluorescent probe

The dynamics of the Ca-response of cardiomyocytes is studied and the efficiency of befol, verapamil, and amiodarone is compared using various experimental models of stimulation of [Ca²⁺]_i. Befol (1–5 μM) is shown to inhibit the caffeine-and strophanthin G-induced rise of [Ca²⁺]_i. Unlike verapamil and amiodarone, befol exhibits no Ca-blocking activity in modeled K-depolarization. It is concluded that the cardiotropic effect of befol is mediated through its primary action on Na⁺/Ca²⁺ exchange in cardiomyocytes, while the cardioplegic effect of verapamil and amiodarone is due to their ability to block the slow Ca²⁺ inward current. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 121, N ^o 3, pp. 288–291, March, 1996     

Effect of recombinant human erythropoietin on the rate of Na+,H+-exchange in erythrocytes from patients with chronic renal failure on hemodialysis

The study explores the effect of recombinant human erythropoietin and the rate of Na⁺,H⁺-exchange in erythrocytes from patients with chronic renal failure undergoing hemodialysis. The rate of Na⁺,H⁺-exchange in erythrocytes from patients was higher than in the control and remained unchanged after 24 months of treatment with erythropoietin. Therapy with recombinant human erythropoietin does not normalize the Na⁺,H⁺-exchange mechanism. It is concluded that factors underlying disturbances of ion transport in erythrocytes from uremic patients cannot be corrected with erythropoietin. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 124, No. 12, pp. 613–615, December, 1997     

Absence of cytotoxic molecules in CD8- and/or CD56-positive adult T-cell leukaemia/lymphoma

Adult T-cell leukaemia/lymphoma (ATLL) cells usually exhibit a CD4⁺ (helper/inducer) phenotype (CD4⁺/8^–/56^–), and only a minority of tumours express the CD8 (cytotoxic/suppressor) or CD56 (natural killer [NK]-associated) antigens. TIA-1 is a cytotoxic granule-associated protein expressed in NK cells and cytotoxic T lymphocytes (CTLs). Granzyme B, perforin and Fas ligand (FasL) are also expressed in activated CTLs and NK cells. To clarify the cytotoxic potential of ATLL cells, immunohistochemistry was performed in CD8⁺ and/or CD56⁺ ATLL cells, using anti-TIA-1, anti-granzyme B, anti-perforin and anti-FasL antibodies. We studied nine cases of CD8⁺ and/or CD56+ ATLL, all of which exhibited monoclonal integration of human T-cell leukaemia virus type 1 (HTLV-1) proviral DNA. Four cases exhibited a CD8⁺/CD56^– phenotype, four others had a CD8^–/CD56⁺ phenotype, and one was CD8⁺/CD56⁺. All but one case also expressed the surface antigens CD3, TCR αβ, and CD4. Expression of granzyme B and TIA-1 were demonstrated in three and two cases, respectively, but none expressed perforin or FasL. In the control study, 10 cases with typical CD3⁺/4⁺/8^–/56^– ATLL demonstrated no expression of those cytotoxic-associated proteins. Our findings suggest that CD8 and/or CD56 positivity probably confer(s) no cytotoxic function on ATLL cells, and it is possible that CD8 and CD56 may be simply aberrant surface markers in ATLL. Received: 20 January 1999 / Accepted: 13 April 1999     

Effect of ionenes on ion permeability of erythrocyte membranes

Ionophore activity of ionenes, like that of polymyxins, is shown to be potentiated by long-chain antiions. Rapid stoichiometric anion equilibration in isoosmotic buffered sugar medium revealed that the amplitude of Cl⁻/OH⁻ exchange in erythrocytes induced by an ionene-detergent complex gradually drops with the decrease of both the charge density and the number of monomeric moieties in the polymeric chain. This is also paralleled by passive K⁺ leakage from the cells. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N ^o 7, pp. 44–46, July, 1994