Autologous Enhancement by Interferon of Natural Killer Activity of Human Peripheral Blood Lymphocytes

The in vitro action of interferon (IFN)-α and IFN-γ from six healthy donors and ten patients with multiple sclerosis (MS) on natural killer (INK) activity of peripheral blood lymphocytes (PBL) was studied in an autologous system. The NK activity of PBL was detected by a cytotoxic test using 3H-uridine human erythromyeloblast K562 cells. Autologous IFN-α and IFN-γ did not augment NK activity of PBL from healthy donors in vitro, whereas in samples from MS patients the IFNs strongly stimulated NK cell cytotoxic function. This stimulation suggests the existence of an inhibitor of regulatory IFN action, that is produced in healthy donors simultaneously with IFN in response to IFN induction, but which is lacking in commercial IFN preparations. The factor-containing supernatants from healthy donors reduced the stimulatory action of autologous IFNs in patients with MS almost until complete blockade. Because this inhibitor was absent in patients with MS, deficiency of an inhibitor of IFN regulatory action in MS could open the way to treatment of this compartment of the immune system

IN MAMORY OF ALEXANDER KULBERG

Памяти Александра Яковлевича Кульберг

EXPRESSION OF THE CYTOKINE GENES INDUCED BY HUMAN SERUM γ-GLOBULIN AND BY ITS METAL COMPLEXES WITH ZINC

Synthesis of mRNAs for the key immunoactive cytokines was studied in mononuclear cells (MNC) from human peripheral blood, using reverse transcription and polymerase chain reaction, after incubation of isolated cells with human serum γ-globulin, zinc ions, or zinc-γ-globulin metal complexes. A constitutive presence of interferon-α (IFNα) and interleukin-18 (IL-18) mRNA was shown in MNC of all of the subjects observed, whereas IFNγ, IL-4, or IL-8 mRNA synthesis was not detectable. Expression of IL-1β, IL-2, IL-6 и IL-10 was revealed in only a half of cases. It has been shown that pure γ-globulins did induce IFNγ gene expression and enhance transcriptional activity of IL-1β and IL-2 genes. Zinc cations did also promote transcription of IL-1β gene, along with reduced synthesis of IL-2 and IFNγ mRNA, compared to the γ-globulin. In presence of γ-globulin-zinc complexes, IL-1β mRNA was synthesized at the same level, as following induction with control γ-globulin, whereas IL-2 gene expression was reduced, and IFNγ mRNA synthesis was abolished. The results are discussed in terms of immune response polarization in presence of γ-globulins and their metal complexes with zinc

Protective effects of zinc ions towards <i>Staphylococcus aureus</i> bacteria exposed with antibiotics

Objective. The work was performed with the purpose to study susceptibility of S. aureus bacteria to the action of the standard spectrum of antibiotics in presence of zinc ions used on the disks with antibiotics or on the lawn of the bacterial culture preliminarily to antibiotics treatment.Materials and methods. Suspensions of S. aureus bacteria which contained 108 CFU/ ml were sown by the lawns into the standard Petri dishes coated with the supplemented Nutrient Agar. 30 min later the standard disks with antibiotics were passed on the surface of the lawn, and zinc sulfate was added by the drops of the volume of 5 μl on the surface of the disk. Then the dishes with bacterial cultures were incubated for 24 hrs at 37°C followed by measuring diameter of the area of culture growth inhibition. In some tests preliminarily to the disks with antibiotics passing the places of following disks application were treated with the zinc sulfate for 10 min at the room temperature.Results and discussion. In presence of 1.0 μg/ml of zinc ions on the disk with antibiotics protective action of the metal towards the bacteria was registered at 2.9 per cent observations. In presence of one, four or eight zinc ions per one molecule of antibiotic protective action was registered at 1.4-5.7 per cent observations. Treatment with zinc ions of the surfaces of lawns followed by the disks installation resulted in 27.3-45.5 per cent observations of reducing diameter of the area of bacterial growth inhibition.Conclusion. The treatment of the surface of the lawn of S. aureus bacteria with zinc ions cause protection of the bacteria from the following inhibitory antibiotics action

INHIBITION OF GROWTH OF BACTERIA IN STAPHYLOCOCCUS AUREUS AND PSEUDOMONAS AERUGINOSA CULTURES BY COPPER AND ZINC CATIONS, APPLIED AT PHYSIOLOGICAL CONCENTRATIONS

Aim. Evaluation of antibacterial effect of y-globulin fraction bound and free copper and zinc cations, applied in cultures of S. aureus and P. aeruginosa at physiological (micromolar) concentrations. Materials and methods. Day cultures of S. aureus or P. aeruginosa were transferred from agar to physiological solution, and cell suspension was prepared, containing approximately 103 - 104 CFU/ml. Samples of metal-complexes of y-globulin with copper and zinc cations (30 and 45 pg/ml), control y-globulins (30 and 45 pg/ml) and salt solutions of copper and zinc, cation content in those corresponded to the quantity of the metal, that had bound with the protein at the stage of metal-complex obtaining (75 ng/ml), were introduced into the suspension. The suspensions were incubated at 37°C for 6 hours, sampling and CFU count according to the accepted micromethod was carried out every 2 hours. By the end of incubation (6 hours of observation) the suspensions were transferred into nutrient broth, thermostated for 1 day at 37°C, transparency of the nutrient broth compared with control (sterile) was evaluated afterwards. Results. Toxic effect of copper and zinc cations is detected starting from the 3rd hour of observation in S. aureus culture. Viable bacteria are absent in the culture with zinc after 6 hours, with copper - after 4 hours of incubation, y-globulin, that had bound copper cations, reduces the quantity of viable cells compared with control protein by 11.9 - 33.0% (

CONFORMATION CHANGES OF HUMAN SERUM γ–GLOBULIN IN THE PRESENCE OF ZINC IONS

Abstract. Conformational changes of human serum γ–globulin during interaction with the zinc ions were studied in a solution. It has been shown that the presence of zinc in over its physiological concentrations led to increase in optical density across the whole spectrum of γ–globulin ultraviolet absorption. On the contrary, hypochromia in the spectrum was registered after interaction of the protein with zinc used in subphisiological concentrations. Possible role of divalent metal cations in changes in conformation of the blood serum γ–globulins, and thereby in regulation of their effector functions was discussed. (Med. Immunol., 2005, vol.7, № 4, pp. 375–380

EVALUATION OF IL-6 PRODUCTION BY HUMAN BLOOD CELLS INCUBATED WITH METAL COMPLEXES OF Γ-GLOBULIN

Abstract. This study has shown that that a common cytokine pool induced in cultured human peripheral blood cells (PBC) supplied by either γ-globulin fraction proteins, copper or zinc cations, or appropriate metal complexes, contains detectable amounts of IL-6 (0.39+0.14 to 2.04+0.16 ng/ml). γ-globulin complexes with zinc or copper ions are able to induce production of IL-6 in amounts differing from those induced by control proteins, or copper and zinc ions used alone. IL-6 production by PBC in presence of γ-globulin/zinc complexes was 1.5 to 2.5-fold higher than with control proteins or single zinc ions at 24, 48 and 72 hrs of observation, whereas IL-6 production by PBC in presence of γ-globulin complexes with copper was higher than with appropriate control proteins or copper ions used alone for 48 and 72 hrs of cell incubation (resp., 1.7 to 2.4 and 2.8 to 4.6-fold increase). Possible role of copper and zinc ions chelated by γ-globulins from the microenvironment and modifying the Fc regions of antibodies, is considered a potential regulatory factor of IL-6 production by human PBC

Inhibition of hemolytic activity of Streptococcus pyogenes in mechanisms of antibacterial action of zinc ions

Aim. The work was performed with the purpose to study a hemolytic activity in the culture of S.pyogenes under the inhibitory action of millimolar concentrations of zinc ions.Materials and methods. Suspensions of S.pyogenes bacteria which contained 108 CFU/ml were sown by the lawns into the standard Petri dishes coated with the supplemented Blood Nutrient Agar. 30 min later the salt solutions of zinc or copper which contained the metals at the concentrations ranged between 5 x 10-3 M to 5 x 10-1 M were added by the 5 μl drops on the surfaces of the lawns with use of 36-channel stamp replicator. Then the dishes with bacterial cultures were incubated for 24 hrs at 37°C followed by measuring diameter of the area of culture growth inhibition and of the area of inhibition of hemolysis. The study was performed with use of controls towards measuring the state of bacterial cells obtained from different zones of the areas.Results. In presence of the zinc ions concentrations ranged between 50 to 500 mM the area of the growth inhibition of S.pyogenes was surrounded on the lawn of the bacterial culture by the area of the inhibition of hemolysis where the growth inhibition of S.pyogenes was not registered. Copper ions did not form such an area of the hemolysis inhibition.Conclusion. Inhibitory action of zinc ions on the hemolytic S.pyogenes activity in the culture seems to be specific and reversible, and is discussed in a context of the antivirulent zinc ions properties