Plasma γ-globulin fraction proteins, copper and zinc cations, and metal complexes formed by them with human serum γ-globulin induce the production of early (24-h incubation) IL-1β by human blood cells. The protein modified by Zn cations 1.2 times more actively (p<0.01) induced early IL-1β than the control γ-globulin, while γ-globulin metal complex with copper was 1.4 times less active (p<0.1) than the control protein. The regularities of induction changed over the course of 48-h incubation: zinc cations chelated by γ-globulin fraction protein reduced, while copper cations stimulated the realization of the protein induction potential in the metal complex.

As it was established in our previous studies, the proteins of human serum γ-globulin fraction could interact with copper or zinc ions distributed in the periglobular space, form metal complexes, and become able to perform effector functions differing due to the conformational shifts from those mediated by them in native conformation of their Fc regions. In the present work we have evaluated ability of the γ-globulin metal complexes formed with copper or zinc ions in the conditions like to the physiological ones to induce production or to regulate induction in the culture of freshly isolated human peripheral blood cells (PBC) of granulocyte (G) and granulocyte-macrophage (GM) colony-stimulating factors (CSF) as well as of vascular endothelial growth factor (VEGF). The γ-globulin metal complexes formed with both copper and zinc ions were found to similarly reduce production of GM-CSF, G-CSF, and VEGF induced in normal human PBC cultures by the control γ-globulins or by copper and zinc ions used alone. In context of theory and practice of inflammation the properties of the γ-globulin metal complexes might impact the basic knowledge in search of novel approaches to anti-inflammatory drugs development.

Interferon-alpha was detected in IFN pool produced by human leukocytes in the presence of gamma-globulin fraction proteins, copper and zinc cations, and metal-modified gamma-globulins. The cytokine appeared in culture medium at early terms (24 h) of incubation, is characterized by acid resistance, and is neutralized by antibodies to IFN-alpha. The content of IFN-alpha in supernatants of induced leukocytes reached 60-90 pg/ml and correlated with antiviral activity of the samples. Zinc bound to human serum gamma-globulin attenuated and copper stimulated the realization of IFN-inducing characteristics of the protein at early terms of incubation.

Определение IL-1β, вырабатываемОгО клетками крОви челОвека в присутствии белкОв γ-глОбулинОвОй фракции и их металлОкОмплексОв Чекнев С.Б., Ефремова И.Е., Писковская Л.С., Юшковец Е.Н., Бабаянц А.А. Лаборатория межклеточных взаимодействий НИИ эпидемиологии и микробиологии им. Н.Ф. Гамалеи Минздравсоцразвития РоссииРезюме. В работе показано, что в общем пуле цитокинов, вырабатываемых клетками крови челове-ка в присутствии белков γ-глобулиновой фракции, их металлокомплексов с медью и цинком, а также катионов меди и цинка, примененных изолированно, содержится до 148,0±14,9 пг/мл интерлейкина-1β (IL-1β). Трансформированные связыванием катионов цинка и меди γ-глобулины индуцируют в 2,0 раза (р = 0,1) и в 1,4 раза соответственно меньше IL-1β, чем контрольные белки. При этом метал-локомплекс γ-глобулина с медью оказывается в 2,7 раза (р < 0,01) более активным индуктором IL-1β, чем белок, связавший катионы цинка. В контрольных на связанный металл дозах катионы цинка, примененные изолированно, практически не индуцируют выработку IL-1β, катионы меди в 1,6 раза (р < 0,05) более активны, чем связавший их γ-глобулин, и в 1,45 раза (р < 0,1) эффективнее ФГА. Обсуждается участие катионов меди и цинка, хелатируемых из микроокружения антителами и транс-формирующих Fc регионы молекул антител, в регуляции выработки IL-1β клетками крови человека, индуцированными посредством активации их Fc рецепторов.Ключевые слова: γ-глобулин, металлокомплексы, индукция, выработка. Cheknev S.B., Efremova I.E., Piskovskaya L.S., Yushkovets E.N., Babajanz A.A.Detection of iL-1ββ proDuceD by human bLooD ceLLs in presence of proteins from the γ-gLobuLin fraction anD their compLexes with metaLs abstract. Present study has shown that a total cytokine pool produced by human blood cells in presence of proteins from the γ-globulin fraction, or their complexes with copper and zinc, as well as with pure copper and zinc cations, contains up to 148.0±14.9 pg interleukin-1β (IL-1β) per mL. The γ-globulins modified by copper or zinc binding induce, resp., 2.0 and 1.4-fold less IL-1β than intact proteins (significant by p = 0.1). Meanwhile, the copper-γ-globulin proved to be 2.7-fold more active (р < 0.01) IL-1β inducer, as a zinc-protein complex. Pure zinc ions did not induce IL-1β production, whereas induction rates with pure copper ions are 1.6-fold higher (p < 0.05) than with γ-globulin, and 1.45-fold more efficient that PHA stimulation (p < 0.1).A potential participation of copper and zinc cations bound to antibodies from the microenvironment and, thus, transforming Fc regions of antibody molecules, is discussed with regard of regulation of IL-1β production by human blood cells induced by activation of their Fc receptors.

Plasma γ-globulin fraction proteins, copper and zinc cations, and metal complexes of these cations and human serum γ-globulin induce the production of TNF-α by human blood cells. The protein modified by zinc cations is by 1.4-1.7 times more potent (p<0.001-0.01) than control γ-globulin in inducing the production of TNF-α, while metal complex formed by γ-globulin and copper is by 1.9-2.2 times more potent that the control protein (p<0.001). Under conditions of experimental induction, TNF-α is produced as a typical early response cytokine. During long-term incubation, copper cations lose the ability to induce TNF-α production, while in combination with γ-globulin these cations produce a synergistic effect with the control protein.