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There is an everincreasing demand for lupin seed protein in human nutrition. However, their usage as a food ingredient can be limited by their influence on the immune system. The aim of the study was to analyse, by means of flow cytometry, the potential of lupin seed globulins to induce the respiratory burst in human neutrophils. The results proved the potential of lupin globulins to induce the increased ROS production in human neutrophils. The increased ROS production by the cells treated with lupin globulins and, at the same time, stimulated with the PMA may suggest a synergistic effect of the globulins and the PMA.
Our project aimed to investigate the effects of mercury on the proliferation of human peripheral lymphocytes in vitro. The lymphocytes were isolated from the blood collected from healthy donors at Regionalne Centrum Krwiodawstwa i Krwiolecznictwa in Poznań, Poland. For the purpose of cell culture, the lymphocyte suspension (25・10⁴ cells/ml) in Eagle’s medium supplemented with 10% fetal calf serum was prepared. Phytohaemagglutinin-L (PHA-L) was used in a concentration of 2.5 mg/ml to stimulate cell proliferation. Mercuric chloride (HgCl₂) in four different concentrations (1 μM, 10 μM, 50 μM, 100 μM) and [3H]-thymidine were added after 48 hours of incubation and the cell culture was continued for the next 24 hours. The rate of lymphocyte proliferation was measured by [3H]-thymidine incorporation method with a liquid scintillation counter. Results indicate that higher concentrations of mercury (50 μM, 100 μM) inhibit the [3H]-thymidine incorporation of human peripheral lymphocytes in vitro. The incorporation was lower than the control sample by 65% at a concentration of 50 μM, while at a concentration of 100 μM it fell to virtually zero. Moreover, the phase of lymphocyte proliferation cycle affected by mercuric chloride was also investigated. For this purpose HgCl2 in 2 concentrations (10 μM, 50 μM) was added to the cell culture in 4 different timepoints: at the start of the cell culture and after 4, 24, and 48 hours of incubation. After 48 hours, [3H]-thymidine was added and the cell culture was continued for an additional 24 hours. The rate of cell proliferation was estimated by [3H]-thymidine incorporation using a liquid scintillation counter. The inhibition effect was observed in samples with metal added at the start of the cell culture and after 4 h of incubation, i.e. at the initial phase of the lymphocyte proliferation cycle.
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