Expression Level of Membrane Markers CD5, CD19, and CD20 in B Cells after UV-Irradiation and Incubation in the Presence of Autologous Plasma.

The effect of UV-light (240-390 nm) in doses of 151 and 755 J/m2 on the expression of membrane markers CD5, CD19, CD20 in human peripheral blood B cells was studied by flow cytometry. In 24 h after exposure to UV light, we observed activation of processes accompanied by structural rearrangements of B-cell membranes leading to changes in the expression of receptor molecules: the content of of CD19 and CD20 increased due to activation of the synthesis of these proteins, while the content of CD5 decreased. The percentage of CD5+ cells decreased over 24 h after UV-irradiation of lymphocytes, while addition of autologous plasma to the incubation medium produced a photoprotective effect on CD5+ cells.

[The Influence of UV-Light on the Sub-Populational Composition and Expression of Membrane Markers of Lymphocytes of Donor Blood].

The influence of UV-light (240-390 nm) at dozes of 151 and 755 J/m2 on the content of membrane markers of lymphocytes using the method of flow cytometry was investigated. It was demonstrated that during incubation of UV-irradiated lymphocytes the change of their populational and sub-populational composition occurs. Expression of complexes of CD3, CD 19,.CD8, CD 16, CD25 and CD95 increased. This increase was caused mainly by de novo synthesis. UV-light had immunostimulating effect on CD8+ T-lymphocyte population. Together with the increase of cytotoxic cells and NK-cells, activation of lymphocytes (increased amount of CD25+ and CD95+ cells) took place. Amount of cells undergone apoptosis or necrosis increased proportionally to the dosage. These changes were more expressed during incubation of lymphocytes in nutrition medium without autological blood serum, e.g. under deficiency of growth factors and antioxidants.

[Apoptosis and necrosis of lymphocytes induced by UV-radiation in the presence of autological plasma].

The influence of UV-light (240-390 nm) in doses 151-3020 J/m2 on the nature of the death of lymphocytes cells of donor's blood (using markers of apoptotic and necrotic death of cells) and on the level of expression of the marker of apoptotic pre-preparation--CD95-receptor has been investigated. We have shown that UV-radiation increases expression of CD95-receptors which is caused mainly by de novo synthesis of the receptors. It has been revealed that during daily incubation of photo-modified lymphocytes (151 and 755 J/m2) without autological blood cell death occurs by receptor-involved apoptosis. Exposure to high doses of radiation (1510 and 3020 J/m2) causes massive necrotic death of immunocytes. The use of autologous blood plasma during incubation of photo-modified lymphocytes allows decreasing the number of both apoptotic and necrotic cells.

[Influence of UV-light on the profile of receptors of lymphocytes of donated blood].

The influence of UV-light (240-390 nm) at the dozes of 151 and 755 J/m2 on the maintenance of membrane markers of lymphocytes: CD3, CD19, CD4, CD8, CD25, CD95 using the method of flow cytometry was investigated. It was demonstrated that UV-irradiation of lymphocytes induced the decrease in the number of CD3, CD4 and CD19 markers together with a simultaneous increase of the amount of CD8, CD25 receptors during the daily incubation of cells in a nutrient medium without plasma. When autologous plasma is used during incubation of both native and UV-irradiated lymphocytes, the percentage of cells with the specified markers is not different from the corresponding values in the cells immediately after their purification. This finding points at the important role of antioxidants of the blood plasma and growth factors for the maintenance of the sub-population structure of lymphocytes. However, the level of expression of CD95, one of the markers of apoptosis, increases in the photomodified cells incubated both in the presence and in the absence of plasma. This increase was revealed to be caused mainly by the synthesis of the protein de novo under the influence of apoptosis-inducing factors.

[The protective effect of autologous plasma against the development of oxidative stress in UV-irradiated lymphocytes of peripheral blood of donors].

The influence of UV-light (240-390 nm) at the dozes of 151 and 755 J/m2 on the intensity of processes of the lipid peroxidation, activity of lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), cytochrome c oxidase and the level of the energy supply of donors' blood lymphocytes in the absence and presence of autologous plasma was investigated. It was shown that during the incubation of native and UV-irradiated lymphocytes, autologous plasma reduces the intensity of lipid peroxidation, thus protecting cells from oxidative stress. As a result, the endocellular level of ATP is restored in UV-irradiated lymphocytes (during the daily incubation), which reflects the intensification of the adaptive ability of cells in the presence of autologous plasma.

[Peculiarities of metabolism of UV-irradiated lymphocytes].

The influence of UV-light (240-390 nm) at a dose of 151 and 755 J/m2 on the functional properties of lymphocyte metabolism key enzymes from donors' human blood: lactate dehydrogenase (LDH), cytochrome c oxidase, succinate dehydrogenase (SDH), Ca2(+)-ATPase of plasma membranes has been investigated. It has been revealed that photoinactivation of enzymes immediately after UV-irradiation which leads to the decrease of the ATP content in lymphocytes is replaced by the increased activity of the enzymes under investigation during daily incubation of lymphocytes. As a result, the level of ATP in photo-modified lymphocytes does not differ from that in native cells before incubation. This indicates the normalization of biochemical processes in lymphocytes influenced by UV-light applied in autotransfusion of UV-irradiated blood.