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Respiratory burst of neutrophil acting in vitro against melanoma F10

It is assumed that oxidative damage caused by oxygen free radicals from activated neutrophils may contributes to pathology of tumor. The present study is focused on the in vivo and in vitro effects of neutrophils when challenged with melanoma B16F10. In this study we were evaluating the respiratory burst of neutrophils by chemiluminescence. Male C57Bl/Ly5 mice were used. There were four different groups of animals: control mice, Sephadex G-200 subcutaneously bearing injectecd, melanoma bearing mice and melanoma bearing mice injected s.c. with Sephadex. Neutrophils from blood and neutrophils from papula (formed at the side were Sephadex G-200 was injected) were analyzed. The respiratory burst was evaluated as singlet oxygen (1O2) release, by luminol amplified chemiluminescence. The data were obtained from resting or PMA activated neutrophils and from PMA activated peripheral blood cells. Peripheral blood leukocyte counts, leukocyte subsets in peripheral blood and the survival of animals were evaluated, too. Our results showed significant decrease of blood leukocyte count on the second day after Sephadex injection and tumor transplantation in both groups. Sephadex treated melanoma bearing mice had lower survival rate than those bearing melanoma only. Neutrophils from melanoma bearing mice showed high production of singlet oxygen while neutrophils from mice without melanoma did not recognize B16F10 cells in vitro and had lower 1O2 production. On the 2nd day PMA activated blood cells had low capacity in 1O2 production in all groups except in control without tumor and Sephadex. This was probably due to the interactions of granulocytes in vivo with melanoma cells and/or with Sephadex particles. Blood neutrophils of mice without melanoma showed higher chemiluminescence on 9th day suggesting that the tumor antigens could provoke neutrophils to antitumorous-response only in an early period of tumor growth.

respiratory burst; neutrophil; melanoma F10

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