Open AccessCelestine blue B as a sensor for hypochlorous acid and HOCL-modified proteins registration
Author(s) -
Veronika E. Lutsenko,
Луценко Вероника Евгеньевна,
Daria V. Grigorieva,
Григорьева Дарья Владимировна,
Irina V. Gorudko,
Горудко Ирина Владимировна,
С. Н. Черенкевич,
Черенкевич Сергей Николаевич,
Н. П. Горбунов,
Горбунов Николай Петрович,
Valeria A. Kostevich,
Костевич Валерия Александровна,
О. М. Панасенко,
Панасенко Олег Михайлович,
A. V. Sokolov,
Соколов Алексей Викторович
Publication year - 2019
Publication title -
medicinskij akademičeskij žurnal
Language(s) - English
Resource type - Journals
eISSN - 2687-1378
pISSN - 1608-4101
DOI - 10.17816/maj19263-71
Subject(s) - hypochlorous acid , myeloperoxidase , chemistry , biochemistry , hydrazide , in vitro , immunology , organic chemistry , inflammation , biology
Objective — the study of hypochlorous acid (HOCl) and its derivatives production, which catalyzed by human neutrophil myeloperoxidase, using “turn-on” fluorescent sensor — celestine blue B.
Materials and methods. Neutrophils were isolated from the venous blood of healthy donors. Phorbol 12-myristate 13-acetate, N-formyl-methionyl-leucyl-phenylalanine, plant lectins, HOCl-modified proteins were used as agonists. N-acetylcysteine, 4-aminobenzoic acid hydrazide, isoniazid and ceruloplasmin were used as regulators of neutrophil myeloperoxidase activity and/or HOCl scavengers.
Results. Using a wide range of agonists and inhibitors, it has been shown that celestine blue B is oxidized in vitro by HOCl and its derivatives as a result of neutrophil myeloperoxidase activity. The oxidation of celestine blue B by HOCl-modified human serum albumin (HSA-Cl) and inhibition of this process by monoclonal antibody against HSA-Cl (IgM class) was also found.
Conclusion. Based on the developed method using celestine blue B, it is possible to conduct a sensitive analysis for the presence of HOCl-modified proteins (chloramines, etc.), to investigate the effect of various agonists and drugs on myeloperoxidase activity and exocytosis from the neutrophil granules.