A test system for assessment of interferon gamma bioactivity

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Abstract

FN-y is a cytokine of great clinical significance. The excessive or insufficient function of the signalling pathway triggered by IFN-y is a cause of various pathological states of the immune system and the body in general. At present, clinical practice uses quantitative methods of detection of this protein without taking into account its functional activity. Traditional tests for assessment of the biological activity of IFN-y are technologically complex and their results are poorly quantified. In the course of this work a model system was created representing a cell line with a reporter construct stably integrated into the genome controlled by a STAT-dependent promotor. Processing of the cells of this model system by IFN-y activates the STAT-1-signalling pathway, which causes expression of FFly luciferase gene. Luciferase activity is quantitatively assessed by detection of bioluminescence. Working concentration values (2.5 ng/ml) and time of performing the assay (8 h) were chosen. The effectiveness of using the model system for testing IFN-y bioactivity has been shown.

About the authors

Ekaterina P. Konyaeva

Dmitry Rogachev Federal Research Centre of Paediatric Haematology, Oncology and Immunology, Ministry of Health of the Russian Federation; Moscow Institute of Physics and Technology (State University)

Author for correspondence.
Email: konyaeva@phystech.edu
Russian Federation

Kseniya V. Kulikova

Dmitry Rogachev Federal Research Centre of Paediatric Haematology, Oncology and Immunology, Ministry of Health of the Russian Federation; Institute of Gene Biology, Russian Academy of Sciences; N.I.Pirogov Russian National Research Medical University

Email: kv.kulikova@gmail.com
Russian Federation

Marina I. Lukashina

Dmitry Rogachev Federal Research Centre of Paediatric Haematology, Oncology and Immunology, Ministry of Health of the Russian Federation; Institute of Gene Biology, Russian Academy of Sciences; N.I.Pirogov Russian National Research Medical University

Email: mlukashina@mail.ru
Russian Federation

Nikolay V. Gnuchev

Institute of Gene Biology, Russian Academy of Medical Sciences

Email: gnuchev@igb.ac.ru
Russian Federation

Georgiy P. Georgiev

Institute of Gene Biology, Russian Academy of Medical Sciences

Email: georgiev@igb.ac.ru
Russian Federation

Sergey S. Larin

Dmitry Rogachev Federal Research Centre of Paediatric Haematology, Oncology and Immunology, Ministry of Health of the Russian Federation; Institute of Gene Biology, Russian Academy of Medical Sciences

Email: sergei_larin@mail.ru
Russian Federation

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Copyright (c) 2016 Konyaeva E.P., Kulikova K.V., Lukashina M.I., Gnuchev N.V., Georgiev G.P., Larin S.S.

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This work is licensed under a Creative Commons Attribution 4.0 International License.