Granule Associated DNase in T-Lymphocytes from Patients with Inflammatory Disease

Bera C. Скрипов¹, Cyrille A. VODOUNON^{1, 2,} , Irina D. RESHETNIKOVA³, Boris B. LEGBA³ and Zinaida I. ABRAMOVA¹

¹Department of Biochemistry, Institute of Fundamental Medicine and Biology, Kazan (Volga Region) Federal University, Kazan, Russian Federation

²Laboratoire de Biochimie et de Microbiologie, Ecole Normale Supérieure de Natitingou (ENS), Université de Parakou, Parakou-Bénin

³Clinic of allergic diseases of Kazan Research Institute of Epidemiology and Microbiology, Kazan, Russian Federation

Cite this paper:
Bera C. Скрипов, Cyrille A. VODOUNON, Irina D. RESHETNIKOVA, Boris B. LEGBA and Zinaida I. ABRAMOVA. Granule Associated DNase in T-Lymphocytes from Patients with Inflammatory Disease. American Journal of Biomedical Research. 2016; 4(4):87-93. doi: 10.12691/ajbr-4-4-2

Abstract

Recent data highlight the undeniable role of programmed cell death type I of lymphocytes in the pathogenesis of certain allergic diseases and autoimmune diseases such as Bronchial Asthma and hemorrhagic rectocolitis. But little data exist on the enzymatic activity of secretory granules associated with lymphocytes of patients suffering from these inflammatory diseases. The aim of the study was to characterize the activity of the DNase in the secretory granules of T lymphocytes isolated from peripheral blood of patients with Bronchial Asthma (n = 20) and Hemorrhagic Rectocolitis (n = 20). Thus, the secretory granules were isolated from lymphocytes by the ultracentrifugation method on percoll density gradient. The detection of the activity of the protein extracts was performed by zymography and electrophoresis method. The results reveal the presence of a protein extract with a molecular weight of 66 kDa both at the level of the granules of the lymphocytes of patients suffering from hemorrhagic rectocolitis considered as a classical autoimmune disease and in the granules of lymphocytes of patients with Bronchial Asthma. The study of physicochemical properties showed an increase in the enzymatic activity of DNase of secretory granules when 1 mM Ca²⁺ was added to the incubation medium at a pH = 7.5. On the other hand, the addition of 1 mM Zn^{2 +} causes the inhibition of enzyme activity. These results suggest that the enzymatic activity of DNase detected in the granule of lymphocytes of patients with Bronchial Asthma and hemorrhagic rectocolitis occurs not only in the fragmentation of double stranded DNA but could also play a role in the apoptotic process of these same T-lymphocytes. The study of the properties of this DNase in the inflammatory diseases could enable to use this protein as a marker for determining the severity of disease.

Keywords:
DNase secretory granules Bronchial Asthma Hemorrhagic Rectocolitis

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References:

[1]	Ярилин, А.А. Base of Immunology. – М.: Меdicine, 1999. 608 с.
[2]	Green, D.R. Ferguson, L. Zitvogel, G. Kroemer. Immunogenic and tolerogenic cell death. Nat Rev Immunol 2009; 9: 353-363.
[3]	Elmore, E. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007; 35: 495-516.
[4]	Rubinsztein, D.C, Williams A, S. Sarkar, et al. Novel targets for Huntington's disease in an mTOR-independent autophagy pathway. Nat Chem Biol 2008; 4 (5):295-305.
[5]	Косарев, В.В , Лотков B.C, Куклин A.C et al. Ингаляционные кортикостероиды в терапии бронхиальной астмы. Терапевт, арх 2000; 8 : 59-61.
[6]	Spinozzi, F, Benedictis F.M. Apoptosis, airway inflammation and anti-asthma therapy: from immunobiology to clinical application. Pediatr Allergy Immunol 2008; 19 (4): 287-295.
[7]	Vodounon AC, Ckibo Y, Aikou N, Akpona S, Abramova ZI, Kotchoni OS,Baba-Moussa L. Morphological and biochemical characteristics of apoptosis lymphocytes of peripheral blood in the pathogenesis of atopic bronchial asthma of light and serious severity. Trade Sci Inc (Res. Rev BioSci) 2012; 6(8): 210-20.
[8]	Pio, R, Gonzalez A, Lopez-Zabalza et al. Granule associated DNase in T4 and T8 lymphocytes from patients with autoimmune diseases. Biochimica et Biophysica Acta 1998; 1406: 51-61.
[9]	Parrish, J.Z, Deng X. Cuts can kill: the roles of apoptotic nucleases in cell death and animal development. Chromosoma 2006; 115: 89-97.
[10]	Patel D, Rubbi CP, Rickwood D. Separation of T and B lymphocytes from human peripheral blood mononuclear cells using density perturbation methods. Clin Chim Acta 1995; 240: 187-93.
[11]	Ulmer, A.J, Scholz W, Ernest M et al. Isolation and subfractionation of human peripheral blood mononuclear cells (PBMC) by density gradient centrifugation on Percoll. Immunobiology 1984;166: 238-50.
[12]	Boyum, A, Brincker-Fjerdingstad I et al. Separation of human lymphocytes from citrated blood by density gradient (NycoPrep) centrifugation: monocyte depletion depending upon activation of membrane potassium channels. Scand J Immunol 2002; 56: 76-84.
[13]	F.A. Khorsid F.A and S.S. Mushref S.S. In vitro Anticancer Agent I-Tissue Culture Study of Human Lung Cancer Cells A549 II-Tissue Culture Study of Mice Leukemia Cells L1210. International Journal of Cancer Research 2006; 2: 330-344.
[14]	Borregaard N, Heiple J.M, Simons E.R, Clark R.A. Subcellular localization of the b-cytochrome component of the human neutrophil microbicidal oxidase: Translocation during activation. J. Cell Biol 1983; 97: 52-61.
[15]	Podack ER, Konigsberg PJ. Cytolytic T cell granules. Isolation, structural, biochemical, and functional characterization. J Exp Med 1984; 160: 695-710.
[16]	Thiery, J.,Walch M, Jensen D.K, Jensen D. Isolation of Cytotoxic T Cell and NK granules and purification of their effector proteins. Current Protocols in Cell Biology 2010; 3:37(1)3.37.29.
[17]	Laemmli, U.K. Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature 1970; 227: 680-685.
[18]	Napirei, M, Jensen A, Eulitz D et al. Expression pattern of the deoxyribonuclease 1 gene: lessons from the Dnase 1 knockout mouse. Biochem J. 2004; 380(Pt 3): 929-337.
[19]	Urboniene, D, Sakalauskas R, Sitkauskiene B. Autoimmunity in pathogenesis of chronic obstructive pulmonary disease. Medicina (Kaunas) 2005;4 (13): 190-195.
[20]	Kopinski, P, Przybylsk G, Balicka-Slusarczyk et al. Apoptosis of alveolar lymphocytes in sarcoidosis and in control group is more frequentin smokers then in nonsmoking persons. Przegl Lek. 2006; 63: 841-847.
[21]	Vignola, A.M, Chanez P, . Chiappara et al. Evaluation of apoptosis of eosinophils, macrophages, and T lymphocytes in mucosal biopsy specimens of patients with asthma and chronic bronchitis. J Allergy Clin Imunol 1999; 103:563-573.
[22]	Busse, W.W, Lemanska R.F. Аsthma. NEJM 2001; 344:350-362.
[23]	Мамонтова, Т.В, Kaidathev I.P. Новые аспекты апоптоза мононуклеарных клеток в патогенезе атопической бронхиальной астмы. Аllergology 2005;4: 15-23.
[24]	Chau-Ching, L, Young L.H.Y et al. Lymphocyte-mediated cytolysis and disease. N. Engl. J. Med; 1996; 335: 1651-1659.