Myelin basic protein charge isomers change macrophage polarization

Elene Tsitsilashvili,1 Maia Sepashvili,1,2 Marika Chikviladze,1 Lali Shanshiashvili,1,2,* David Mikeladze1,2,* 1Institute of Chemical Biology, Ilia State University, Tbilisi, Georgia; 2Department of Biochemistry, I. Beritashvili Center of Experimental Biomedicine, Tbilisi, Georgia *These authors con...

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Autores principales:	Tsitsilashvili E, Sepashvili M, Chikviladze M, Shanshiashvili L, Mikeladze D
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2019
Materias:	Arginase-1 iNOS HMGB1 RAGE Pathology RB1-214 Therapeutics. Pharmacology RM1-950
Acceso en línea:	https://doaj.org/article/f0342bb1666a4f6998a4bff783a7b4fa
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spelling	oai:doaj.org-article:f0342bb1666a4f6998a4bff783a7b4fa2021-12-02T01:48:38ZMyelin basic protein charge isomers change macrophage polarization1178-7031https://doaj.org/article/f0342bb1666a4f6998a4bff783a7b4fa2019-01-01T00:00:00Zhttps://www.dovepress.com/myelin-basic-protein-charge-isomers-change-macrophage-polarization-peer-reviewed-article-JIRhttps://doaj.org/toc/1178-7031Elene Tsitsilashvili,1 Maia Sepashvili,1,2 Marika Chikviladze,1 Lali Shanshiashvili,1,2,* David Mikeladze1,2,* 1Institute of Chemical Biology, Ilia State University, Tbilisi, Georgia; 2Department of Biochemistry, I. Beritashvili Center of Experimental Biomedicine, Tbilisi, Georgia *These authors contributed equally to this work Purpose: During a neuronal injury, a variety of immune cells infiltrate into the local microenvironment at the demyelination site. After the destruction of the intact myelin sheath, its major constituent myelin basic protein (MBP) dissociates from the plasma membrane and acts as a free ligand on the infiltrated immune cells. MBP exhibits charge microheterogeneity as a result of post-translational modifications, but the effect of various isomers of MBP on the activity of macrophages is not known.Materials and methods: MBP was isolated and puriﬁed from bovine brain white matter. RAW 264.7 macrophages were cultured in DMEM supplemented with heat-inactivated fetal bovine serum. For evaluation of macrophage polarization following treatment of RAW 264.7 cells with MBP charge isomers, inducible nitric oxide synthase (iNOS) expression (M1 phenotype marker) and arginase-1 expression (M2 phenotype marker) were determined in cell lysates by ELISA. To assess Rac activity, G-LISA Rac Activation Assay system was used. The expression of receptor for advanced glycation end-products (RAGE) and high mobility group box 1 (HMGB1) protein were assayed by Western blot analysis.Results: Our results have shown that minimally modified C1 component of MBP increases the expression of arginase-1 in cells, decreases the expression of iNOS, does not change the secretion of HMGB1 protein, but significantly elevates surface expression of RAGE, and in parallel, increases the activity of small GTPase Rac. On the other hand, highly modified deiminated isomer C8-MBP increases the secretion of HMGB1 protein but does not change the expression of arginase-1 or the content of RAGE.Conclusion: These data indicate that deiminated C8 isomer of MBP tends to polarize RAW macrophages into M1 phenotypes, whereas C1 enhances the activity of M2 phenotype markers. Keywords: Arginase-1, iNOS, HMGB1, RAGETsitsilashvili ESepashvili MChikviladze MShanshiashvili LMikeladze DDove Medical PressarticleArginase-1iNOSHMGB1RAGEPathologyRB1-214Therapeutics. PharmacologyRM1-950ENJournal of Inflammation Research, Vol Volume 12, Pp 25-33 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Arginase-1 iNOS HMGB1 RAGE Pathology RB1-214 Therapeutics. Pharmacology RM1-950
spellingShingle	Arginase-1 iNOS HMGB1 RAGE Pathology RB1-214 Therapeutics. Pharmacology RM1-950 Tsitsilashvili E Sepashvili M Chikviladze M Shanshiashvili L Mikeladze D Myelin basic protein charge isomers change macrophage polarization
description	Elene Tsitsilashvili,1 Maia Sepashvili,1,2 Marika Chikviladze,1 Lali Shanshiashvili,1,2,* David Mikeladze1,2,* 1Institute of Chemical Biology, Ilia State University, Tbilisi, Georgia; 2Department of Biochemistry, I. Beritashvili Center of Experimental Biomedicine, Tbilisi, Georgia *These authors contributed equally to this work Purpose: During a neuronal injury, a variety of immune cells infiltrate into the local microenvironment at the demyelination site. After the destruction of the intact myelin sheath, its major constituent myelin basic protein (MBP) dissociates from the plasma membrane and acts as a free ligand on the infiltrated immune cells. MBP exhibits charge microheterogeneity as a result of post-translational modifications, but the effect of various isomers of MBP on the activity of macrophages is not known.Materials and methods: MBP was isolated and puriﬁed from bovine brain white matter. RAW 264.7 macrophages were cultured in DMEM supplemented with heat-inactivated fetal bovine serum. For evaluation of macrophage polarization following treatment of RAW 264.7 cells with MBP charge isomers, inducible nitric oxide synthase (iNOS) expression (M1 phenotype marker) and arginase-1 expression (M2 phenotype marker) were determined in cell lysates by ELISA. To assess Rac activity, G-LISA Rac Activation Assay system was used. The expression of receptor for advanced glycation end-products (RAGE) and high mobility group box 1 (HMGB1) protein were assayed by Western blot analysis.Results: Our results have shown that minimally modified C1 component of MBP increases the expression of arginase-1 in cells, decreases the expression of iNOS, does not change the secretion of HMGB1 protein, but significantly elevates surface expression of RAGE, and in parallel, increases the activity of small GTPase Rac. On the other hand, highly modified deiminated isomer C8-MBP increases the secretion of HMGB1 protein but does not change the expression of arginase-1 or the content of RAGE.Conclusion: These data indicate that deiminated C8 isomer of MBP tends to polarize RAW macrophages into M1 phenotypes, whereas C1 enhances the activity of M2 phenotype markers. Keywords: Arginase-1, iNOS, HMGB1, RAGE
format	article
author	Tsitsilashvili E Sepashvili M Chikviladze M Shanshiashvili L Mikeladze D
author_facet	Tsitsilashvili E Sepashvili M Chikviladze M Shanshiashvili L Mikeladze D
author_sort	Tsitsilashvili E
title	Myelin basic protein charge isomers change macrophage polarization
title_short	Myelin basic protein charge isomers change macrophage polarization
title_full	Myelin basic protein charge isomers change macrophage polarization
title_fullStr	Myelin basic protein charge isomers change macrophage polarization
title_full_unstemmed	Myelin basic protein charge isomers change macrophage polarization
title_sort	myelin basic protein charge isomers change macrophage polarization
publisher	Dove Medical Press
publishDate	2019
url	https://doaj.org/article/f0342bb1666a4f6998a4bff783a7b4fa
work_keys_str_mv	AT tsitsilashvilie myelinbasicproteinchargeisomerschangemacrophagepolarization AT sepashvilim myelinbasicproteinchargeisomerschangemacrophagepolarization AT chikviladzem myelinbasicproteinchargeisomerschangemacrophagepolarization AT shanshiashvilil myelinbasicproteinchargeisomerschangemacrophagepolarization AT mikeladzed myelinbasicproteinchargeisomerschangemacrophagepolarization
_version_	1718402932055998464

Myelin basic protein charge isomers change macrophage polarization

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