PHENOTYPIC CHARACTERISATION OF PERIPHERAL BLOOD CYTOTOXIC T LYMPHOCYTES: REGULATORY AND EFFECTOR MOLECULES

Cytotoxic T lymphocytes (CD3+CD8+, Tcyt) play a major role in protective immunity against intracellular pathogens and can eradicate malignant cells. As based on CD45RA and CD62L expression, the peripheral CD3+CD8+ blood lymphocytes were divided into "na ve" (N) cells, central memory (CM) a...

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Autores principales:	I. V. Kudryavtsev, A. G. Borisov, E. V. Vasilyeva, I. I. Krobinets, A. A. Savchenko, M. K. Serebriakova, A. Totolian Areg
Formato:	article
Lenguaje:	RU
Publicado:	SPb RAACI 2018
Materias:	flow cytometry multicolor immunophenotyping cytotoxic t cell subsets cd3+cd8+ maturation effector molecules inhibitory receptors Immunologic diseases. Allergy RC581-607
Acceso en línea:	https://doaj.org/article/bcfc3ee5581440e3a5839c244882d677
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spelling	oai:doaj.org-article:bcfc3ee5581440e3a5839c244882d6772021-11-18T08:03:47ZPHENOTYPIC CHARACTERISATION OF PERIPHERAL BLOOD CYTOTOXIC T LYMPHOCYTES: REGULATORY AND EFFECTOR MOLECULES1563-06252313-741X10.15789/1563-0625-2018-2-227-240https://doaj.org/article/bcfc3ee5581440e3a5839c244882d6772018-03-01T00:00:00Zhttps://www.mimmun.ru/mimmun/article/view/1485https://doaj.org/toc/1563-0625https://doaj.org/toc/2313-741XCytotoxic T lymphocytes (CD3+CD8+, Tcyt) play a major role in protective immunity against intracellular pathogens and can eradicate malignant cells. As based on CD45RA and CD62L expression, the peripheral CD3+CD8+ blood lymphocytes were divided into "na ve" (N) cells, central memory (CM) and effector memory (EM), as well as "terminally-differentiated" CD45RA-positive effector cells (TEMRA). Using multicolor flow cytometry, a co-expression of effector (perforin, granzyme B and CD57) and regulatory (CD27, CD28, CD244 (2B4), CD279 (PD-1) and KLRG1) molecules was studied on all these subsets. CD57 was expressed in 2.39±0.31% “na ve” and 5.45±0.91% of central memory Tcyt. Meanwhile, within EM and TEMRA Tcyt subset, its expression was identified on the cell membranes of 26.53±2.20% and 51.43±2.55% of cells, respectively. Cytolytic effector molecules (granzyme B and perforin) were detected in cytoplasmic granules of 4.22±0.36% and 5.30±0.43% of na ve Tcyt, respectively. For CM cells, these values were 10.09±1.17% and 24.90±3.10%, respectively. Dramatic increases of granzyme B and perforin expression were observed in the “EM → TEMRA” cell lineage, when the relative number of granzyme B-positive cells increased to 41.05±2.63% and 66.73±3.29%, respectively, while perforin was detected in 59.33±4.26% and 75.08±3.12% of cells, respectively. For regulatory molecules, CD244 and KLRG1, the similar dynamics were observed, their expression increased from “na ve” to late maturation stages, while the expression of two main costimulatory molecules – CD27 and CD28, decreased in the lineage N → CM → EM → TEMRA cells. The highest level of CD279 was observed in EM cells. It was shown that CD57-positive cells contain perforin and granzyme B in their cytoplasmic granules and lack CD28 expression. Furthermore, CD57 can be used as a surrogate marker for multicolor immunophenotyping to identify most mature effector cells containing cytolytic enzymes. Our results on the co-expression of all the beforementioned molecules suggest that the most mature CD45RA+CD62L– effector peripheral blood cytotoxic T cells express CD244 and CD57, lack costimulation molecules CD27 and of CD28, as well as inhibitory receptors KLRG1 and CD279.I. V. KudryavtsevA. G. BorisovE. V. VasilyevaI. I. KrobinetsA. A. SavchenkoM. K. SerebriakovaA. Totolian AregSPb RAACIarticleflow cytometrymulticolor immunophenotypingcytotoxic t cell subsetscd3+cd8+ maturationeffector moleculesinhibitory receptorsImmunologic diseases. AllergyRC581-607RUMedicinskaâ Immunologiâ, Vol 20, Iss 2, Pp 227-240 (2018)
institution	DOAJ
collection	DOAJ
language	RU
topic	flow cytometry multicolor immunophenotyping cytotoxic t cell subsets cd3+cd8+ maturation effector molecules inhibitory receptors Immunologic diseases. Allergy RC581-607
spellingShingle	flow cytometry multicolor immunophenotyping cytotoxic t cell subsets cd3+cd8+ maturation effector molecules inhibitory receptors Immunologic diseases. Allergy RC581-607 I. V. Kudryavtsev A. G. Borisov E. V. Vasilyeva I. I. Krobinets A. A. Savchenko M. K. Serebriakova A. Totolian Areg PHENOTYPIC CHARACTERISATION OF PERIPHERAL BLOOD CYTOTOXIC T LYMPHOCYTES: REGULATORY AND EFFECTOR MOLECULES
description	Cytotoxic T lymphocytes (CD3+CD8+, Tcyt) play a major role in protective immunity against intracellular pathogens and can eradicate malignant cells. As based on CD45RA and CD62L expression, the peripheral CD3+CD8+ blood lymphocytes were divided into "na ve" (N) cells, central memory (CM) and effector memory (EM), as well as "terminally-differentiated" CD45RA-positive effector cells (TEMRA). Using multicolor flow cytometry, a co-expression of effector (perforin, granzyme B and CD57) and regulatory (CD27, CD28, CD244 (2B4), CD279 (PD-1) and KLRG1) molecules was studied on all these subsets. CD57 was expressed in 2.39±0.31% “na ve” and 5.45±0.91% of central memory Tcyt. Meanwhile, within EM and TEMRA Tcyt subset, its expression was identified on the cell membranes of 26.53±2.20% and 51.43±2.55% of cells, respectively. Cytolytic effector molecules (granzyme B and perforin) were detected in cytoplasmic granules of 4.22±0.36% and 5.30±0.43% of na ve Tcyt, respectively. For CM cells, these values were 10.09±1.17% and 24.90±3.10%, respectively. Dramatic increases of granzyme B and perforin expression were observed in the “EM → TEMRA” cell lineage, when the relative number of granzyme B-positive cells increased to 41.05±2.63% and 66.73±3.29%, respectively, while perforin was detected in 59.33±4.26% and 75.08±3.12% of cells, respectively. For regulatory molecules, CD244 and KLRG1, the similar dynamics were observed, their expression increased from “na ve” to late maturation stages, while the expression of two main costimulatory molecules – CD27 and CD28, decreased in the lineage N → CM → EM → TEMRA cells. The highest level of CD279 was observed in EM cells. It was shown that CD57-positive cells contain perforin and granzyme B in their cytoplasmic granules and lack CD28 expression. Furthermore, CD57 can be used as a surrogate marker for multicolor immunophenotyping to identify most mature effector cells containing cytolytic enzymes. Our results on the co-expression of all the beforementioned molecules suggest that the most mature CD45RA+CD62L– effector peripheral blood cytotoxic T cells express CD244 and CD57, lack costimulation molecules CD27 and of CD28, as well as inhibitory receptors KLRG1 and CD279.
format	article
author	I. V. Kudryavtsev A. G. Borisov E. V. Vasilyeva I. I. Krobinets A. A. Savchenko M. K. Serebriakova A. Totolian Areg
author_facet	I. V. Kudryavtsev A. G. Borisov E. V. Vasilyeva I. I. Krobinets A. A. Savchenko M. K. Serebriakova A. Totolian Areg
author_sort	I. V. Kudryavtsev
title	PHENOTYPIC CHARACTERISATION OF PERIPHERAL BLOOD CYTOTOXIC T LYMPHOCYTES: REGULATORY AND EFFECTOR MOLECULES
title_short	PHENOTYPIC CHARACTERISATION OF PERIPHERAL BLOOD CYTOTOXIC T LYMPHOCYTES: REGULATORY AND EFFECTOR MOLECULES
title_full	PHENOTYPIC CHARACTERISATION OF PERIPHERAL BLOOD CYTOTOXIC T LYMPHOCYTES: REGULATORY AND EFFECTOR MOLECULES
title_fullStr	PHENOTYPIC CHARACTERISATION OF PERIPHERAL BLOOD CYTOTOXIC T LYMPHOCYTES: REGULATORY AND EFFECTOR MOLECULES
title_full_unstemmed	PHENOTYPIC CHARACTERISATION OF PERIPHERAL BLOOD CYTOTOXIC T LYMPHOCYTES: REGULATORY AND EFFECTOR MOLECULES
title_sort	phenotypic characterisation of peripheral blood cytotoxic t lymphocytes: regulatory and effector molecules
publisher	SPb RAACI
publishDate	2018
url	https://doaj.org/article/bcfc3ee5581440e3a5839c244882d677
work_keys_str_mv	AT ivkudryavtsev phenotypiccharacterisationofperipheralbloodcytotoxictlymphocytesregulatoryandeffectormolecules AT agborisov phenotypiccharacterisationofperipheralbloodcytotoxictlymphocytesregulatoryandeffectormolecules AT evvasilyeva phenotypiccharacterisationofperipheralbloodcytotoxictlymphocytesregulatoryandeffectormolecules AT iikrobinets phenotypiccharacterisationofperipheralbloodcytotoxictlymphocytesregulatoryandeffectormolecules AT aasavchenko phenotypiccharacterisationofperipheralbloodcytotoxictlymphocytesregulatoryandeffectormolecules AT mkserebriakova phenotypiccharacterisationofperipheralbloodcytotoxictlymphocytesregulatoryandeffectormolecules AT atotolianareg phenotypiccharacterisationofperipheralbloodcytotoxictlymphocytesregulatoryandeffectormolecules
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PHENOTYPIC CHARACTERISATION OF PERIPHERAL BLOOD CYTOTOXIC T LYMPHOCYTES: REGULATORY AND EFFECTOR MOLECULES

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