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...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | Russian |
| Published: |
St. Petersburg branch of the Russian Association of Allergologists and Clinical Immunologists
2018-03-01
|
| Series: | Медицинская иммунология |
| Subjects: | |
| Online Access: | https://www.mimmun.ru/mimmun/article/view/1485 |
| _version_ | 1797197973740847104 |
|---|---|
| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-08T05:49:01Z |
| format | Article |
| id | doaj.art-bcfc3ee5581440e3a5839c244882d677 |
| institution | Directory Open Access Journal |
| issn | 1563-0625 2313-741X |
| language | Russian |
| last_indexed | 2024-04-24T06:52:29Z |
| publishDate | 2018-03-01 |
| publisher | St. Petersburg branch of the Russian Association of Allergologists and Clinical Immunologists |
| record_format | Article |
| series | Медицинская иммунология |
| spelling | doaj.art-bcfc3ee5581440e3a5839c244882d6772024-04-22T13:07:42ZrusSt. Petersburg branch of the Russian Association of Allergologists and Clinical ImmunologistsМедицинская иммунология1563-06252313-741X2018-03-0120222724010.15789/1563-0625-2018-2-227-2401001PHENOTYPIC CHARACTERISATION OF PERIPHERAL BLOOD CYTOTOXIC T LYMPHOCYTES: REGULATORY AND EFFECTOR MOLECULESI. V. Kudryavtsev0A. G. Borisov1E. V. Vasilyeva2I. I. Krobinets3A. A. Savchenko4M. K. Serebriakova5A. Totolian Areg6Institute of Experimental Medicine; First St. Petersburg State I. Pavlov Medical UniversityResearch Institute of Medical Problems of the North, Krasnoyarsk Science Center, Siberian Branch of Russian Academy of Sciences; Krasnoyarsk State V.F. Voino-Yasenetsky Medical UniversitySt. Petersburg Pasteur InstituteRussian Research Institute of Hematology and TransfusiologyResearch Institute of Medical Problems of the North, Krasnoyarsk Science Center, Siberian Branch of Russian Academy of Sciences; Krasnoyarsk State V.F. Voino-Yasenetsky Medical UniversityInstitute of Experimental MedicineFirst St. Petersburg State I. Pavlov Medical University; St. Petersburg Pasteur InstituteCytotoxic 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.https://www.mimmun.ru/mimmun/article/view/1485flow cytometrymulticolor immunophenotypingcytotoxic t cell subsetscd3+cd8+ maturationeffector moleculesinhibitory receptors |
| spellingShingle | 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 Медицинская иммунология flow cytometry multicolor immunophenotyping cytotoxic t cell subsets cd3+cd8+ maturation effector molecules inhibitory receptors |
| title | 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_short | 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 |
| topic | flow cytometry multicolor immunophenotyping cytotoxic t cell subsets cd3+cd8+ maturation effector molecules inhibitory receptors |
| url | https://www.mimmun.ru/mimmun/article/view/1485 |
| 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 |