Human NK cells internalize recombinant major stress protein HSP70
Heat shock proteins 70 kDa (HSP70) protect intracellular proteins from the damaging effects of stress factors of various natures. Moreover, HSP70 play an important role in the vital activity of cells under normal physiological conditions, performing chaperone functions. These functions are realized...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | Russian |
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St. Petersburg branch of the Russian Association of Allergologists and Clinical Immunologists
2023-06-01
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| Series: | Медицинская иммунология |
| Subjects: | |
| Online Access: | https://www.mimmun.ru/mimmun/article/view/2837 |
| _version_ | 1797197786074054656 |
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| author | M. A. Shevchenko D. G. Garbuz A. I. Davletshin A. A. Boyko M. B. Evgen'ev A. M. Sapozhnikov |
| author_facet | M. A. Shevchenko D. G. Garbuz A. I. Davletshin A. A. Boyko M. B. Evgen'ev A. M. Sapozhnikov |
| author_sort | M. A. Shevchenko |
| collection | DOAJ |
| description | Heat shock proteins 70 kDa (HSP70) protect intracellular proteins from the damaging effects of stress factors of various natures. Moreover, HSP70 play an important role in the vital activity of cells under normal physiological conditions, performing chaperone functions. These functions are realized in the intracellular space; however, in some cases, these proteins are also found on the cell surface and in the extracellular environment. The causes and mechanisms of HSP70 translocation to the cell surface and secretion into the extracellular space have not yet been well understood, but such an unusual localization of HSP70 activates the immune system. The surface HSP70 and their extracellular pool stimulate the cytotoxic activity of NK cells. However, direct experimental evidence for the internalization of HSP70 molecules by NK cells has not yet been demonstrated. This paper presents the results of the interaction of the extracellular HSP70 pool with NK cells from the peripheral blood. The results demonstrated the confirmation of the internalization of exogenous HSP70 molecules by NK cells. To this end, fluorescently labeled recombinant stress-inducible human HSP70 were obtained. The electrophoretic data indicated the absence of protein degradation during the labeling process, the purity and stability of the modified protein. To assess the interaction of HSP70 with NK cells, the fluorescently labeled HSP70 was added to an in vitro culture of NK cells isolated by magnetic separation from the peripheral blood mononuclear fraction and analyzed by confocal microscopy. This analysis indicated that living NK cells internalize extracellular HSP70 with localization both in lysosomes and in phagosomes. Our experiments illustrated for the first time the process of penetration of the extracellular form of HSP70 into these cells. The results suggest that the activation of NK cells under the action of exogenous HSP70 could be associated with the internalization of these protein molecules. |
| first_indexed | 2024-03-08T05:46:17Z |
| format | Article |
| id | doaj.art-0018c8853e7e4e66973163d3028b5883 |
| institution | Directory Open Access Journal |
| issn | 1563-0625 2313-741X |
| language | Russian |
| last_indexed | 2024-04-24T06:49:30Z |
| publishDate | 2023-06-01 |
| publisher | St. Petersburg branch of the Russian Association of Allergologists and Clinical Immunologists |
| record_format | Article |
| series | Медицинская иммунология |
| spelling | doaj.art-0018c8853e7e4e66973163d3028b58832024-04-22T13:07:50ZrusSt. Petersburg branch of the Russian Association of Allergologists and Clinical ImmunologistsМедицинская иммунология1563-06252313-741X2023-06-0125344745210.15789/1563-0625-HNC-28371698Human NK cells internalize recombinant major stress protein HSP70M. A. Shevchenko0D. G. Garbuz1A. I. Davletshin2A. A. Boyko3M. B. Evgen'ev4A. M. Sapozhnikov5Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of SciencesEngelhardt Institute of Molecular Biology, Russian Academy of SciencesEngelhardt Institute of Molecular Biology, Russian Academy of SciencesShemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of SciencesEngelhardt Institute of Molecular Biology, Russian Academy of SciencesShemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of SciencesHeat shock proteins 70 kDa (HSP70) protect intracellular proteins from the damaging effects of stress factors of various natures. Moreover, HSP70 play an important role in the vital activity of cells under normal physiological conditions, performing chaperone functions. These functions are realized in the intracellular space; however, in some cases, these proteins are also found on the cell surface and in the extracellular environment. The causes and mechanisms of HSP70 translocation to the cell surface and secretion into the extracellular space have not yet been well understood, but such an unusual localization of HSP70 activates the immune system. The surface HSP70 and their extracellular pool stimulate the cytotoxic activity of NK cells. However, direct experimental evidence for the internalization of HSP70 molecules by NK cells has not yet been demonstrated. This paper presents the results of the interaction of the extracellular HSP70 pool with NK cells from the peripheral blood. The results demonstrated the confirmation of the internalization of exogenous HSP70 molecules by NK cells. To this end, fluorescently labeled recombinant stress-inducible human HSP70 were obtained. The electrophoretic data indicated the absence of protein degradation during the labeling process, the purity and stability of the modified protein. To assess the interaction of HSP70 with NK cells, the fluorescently labeled HSP70 was added to an in vitro culture of NK cells isolated by magnetic separation from the peripheral blood mononuclear fraction and analyzed by confocal microscopy. This analysis indicated that living NK cells internalize extracellular HSP70 with localization both in lysosomes and in phagosomes. Our experiments illustrated for the first time the process of penetration of the extracellular form of HSP70 into these cells. The results suggest that the activation of NK cells under the action of exogenous HSP70 could be associated with the internalization of these protein molecules.https://www.mimmun.ru/mimmun/article/view/2837hsp70 – heat shock protein 70 kdank cellsinternalizationfluorescent labelingfluorescent imaginglaser confocal microscopy |
| spellingShingle | M. A. Shevchenko D. G. Garbuz A. I. Davletshin A. A. Boyko M. B. Evgen'ev A. M. Sapozhnikov Human NK cells internalize recombinant major stress protein HSP70 Медицинская иммунология hsp70 – heat shock protein 70 kda nk cells internalization fluorescent labeling fluorescent imaging laser confocal microscopy |
| title | Human NK cells internalize recombinant major stress protein HSP70 |
| title_full | Human NK cells internalize recombinant major stress protein HSP70 |
| title_fullStr | Human NK cells internalize recombinant major stress protein HSP70 |
| title_full_unstemmed | Human NK cells internalize recombinant major stress protein HSP70 |
| title_short | Human NK cells internalize recombinant major stress protein HSP70 |
| title_sort | human nk cells internalize recombinant major stress protein hsp70 |
| topic | hsp70 – heat shock protein 70 kda nk cells internalization fluorescent labeling fluorescent imaging laser confocal microscopy |
| url | https://www.mimmun.ru/mimmun/article/view/2837 |
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