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Protein Fractions of Natural Killer Cell Lysates Affect the Phenotype, Proliferation and Migration of Endothelial Cells in vitro

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Abstract

Each stage of angiogenesis depends on the nature of the communication between endothelial cells and the microenvironment, and on the complex balance between stimulators and inhibitors of this process. Natural killer cells secrete a variety of cytokines and chemokines that can influence the microenvironment. The aim of this study was to assess the effect of protein fractions of an NK-92 natural killer cell lysate on the phenotype, proliferation and migration of the EA.hy926 endothelial cell line in an in vitro experimental model. We have shown that CD54 (ICAM-1) receptor expression by EA.hy926 cells after co-cultivation with 10–50 kDa and 30–80 kDa fractions was elevated in comparison with non-activated cells. The relative number of EA.hy926 cells with the CD54+ phenotype also increased after co-cultivation with the above fractions. The other remaining fractions (<20 kDa) had no effect on the expression of CD54, CD34, CD119 (IFNγR1) and CD31 (PECAM-1) receptors and the relative number of EA.hy926 cells. Proliferation and migration of EA.hy926 cells after co-cultivation with the fractions altered in opposite directions and concentration-dependently or remained unchanged. The residual area not occupied by migrated EA.hy926 cells after co-cultivation with the fractions did not always correlate with the intensity of migration and was not inversely proportional to the number of the migrated cells. Light fractions (<10 kDa) had no effect on the proliferation and migration of EA.hy926 cells. Using the MALDI-TOF mass spectrometric analysis, more than 1200 entries of proteins with a variety of functions were identified in the NK-92 natural killer cell lysate. Among those linked to natural killer cell functions are proteins related to signal transduction, cell skeleton, cell metabolism, cell proliferation/adhesion, immune response, and enzymes. Chemokines, cytokines, growth factors, semaphorins, defensins, collectin, ficolin, galectin-3, interferon β, TGFβ, VEGF, TNF ligands, and regulators of apoptosis could be specific agents responsible for the effects of the protein fractions seen on the target cells. These findings may indicate an alternative involvement of natural killer cell effector proteins in their communication with endothelial cells. The model used may also reflect the effect of the intracellular content of natural killer cells on the endothelium in case of their death by necrosis.

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Funding

This study was funded by the Russian Foundation for Basic Research (grant no. 19-015-00218: cell culture studies, preparative liquid chromatography) and the Ministry of Science and Higher Education of the Russian Federation under contract with D.O. Ott Institute of Obstetrics, Gynecology and Reproductive Medicine (R and D State Registration no. 1021062512052-5-3.2.2: spectrophotometry, electrophoresis).

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Authors and Affiliations

  1. Department of Immunology and Cell Interactions, D.O. Ott Institute of Obstetrics, Gynecology, and Reproductive Medicine, St. Petersburg, Russia

    A. V. Korenevsky, T. N. Gert, M. E. Berezkina, S. A. Sinyavin, V. A. Mikhailova, K. L. Markova, S. A. Selkov & D. I. Sokolov

  2. Research and Development Department, State Research Institute of Highly Pure Biopreparations, St. Petersburg, Russia

    A. S. Simbirtsev

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  1. A. V. Korenevsky
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  2. T. N. Gert
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  3. M. E. Berezkina
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  4. S. A. Sinyavin
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  5. V. A. Mikhailova
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  6. K. L. Markova
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  7. A. S. Simbirtsev
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  8. S. A. Selkov
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  9. D. I. Sokolov
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Contributions

A.K., D.S.: study conception and design; A.S., S.S.: providing key and unique reagents; T.G., M.B., S.S., V.M., and K.M.: conducting experiments, experimental data analysis, and statistical data processing; A.K.: writing and editing a manuscript; S.S.: study supervision; D.S., S.S.: revising and approving a manuscript.

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Correspondence to A. V. Korenevsky.

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Translated by A. Polyanovsky

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The online version contains supplementary material available at https://doi.org/10.1134/S0022093022070171.

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Korenevsky, A.V., Gert, T.N., Berezkina, M.E. et al. Protein Fractions of Natural Killer Cell Lysates Affect the Phenotype, Proliferation and Migration of Endothelial Cells in vitro. J Evol Biochem Phys 58 (Suppl 1), S134–S150 (2022). https://doi.org/10.1134/S0022093022070171

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