Regulatory cytokine effects in vitro on the phenotype of subpopulations CD62L+CD63-, CD62L+CD63+ and microbicidal activity of neutrophilic granulocytes in patients with colorectal cancer

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Relevance. Neutrophilic granulocytes (NG) are the first cells of the immune system to migrate to the tumor and are actively involved in the implementation of a full-fledged antitumor response through the mechanisms of direct killing of tumor cells, extracellular lysis (NET), and through the activation of antibody-dependent cellular cytotoxicity (ADCC), inhibition of angiogenesis, initiation of other cells with antitumor activity. The aim of the study was to study the effect of cytokines IFNα, IFNγ, G-CSF on the CD62L+CD63- and CD62L+CD63+ subsets and the microbicidal activity of NGs in patients with colorectal cancer (CRC) in vitro. Materials and methods. We studied samples of peripheral blood (PB) of 10 patients of both sexes 38-70 years old with newly diagnosed untreated CRC stage II-III (study group) and 10 healthy volunteers (comparison group). The subsets CD62L+CD63+ NG, CD62L+CD63- NG were assessed by flow cytometry (CYTOMICS FC500, Beckman Coulter, USA), the microbicidal functions of NG were tested by cytochemical methods: activity of NADPH - oxidases, myeloperoxidase (MP), level of cationic protein (CP) in spontaneous tests and under additional stress of S. aureus . The effect of IFNα, IFNγ, G-CSF cytokines on subsets and the microbicidal activity of NG in vitro was studied in both study groups. Microsoft Exel 2016 and StatPlus 2010 were used for statistical processing of the obtained data using nonparametric tests: Me (Q1; Q3), Mann-Whitney U-test and Wilcoxon test . Results . The features of transformation of CD62L+ CD63-NG and CD62L+ CD63+ NG subsets of PB in CRC have been established, that allows to get an idea of the NG ability to roll and readiness to activate the microbicidal arsenal, various defects of spontaneous and induced microbicidal activity of oxygendependent and oxygen-independent mechanisms of NG. The effects of cytokine influence on NG in CRC in vitro have been shown, which indicates the possibility of regulating the receptor and microbicidal functions of NG, and, on the other hand, suggests defects in NG perception of regulatory stimuli, that is confirmed by the progression of tumor growth.

About the authors

G. A. Chudilova

Kuban State Medical University

Author for correspondence.
SPIN-code: 2092-6412
Krasnodar, Russian Federation

I. V. Nesterova

Kuban State Medical University

SPIN-code: 4714-2488
Krasnodar, Russian Federation

S. V. Kovaleva

Kuban State Medical University

SPIN-code: 8289-5342
Krasnodar, Russian Federation

L. V. Lomtatidze

Kuban State Medical University

SPIN-code: 2060-9316
Krasnodar, Russian Federation


  1. Cancer Research UK. Cancer statistics. available at: www. Access date: 10/08/2020.
  2. Mantovani A., Romero P., Palucka A.K., Marincola F.M. Tumour immunity: effector response to tumor and role of the microenvironment. Lancet. 2008; 371:771–783.
  3. McLean M.H., Murray G.I., Stewart K.N., Norrie G., Mayer C., Hold G.L., Thomson J., Fyfe H., Mowat N.A., Drew E.J., El-Omar M.E. The inflammatory microenvironment in colorectal neoplasia. PLoS One. 2011;6(1): e15366. doi: 10.1371/journal.pone.0015366.
  4. Swierczak A., Mouchemore K.A., Hamilton J.A., Anderson R.L. Neutrophils: important contributors to tumor progression and metastasis. Cancer Metastasis. 2015;34:735–751. doi:10.1007/ s10555–015–9594–9
  5. Sionov R.V., Fridlender Z.G., Granot Z. The multifaceted roles neutrophils play in the tumor microenvironment. Cancer Microenviron. 2015;8:125–158. doi: 10.1007/s12307–014–0147–5
  6. Coffelt S.B., Wellenstein M.D., de Visser K.E. Neutrophils in cancer: neutral no more. Nat. Rev. Cancer. 2016;16:431–446. doi: 10.1038/nrc.2016.52
  7. Mishalian I., Granot Z., Fridlender Z.G. The diversity of circulating neutrophils in cancer. Immunobiology. 2017;222:82–88. doi: 10.1016/j.imbio.2016.02.001
  8. Kabayashi Y. The role of chemokines in neutrophil biology Frontiers in bioscience: a journal and virtual library. 2008;13:2400–2407.
  9. Khajah M., Millen B., Cara D.C., Waterhouse C., Mc Cafferty D. Granulocyte-macrophage colony-stimulating factor (GM–CSF): a chemoattractive agent for murine leukocytes in vivo. Journal of Leukocyte Biology. 2011; 89 (6):945–953.
  10. Dumitru C.A., Lang S., Brandau S. Modulation of neutrophil granulocytes in the tumor microenvironment: Mechanisms and consequences for tumor progression. Seminars in Cancer Biology. 2013;23:141–148.
  11. Mishalian I., Granot Z., Fridlender Z.G. The diversity of circulating neutrophils in cancer.Immunobiology. 2017;222 (1):82–88.
  12. Zhang X., Zhang W., Yuan X., Fu M., Qian H., Xu W. Neutrophils in cancer development and progression: Roles, mechanisms, and implications (Review). International journal of oncology. 2016; 49(3):857–867.
  13. Lau D., Mollnau H., Eiserich J.P., Freeman B.A., Daiber A., Gehling U.M., Brümmer J., Rudolph V., Münzel T., Heitzer T., Meinertz T., Baldus S. Myeloperoxidase mediates neutrophil activation by association with CD11b/CD18 integrins. Proceedings of the National Academy of Sciences. 2005;102 (2):431–436.
  14. Mantovani А. The yin-yang of tumor-associated neutrophils. Cancer Cell. 2009;8:173–174.
  15. Levy O. Innate immunity of the newborn: basic mechanisms clinical correlates. Nature reviews Immunology. 2007;7(5): 379–390.
  16. Lin A., Lore K. Granulocytes: New Members of the AntigenPresenting Cell Family. Frontiers in Immunology. 2017;11. doi: 10.3389/fimmu.2017.01781
  17. Dallegri F., Ottonello L., Ballesterro A., Patrizia Dapino, F. Ferrando, F. Patrone, C. Sacchetti Tumor cell lysis by activated human neutrophils: analysis of neutrophil-delivered oxidative attack and role of leukocyte function-associated antigen. Inflammation. 1991; 1(15):15–30.
  18. Lai X., Gu Q., Zhou X., Feng W., Lin X., He Y., Cao J., Liu P., Zhang H., Zheng X. Decreased expression of CD63 tetraspanin protein predicts elevated malignant potential in human esophageal cancer. Oncol Lett. 2017;13(6):4245–4251. doi: 10.3892/ol.2017.6023.
  19. Liu W.H., Li X., Zhu X.L., Hou M.L., Zhao W. CD63 inhibits the cell migration and invasion ability of tongue squamous cell carcinoma. Oncol Lett. 2018;(6): 9033–9042. DOI: 10.3892 / ol.2018.8499.
  20. Beinert T., Munzing S., Possinger K., Krombach F. Increased expression of the tetraspanins CD53 and CD63 on apoptotic human neutrophils.Journal of Leukocyte Biology. 2000; 67:369–373.
  21. Droeser R.A., Hirt C., Eppenberger-Castori S., Zlobec I., Viehl C.T., Frey D.M., Nebiker C.A., Rosso R., Zuber M., Amicarella F. High myeloperoxidase positive cell infiltration in colorectal cancer is an independent favorable prognostic factor. PLoS One. 2013; 8: e64814.
  22. Roncucci L., Mora E., Mariani F., Bursi S., Pezzi A., Rossi G., Pedroni M., Luppi D., Santoro L., Monni S. Myeloperoxidasepositive cell infiltration in colorectal carcinogenesis as indicator of colorectal cancer risk. Cancer Epidemiol Biomarkers Prev. 2008; 17: 2291–2297.
  23. Rao H.L., Chen J.W., Li M., Xiao Y.B., Fu J., Zeng Y.X., Cai M.Y., Xie D. Increased intratumoral neutrophil in colorectal carcinomas correlates closely with malignant phenotype and predicts patients’ adverse prognosis. PLoS One. 2012;7: e30806
  24. Gustafson M.P., Lin Y., Maas M.L., Van Keulen V.P., Johnston P.B., Peikert T., Gastineau D.A., Dietz A.B. A method for identification and analysis of non-overlapping myeloid immunophenotypes in humans. PLoS One. 2015; 10: e0121546.



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Copyright (c) 2020 Chudilova G.A., Nesterova I.V., Kovaleva S.V., Lomtatidze L.V.

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