Evaluation of Neutrophils Immunophenotype in the Microenvironment of Malignant Pleural Effusions

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The lung cancer is often associated with the development of pleural effusion. Neutrophils are the most numerous population of immune system cells which are an essential component of tumor leukocyte infiltration. These cells are engaged in the development and maintenance of the inflammation. It is indicated that neutrophils support the development of cancer. The aim of the study was the evaluation of neutrophils, regarding their presence and activity in pleural effusions. This was achieved by assessing of molecular structures, which are used by neutrophils in chemotaxis and phagocytosis. 60 pleural effusions and 34 peripheral blood samples received from patients and 15 peripheral blood samples from the control group were analyzed. Expression of CD11a, CD11b, CD11c, CD18 and CD62L molecules with use flow cytometry was evaluated. The concentration of the neutrophil elastase in pleural effusions were measured with use ELISA test. The number of neutrophils in the peripheral blood of patients with pleural effusion was lower than that observed in the control group. Neutrophils present in pleural effusions were characterized by an increased ability to chemotaxis and secrete significant amounts of neutrophil elastase. Neutrophils recruited into the pleura during the formation of the effusion are an essential element of the developing inflammatory reaction in this environment. The presence of neutrophils in pleural effusion may promote its further formation and support the development of cancer.

1. Karkhanis VS, Joshi JM. Pleural effusion: diagnosis, treatment, and management. Open Access Emerg Med. 2012;4:31-52; DOI:10.2147/OAEM. S29942.

2. Ried M, Hofmann HS. The treatment of pleural carcinosis with malignant pleural effusion. Dtsch Arztebl Int. 2013;110(18):313-18; DOI:10.3238/arztebl.2013.0313.

3. Noppen M, De Waele M, Li R, Gucht KV, D’Haese J, Gerlo E, Vincken W. Volume and cellular content of normal pleural fluid in humans examined by pleural lavage. Am J Respir Crit Care Med. 2000;162(3Pt1):1023-6; DOI:10.1164/ajrccm.162.3.9910050.

4. Light RW, Erozan YS, Ball CW. Cells in the pleural fluid: their value in differential diagnosis. Arch Intern Med. 1973;132(6):854-60; DOI:10.1001/ archinte.1973.03650120060011.

5. Bruehl RE, Moore KL, Lorant DE, Borregaard N, Zimmerman GA, McEver RP, Bainton DF. Leukocyte activation induces surface redistribution of P-selectin glycoprotein ligand-1. J Leukoc Biol. 1997;61(4):489-99; DOI:10.1002/jlb.61.4.489

6. Abram CL, Lowell CA. The ins and outs of leukocyte integrin signaling. Annu Rev Immunol. 2009;27:339-62; DOI:10.1146/annurev. immunol.021908.132554.

7. Etzioni A. Adhesion Molecules-Their Role in Health and Disease. Pediatr Res. 1996;39(2):191-8; DOI: 10.1203/00006450-199604001-01156.

8. Kolaczkowska E, Kubes P. Neutrophil recruitment and function in health and inflammation. Nat Rev Immunol. 2013;13(3):159-75; DOI:10.1038/ nri3399.

9. Repp R, Valerius T, Sendler A, Gramatzki M, Iro H, Kalden JR, Platzer E. Neutrophils express the high affinity receptor for IgG (Fc RI, CD64) after in vivo application of recombinant human granulocyte colony-stimulating factor. Blood. 1991;78(4):885-9.

10. Lukácsi S, Nagy-Baló Z, Erdei A, Sándor N, Bajtay Z. The role of (CD11b/CD18) and CR4 (CD11c/CD18) in complement-mediated phagocytosis and podosome formation by human phagocytes. Immunol Lett. 2017;189:64-72; DOI:10.1016/j.imlet.2017.05.014.

11. Robinson JM. Reactive oxygen species in phagocytic leukocyte. Histochem Cell Biol. 2008;130(2):281-97; DOI:10.1007/s00418-008-0461-4.

12. Weinrauch Y, Drujan D, Shapiro SD , Weiss J, Zychlinsky A. Neutrophil elastase targets virulence factors of enterobacteria. Nature. 2002;417(6884):91-4; DOI:10.1038/417091a.

13. Janoff A, Schere J. Mediators of inflammation in leukocyte lysosomes. IX. Elastinolytic activity in granules of human polymorphonuclear leukocytes. J Exp Med. 1968;128(5):1137-55; DOI:10.1084/jem.128.5.1137.

14. McDonald JA, Kelly DG. Degradation of fibronectin by human leukocyte elastase: release of biologically active fragments. J Biol Chem. 1980;255(18):8848-58.

15. Mainardi C, Dixit S, Kang A. Degradation of type IV collagen (basement membrane) by a proteinase isolated from human polymorphonuclear leukocyte granules. J Biol Chem. 1980;255(11):5435-41.

16. Papayannopoulos V, Metzler KD, Hakkim A, Zychlinsky A. Neutrophil elastase and myeloperoxidase regulate the formation of neutrophil extracellular traps. J Cell Biol. 2010;191(3):677-91; DOI:10.1083/ jcb.201006052.

17. Sarraf KM, Belcher E, Raevsky E, Nicholson AG, Goldstraw P, Lim E. Neutrophil/ lymphocyte ratio and its association with survival after complete resection in non-small cell lung cancer. J Thorac Cardiovasc Surg. 2009;137(2):425-8; DOI:10.1016/j.jtcvs.2008.05.046.

18. Wu G, Yao Y, Bai C, Zeng J, Shi D, Gu X, Shi X, Song Y. Combination of platelet to lymphocyte ratio and neutrophil to lymphocyte ratio is a useful prognostic factor in advanced non-small cell lung cancer patients. Thorac Cancer. 2015;6(3):275-87; DOI:10.1111/1759-7714.12178.

19. Halazun KJ, Hardy MA, Rana AA, Woodland DC, Luyten EJ, Mahadev S, Witkowski P, Siegel AB, Brown RS Jr, Emond JC. Negative impact of neu trophil-lymphocyte ratio on outcome after liver transplantation for hepatocellular carcinoma. Ann Surg. 2009;250(1):141-51; DOI:10.1097/ SLA.0b013e3181a77e59.

20. He JR, Shen GP, Ren ZF, Qin H, Cui C, Zhang Y, Zeng YX, Jia WH. Pretreatment levels of peripheral neutrophils and lymphocytes as independent prognostic factors in patients with nasopharyngeal carcinoma. Head Neck. 2012;34(12):1769-76; DOI:10.1002/hed.22008.

21. Schmidt H, Bastjolt L, Geersten P, Christensen IJ, Larsen S, Gehl J, von der Maase H. Elevated neutrophils and monocyte counts in peripheral blood are associated with poor survival in patients with metastatic melanoma: a prognostic model. Br J Cancer. 2005;93(3):273-8; DOI:10.1038/ sj.bjc.6602702

22. Teramukai S, Kitano T, Kishida Y, Kawahara M, Kubota K, Komuta K, Minato K, Mio T, Fujita Y, Yonei T, Nakano K, Tsuboi M, Shibata K, Furuse K, Fukushima M. Pretreatment neutrophil count as an independent prognostic factor in advanced non-small-cell lung cancer: an analysis of Japan Multinational Trial Organisation LC00-03. Eur J Cancer. 2009;45(11):1950-8; DOI:10.1016/j.ejca.2009.01.023.

23. Trellakis S, Bruderek K, Dumitru CA, Gholaman H, Gu X, Bankfalvi A, Scherag A, Hütte J, Dominas N, Lehnerdt GF, Hoffmann TK, Lang S, Brandau S. Polymorphonuclear granulocytes in human head and neck cancer: enhanced inflammatory activity, modulation by cancer cells and expansion in advanced disease. Int J Cancer. 2011;129(9):2183-93; DOI:10.1002/ijc.25892.

24. McCourt M, Wang JH, Sookhai S, Redmond HP. Proinflammatory mediators stimulate neutrophil-directed angiogenesis. Arch Surg. 1999;134(12):1325-31; DOI:10.1001/archsurg.134.12.1325.

25. Coussens LM, Tinkle CL, Hanahan D, Werb Z. MMP-9 supplied by bone marrow-derived cells contributes to skin carcinogenesis. Cell. 2000;103(3):481-90; DOI:10.1016/S0092-8674(00)00139-2.

26. Kaczmarek M, Sikora J. Macrophages in malignant pleural effusions - alternatively activated tumor associated macrophages. Contemp Oncol (Pozn). 2012;16(4):279-84; DOI:10.5114/wo.2012.30054.

27. Spicer JD, McDonald B, Cools-Lartigue JJ, Chow SC, Giannias B, Kubes P, Ferri LE. Neutrophils Promote Liver Metastasis via Mac-1-Mediated Interactions with Circulating Tumor Cells. Cancer Res. 2012;72(16):3919-

27; DOI:10.1158/0008-5472.CAN-11-2393.

28. Ostrand-Rosenberg S, Sinha P. Myeloid-derived suppressor cells: linking inflammation and cancer. J Immunol. 2009;182(8):4499-506; DOI:10.4049/jimmunol.0802740.

29. Hurt B, Schulick R, Edil B, El Kasmi KC, Barnett C Jr. Cancer-promoting mechanisms of tumor-associated neutrophils. Am J Surg. 2017;214(5):938-944; DOI:10.1016/j.amjsurg.2017.08.003.

30. Houghton AM. The paradox of tumor associated neutrophils. Fueling growth tumor with cytotoxic substances. Cell Cycle. 2010;9(9):1732-7; DOI:10.4161/cc.9.9.11297

31. Ho AS, Chen CH, Cheng CC, Wang CC, Lin HC, Luo TY, Lien GS, Chang J. Neutrophil elastase as a diagnostic marker and therapeutic target in colorectal cancers. Oncotarget. 2014;5(2):473-80; DOI:10.18632/ oncotarget.1631.

32. Yamashita J, Ogawa M, Abe M, Hayashi N, Kurusu Y, Kawahara K, Shirakusa T. Tumor neutrophil elastase is closely associated with the direct extension of non-small cell lung cancer into the aorta. Chest. 1997;111(4):885-90; DOI:10.1378/chest.111.4.885.

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