Iranian Journal of Immunology

Leukocyte Reduction Filters Are Reliable and Economic Source for Natural Killer Cell Preparation

Document Type : Original Article

Authors

Moosa Rezwani 1
Abdulbaset Mazarzaei 1
Zahra Abbasi-Malati 2
Ali Akbar Pourfathollah 1

1 Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

2 Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.

https://doi.org/10.22034/iji.2023.92527.2158
Abstract
Background: An issue that hinders researchers’ access to Natural Killer (NK) cells is their low proportion in peripheral blood leukocytes. This issue is currently addressed by methods involving a series of differentiation and expansions that are time-consuming and expensive.
Objective: We have investigated whether the used leukocyte reduction filters, a by-product in the blood transfusion practice that currently is considered waste, can be utilized as a source of the NK cells.
Methods: Following the blood donation of 46 donors based on the Iranian Blood Transfusion Organization’s protocols, a sample of peripheral blood of each donor and the leukocyte reduction filter used in their donation procedure have been obtained. The entrapped cells were flushed back from the leukocyte reduction filters. Both groups of samples were analyzed using an automatic hematological analyzer. NK cell isolation was done by the MACS negative selection method. The samples have been comparatively analyzed utilizing flow cytometry data of NK cells’ subpopulation compositions, viability, degranulation patterns, and cytotoxic capacity against the K562 cell line.
Results: Every major leukocyte population was abundant in the samples extracted from the used leukocyte reduction filters. The NK cells extracted from leukocyte reduction filters did not show any statistically meaningful differences (P<0.5) from peripheral blood samples in terms of subpopulation composition, viability, degranulation potency, and cytotoxic capacity.
Conclusion: Used leukocyte reduction filters can be considered an economic, easy to obtain, and robust source of abundant research-grade NK cells.

Keywords

Cell Isolation
Leukocyte Reduction Procedures
Natural Killer Cells
Peripheral Blood Mononuclear Cells
  1. Fujisaki H, Kakuda H, Shimasaki N, Imai C, Ma J, Lockey T, et al. Expansion of highly cytotoxic human natural killer cells for cancer cell therapy. Cancer Res. 2009;69(9):4010–7.
  2. Lapteva N, Durett AG, Sun J, Rollins LA, Huye LL, Fang J, et al. Large-scale ex vivo expansion and characterization of natural killer cells for clinical applications. Cytotherapy. 2012 Oct 1;14(9):1131–43.
  3. Gong JH, Maki G, Klingemann HG. Characterization of a human cell line (NK-92) with phenotypical and functional characteristics of activated natural killer cells. Leukemia. 1994 Apr;8(4):652–8.
  4. Spanholtz J, Preijers F, Tordoir M, Trilsbeek C, Paardekooper J, Witte T de, et al. Clinical-Grade Generation of Active NK Cells from Cord Blood Hematopoietic Progenitor Cells for Immunotherapy Using a Closed-System Culture Process. PLOS ONE. 2011 Jun 16;6(6):e20740.
  5. Research C for BE and. Pre-Storage Leukocyte Reduction of Whole Blood and Blood Components Intended for Transfusion [Internet]. U.S. Food and Drug Administration. FDA; 2019 [cited 2020 Nov 26]. Available from: https://www.fda.gov/regulatory-information/search-fda-guidance-documents/pre-storage-leukocyte-reduction-whole-blood-and-blood-components-intended-transfusion
  6. Sharma RR, Marwaha N. Leukoreduced blood components: Advantages and strategies for its implementation in developing countries. Asian J Transfus Sci. 2010;4(1):3–8.
  7. Dezfouli AB, Pourfathollah AA, Nikougoftar-Zarif M, Khosravi M, Tajrishi M, Ezzati N, et al. Optimizing the recovery of peripheral blood mononuclear cells trapped in leukoreduction filters - A comparison study. Hematol Transfus Cell Ther [Internet]. 2020 Dec 21 [cited 2020 Dec 30]; Available from: http://www.sciencedirect.com/science/article/pii/S253113792031302X
  8. Meyer TPH, Zehnter I, Hofmann B, Zaisserer J, Burkhart J, Rapp S, et al. Filter Buffy Coats (FBC): a source of peripheral blood leukocytes recovered from leukocyte depletion filters. J Immunol Methods. 2005;307(1–2):150–66.
  9. Valiathan R, Deeb K, Diamante M, Ashman M, Sachdeva N, Asthana D. Reference ranges of lymphocyte subsets in healthy adults and adolescents with special mention of T cell maturation subsets in adults of South Florida. Immunobiology. 2014 Jul 1;219(7):487–96.
  10. Bisset LR, Lung TL, Kaelin M, Ludwig E, Dubs RW. Reference values for peripheral blood lymphocyte phenotypes applicable to the healthy adult population in Switzerland. Eur J Haematol. 2004 Mar 1;72(3):203–12.
  11. Chng WJ, Tan GB, Kuperan P. Establishment of Adult Peripheral Blood Lymphocyte Subset Reference Range for an Asian Population by Single-Platform Flow Cytometry: Influence of Age, Sex, and Race and Comparison with Other Published Studies. Clin Diagn Lab Immunol. 2004 Jan 1;11(1):168–73.
  12. Shah N, Martin-Antonio B, Yang H, Ku S, Lee DA, Cooper LJN, et al. Antigen Presenting Cell-Mediated Expansion of Human Umbilical Cord Blood Yields Log-Scale Expansion of Natural Killer Cells with Anti-Myeloma Activity. PLOS ONE. 2013 Oct 18;8(10):e76781.
  13. Yoon SR, Lee YS, Yang SH, Ahn KH, Lee J-H, Lee J-H, et al. Generation of donor natural killer cells from CD34 + progenitor cells and subsequent infusion after HLA-mismatched allogeneic hematopoietic cell transplantation: a feasibility study. Bone Marrow Transplant. 2010 Jun;45(6):1038–46.
  14. Luevano M, Madrigal A, Saudemont A. Generation of natural killer cells from hematopoietic stem cells in vitro for immunotherapy. Cell Mol Immunol. 2012 Jul;9(4):310–20.
  15. Eguizabal C, Zenarruzabeitia O, Monge J, Santos S, Vesga MA, Maruri N, et al. Natural Killer Cells for Cancer Immunotherapy: Pluripotent Stem Cells-Derived NK Cells as an Immunotherapeutic Perspective. Front Immunol [Internet]. 2014 [cited 2020 Nov 20];5. Available from: https://www.frontiersin.org/articles/10.3389/fimmu.2014.00439/full
  16. Woll PS, Grzywacz B, Tian X, Marcus RK, Knorr DA, Verneris MR, et al. Human embryonic stem cells differentiate into a homogeneous population of natural killer cells with potent in vivo antitumor activity. Blood. 2009 Jun 11;113(24):6094–101.
  17. Yodoi J, Teshigawara K, Nikaido T, Fukui K, Noma T, Honjo T, et al. TCGF (IL 2)-receptor inducing factor(s). I. Regulation of IL 2 receptor on a natural killer-like cell line (YT cells). J Immunol. 1985 Mar 1;134(3):1623–30.
  18. Yagita M, Huang CL, Umehara H, Matsuo Y, Tabata R, Miyake M, et al. A novel natural killer cell line (KHYG-1) from a patient with aggressive natural killer cell leukemia carrying a p53 point mutation. Leukemia. 2000 May;14(5):922–30.
  19. Robertson MJ, Cochran KJ, Cameron C, Le JM, Tantravahi R, Ritz J. Characterization of a cell line, NKL, derived from an aggressive human natural killer cell leukemia. Exp Hematol. 1996;24(3):406–15.
  20. Klingemann H, Boissel L, Toneguzzo F. Natural Killer Cells for Immunotherapy – Advantages of the NK-92 Cell Line over Blood NK Cells. Front Immunol [Internet]. 2016 [cited 2020 Nov 18];7. Available from: https://www.frontiersin.org/articles/10.3389/fimmu.2016.00091/full#B13
  21. Maki G, Klingemann H-G, Martinson JA, Tam YK. Factors Regulating the Cytotoxic Activity of the Human Natural Killer Cell Line, NK-92. J Hematother Stem Cell Res. 2001 Jun 1;10(3):369–83.
  22. Fujisaki H, Kakuda H, Imai C, Mullighan CG, Campana D. Replicative potential of human natural killer cells. Br J Haematol. 2009;145(5):606–13.
  23. World Health Organization. WHO: Blood safety and availability [Internet]. [cited 2020 Nov 24]. Available from: https://www.who.int/news-room/fact-sheets/detail/blood-safety-and-availability
  24. Longley RE, Stewart D. Recovery of functional human lymphocytes from Leukotrap filters. J Immunol Methods. 1989 Jul 6;121(1):33–8.
  25. Weitkamp J-H, Crowe Jr JE. Blood donor leukocyte reduction filters as a source of human B lymphocytes. Biotechniques. 2001;31(3):464–6.
  26. Izquierdo N, Naranjo M, Fernández M, Cos J, Massuet L, Martínez-Picado J, et al. Leukocyte reduction filters: An alternative source of peripheral blood mononuclear cells. Inmunología. 2003;22(3):255–62.
  27. Tremblay MM, Houtman JCD. TCR-mediated functions are enhanced in activated peripheral blood T cells isolated from leucocyte reduction systems. J Immunol Methods. 2015 Jan 1;416:137–45.
  28. Ebner S, Neyer S, Hofer S, Nussbaumer W, Romani N, Heufler C. Generation of large numbers of human dendritic cells from whole blood passaged through leukocyte removal filters: an alternative to standard buffy coats. J Immunol Methods. 2001 Jun 1;252(1):93–104.
  29. Valizadeh M, Purfathollah AA, Raoofian R, Homayoonfar A, Moazzeni M. Optimized simple and affordable procedure for differentiation of monocyte-derived dendritic cells from LRF: An accessible and valid alternative biological source. Exp Cell Res. 2021 Sep 15;406(2):112754.
  30. Ivanovic Z, Duchez P, Morgan DA, Hermitte F, Lafarge X, Chevaleyre J, et al. Whole‐blood leukodepletion filters as a source of CD34+ progenitors potentially usable in cell therapy. Transfusion (Paris). 2006;46(1):118–25.
  31. Peytour Y, Guitart A, Villacreces A, Chevaleyre J, Lacombe F, Ivanovic Z, et al. Obtaining of CD34+ cells from healthy blood donors: development of a rapid and efficient procedure using leukoreduction filters. Transfusion (Paris). 2010;50(10):2152–7.
  32. Peytour Y, Villacreces A, Chevaleyre J, Ivanovic Z, Praloran V. Discarded leukoreduction filters: a new source of stem cells for research, cell engineering and therapy? Stem Cell Res. 2013;11(2):736–42.
  33. Ferdowsi S, Pourfathollah AA, Amiri F, Rafiee MH, Aghaei A. Evaluation of anticancer activity of α-defensins purified from neutrophils trapped in leukoreduction filters. Life Sci. 2019 May 1;224:249–54.
  34. Teleron AA, Carlson B, Young PP. Blood donor white blood cell reduction filters as a source of human peripheral blood–derived endothelial progenitor cells. Transfusion (Paris). 2005;45(1):21–5.
  35. Ferdowsi S, Abbasi-Malati Z, Pourfathollah AA. Leukocyte reduction filters as an alternative source of peripheral blood leukocytes for research. Hematol Transfus Cell Ther [Internet]. 2020 Dec 24 [cited 2020 Dec 30]; Available from: http://www.sciencedirect.com/science/article/pii/S2531137920313043
  36. Vossier L, Leon F, Bachelier C, Marchandin H, Lehmann S, Leonetti J-P, et al. An innovative biologic recycling process of leukoreduction filters to produce active human antimicrobial peptides. Transfusion (Paris). 2014;54(5):1332–9.
  37. Sasani N, Roghanian R, Emtiazi G, Aghaie A. A Novel Approach on Leukodepletion Filters: Investigation of Synergistic Anticancer Effect of Purified α-Defensins and Nisin. Vol. 11, Adv Pharm Bull. 2021. p. 378–84.
  38. Cook MA, Jobson SE, Atkinson DC, Lowe DP, Farmer SL, Alvi‐Ali WJ, et al. Used leucodepletion filters as a source of large quantities of DNA suitable for the study of genetic variations in human populations. Transfus Med. 2003;13(2):77–82.
  39. Néron S, Dussault N, Racine C. Whole‐blood leukoreduction filters are a source for cryopreserved cells for phenotypic and functional investigations on peripheral blood lymphocytes. Transfusion (Paris). 2006;46(4):537–44.
Iranian Journal of Immunology
Volume 20, Issue 1
Winter 2023
Pages 114-128

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