Phenotype and function of human natural killer cells purified by using a clinical-scale immunomagnetic method

Department of Hematology-Oncology, St. Jude Children's Research Hospital, Mail Stop 260, 332 N. Lauderdale Street, Memphis, TN 38105, USA.
Cancer Immunology and Immunotherapy (Impact Factor: 3.94). 05/2005; 54(4):389-94. DOI: 10.1007/s00262-004-0609-6
Source: PubMed


Infection, disease relapse, graft failure, and graft-versus-host disease (GVHD) are significant adverse events associated with allogeneic bone marrow transplantation. Donor natural killer (NK) cells may be an ideal cell type for prevention or treatment of all these adverse events. Therefore, we investigated the phenotype and function of human NK cells purified by using a clinical-scale immunomagnetic method. We found that the NK cell purification procedures did not adversely affect the expression of killer cell immunoglobulin-like receptors, adhesion molecules, intracellular cytokines, perforin, and granzyme B. Purified NK cells had extensive proliferative capacity and potent antitumor activity when assessed using an immunodeficient mouse model. While all mice transplanted with unpurified mononuclear cells developed GVHD, none of the mice transplanted with purified NK cells did. NK cells were highly susceptible to lysis by antithymocyte globulin (ATG), whereas G-CSF had a minimal effect on their natural cytotoxicity. These results support future clinical investigation of the use of purified NK cells for adoptive immunotherapy in the absence of ATG.

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Available from: Marti Holladay, Jul 10, 2014
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    • "However, the clinical responses of these studies have been unsatisfactory, and thus further efforts will be needed. Development of ex vivo expansion and activation of NK cells has been extensively assessed by groups at St. Jude Children's Research Hospital (Leung et al., 2005), the National Institutes of Health (Berg et al., 2009), Singapore (Suck et al., 2011), and the Karolinska Institute (Sutlu et al., 2010). These methods have generally been based on CD56-positive selection with or without CD3-depletion/CD3-suppression followed by IL-2 and/or IL-15 stimulation, and in some case virus-infected feeder cells have been used to obtain highly activated NK cells. "
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    ABSTRACT: Adoptive immunotherapy using natural killer (NK) cells has been a promising treatment for intractable malignancies; however, there remain a number of difficulties with respect to the shortage and limited anticancer potency of the effector cells. We here established a simple feeder-free method to generate purified (>90%) and highly activated NK cells from human peripheral blood-derived mononuclear cells (PBMCs). Among the several parameters, we found that simply 1) CD3-depletion, 2) high dose IL-2, and 3) use of a specific culture medium were sufficient to obtain highly purified, expanded (~200-fold) and activated CD3-/CD56+ NK cells from PBMCs that we named zenithal-NK cells (Z-NK). Almost all Z-NK cells expressed the lymphocyte-activated marker CD69, and showed dramatically high expression of activation receptors (i.e., NKG2D), interferon-γ, perforin and granzyme B. Importantly, only 2 hours of reaction at an effector/target ratio of 1:1 was sufficient to kill almost all K562 cells, and the antitumor activity was also replicated in tumor-bearing mice in vivo. Cytolysis was specific for various tumor cells, but not for normal cells, irrespective of MHC class I expression. These findings strongly indicate that Z-NK cells are purified, expanded, and near-fully activated human NK cells and warrant further investigation in a clinical setting.
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    • "A large number of NK cells can be collected from a healthy donor by a single largevolume leukapheresis process. Purified NK cells can be easily prepared using an NK cell-specific antibody, such as anti- CD56, and immunomagnetic methods (Iyengar et al, 2003; Leung et al, 2005a). The end products usually contain no measurable T cells or B cells, thus avoiding GVHD and Epstein-Barr virus-associated lymphoproliferative disease. "
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    ABSTRACT: Analogous to T cells, Natural Killer (NK) cells may facilitate engraftment, combat infection, and control cancer in bone marrow or haematopoietic stem cell transplantation (HSCT); however, NK cells do not cause graft-versus-host disease. Killer immunoglobulin-like receptors (KIRs) regulate NK cell function, and recent data suggest that KIR is as important as its ligand (human leucocyte antigen; HLA) in HSCT for both malignant and non-malignant conditions. Because there is substantial variability in KIR gene content, allelic polymorphism, and cell-surface expression among people, careful selection of donors based on HLA and KIR is essential to optimize HSCT outcomes. Furthermore, NK cells may be used for adoptive immunotherapy after HSCT in place of conventional donor lymphocyte infusion, as part of pre-transplant cytoreductive therapy, or as an independent therapeutic agent in high-risk leukaemia in place of sibling donor HSCT.
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    • "Therefore, it is logical to examine the application of the receptor – ligand mismatch model to autologous HCTs. Because lymphoid leukaemias and solid tumours such as neuroblastoma are susceptible to KIR – HLA mismatched NK cell – mediated lysis (Leung et al, 2004, 2005a), we conducted a prospective study to test the hypothesis that patients with lymphoma or other solid tumour who receive autologous HCT have a low risk of relapse if KIR – HLA mismatch(es) is present. "
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    ABSTRACT: Genes that encode killer Ig-like receptors (KIRs) and their HLA class I ligands segregate independently; thus, some individuals may express an inhibitory KIR gene but not its cognate ligand. We hypothesised that these patients with KIR-HLA receptor-ligand mismatch have a low risk of relapse after an autologous haematopoietic stem cell transplantation (HCT). Sixteen consecutive patients with lymphoma or solid tumour were enrolled onto a prospective study. They received high-dose busulphan and melphalan followed by autologous CD133(+) HCT. We found that 8 of the 16 patients experienced disease progression after autologous HCT, including 5 of the 6 patients (83%) with no inhibitory KIR-HLA mismatch and 3 of the 6 patients (50%) with 1 mismatched pair; none of the 4 (0%) patients with 2 mismatched pairs experienced disease progression. Survival analyses showed that inhibitory KIR-HLA mismatch was the only significant prognostic factor (P=0.01). The potential applicability of the receptor-ligand mismatch model to autologous HCTs and to patients with lymphoma or solid tumour is clinically significant because of the prevalence of the HCT procedure.
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