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• W2034287952 abstract "The prognosis after hematopoietic cell transplantation (HCT) for the treatment of leukemia or lymphoma in humans is influenced by donor-derived natural killer (NK) cells, which enhance the graft-versus-leukemia (GVL) effect. Such alloreactive killer cells can be generated in vivo after HCT if the donor expresses killer cell immunoglobulin-like receptors (KIRs), such as KIR2DL1, KIR2DL2/3, or KIR3DL1, for which the recipient lacks HLA class I ligands. We studied effector cells from 22 KIR/HLA-ligand mismatched and 14 KIR/HLA-ligand matched, primarily HLA-matched patient-donor pairs after allogeneic HCT. A novel 8-color flow cytometry panel allowed us to characterize effector-cell populations without “broadly reactive” inhibitory receptors such as CD94/NKG2A or LIR1. The numbers of such NKG2A– LIR1– NK cells increased following HCT in patients transplanted by KIR/HLA-ligand mismatched grafts, compared to KIR/HLA-ligand matched grafts, and in patients transplanted from donors of the A/B, compared to A/A, KIR haplotypes. NKG2A–LIR1– NK cells expressing only those inhibitory KIRs for which the patient had no HLA class I ligands could be stimulated by HLA class I-deficient cells to express CD107a. Thus, NKG2A–LIR1– NK cells may be important GVL effector cells following HCT, even in patients transplanted from HLA-matched donors. The prognosis after hematopoietic cell transplantation (HCT) for the treatment of leukemia or lymphoma in humans is influenced by donor-derived natural killer (NK) cells, which enhance the graft-versus-leukemia (GVL) effect. Such alloreactive killer cells can be generated in vivo after HCT if the donor expresses killer cell immunoglobulin-like receptors (KIRs), such as KIR2DL1, KIR2DL2/3, or KIR3DL1, for which the recipient lacks HLA class I ligands. We studied effector cells from 22 KIR/HLA-ligand mismatched and 14 KIR/HLA-ligand matched, primarily HLA-matched patient-donor pairs after allogeneic HCT. A novel 8-color flow cytometry panel allowed us to characterize effector-cell populations without “broadly reactive” inhibitory receptors such as CD94/NKG2A or LIR1. The numbers of such NKG2A– LIR1– NK cells increased following HCT in patients transplanted by KIR/HLA-ligand mismatched grafts, compared to KIR/HLA-ligand matched grafts, and in patients transplanted from donors of the A/B, compared to A/A, KIR haplotypes. NKG2A–LIR1– NK cells expressing only those inhibitory KIRs for which the patient had no HLA class I ligands could be stimulated by HLA class I-deficient cells to express CD107a. Thus, NKG2A–LIR1– NK cells may be important GVL effector cells following HCT, even in patients transplanted from HLA-matched donors. Allogeneic hematopoietic cell transplantation (HCT) is often the only possibility to cure acute myelogenous leukemias, myelodysplastic syndromes, and aggressive leukemias/lymphomas. A key to survival in this situation is the graft-versus-leukemia (GVL) effect, mediated by effectors from the donor that destroy residual leukemia cells in the patient. These include donor-derived natural killer (NK) cells, which have been shown to play an important role in the outcome of allogeneic HCT [1Ruggeri L. Capanni M. Urbani E. et al.Effectiveness of donor natural killer cell alloreactivity in mismatched hematopoietic transplants.Science. 2002; 295: 2097-2100Crossref PubMed Scopus (2746) Google Scholar, 2Velardi A. Role of KIRs and KIR ligands in hematopoietic transplantation.Curr Opin Immunol. 2008; 20: 581-587Crossref PubMed Scopus (66) Google Scholar]. The function of NK cells is regulated by the balance of signals from clonally distributed inhibitory and activating cell surface receptors [3Parham P. MHC class I molecules and KIRs in human history, health and survival.Nat Rev Immunol. 2005; 5: 201-214Crossref PubMed Scopus (961) Google Scholar]. The same receptors can be expressed on resting or activated γδ and αβ T cells [4Mingari M.C. Vitale C. Cambiaggi A. et al.Cytolytic T lymphocytes displaying natural killer (NK)-like activity: expression of NK-related functional receptors for HLA class I molecules (p58 and CD94) and inhibitory effect on the TCR-mediated target cell lysis or lymphokine production.Int Immunol. 1995; 7: 697-703Crossref PubMed Scopus (212) Google Scholar]. The most important ones are the inhibitory NK receptors, which induce tolerance against self. These include the lectin-like receptor CD94/NKG2A, which is specific for the nonclassical class I molecule HLA-E, and is stabilized by leader peptides from most classical HLA-A, -B, and -C allotypes [5Braud V.M. Allan D.S. O'Callaghan C.A. et al.HLA-E binds to natural killer cell receptors CD94/NKG2A, B and C.Nature. 1998; 391: 795-799Crossref PubMed Scopus (1782) Google Scholar]. The Ig-like receptors, encoded in the leukocyte receptor complex of chromosome 19, can be divided into the killer cell immunoglobulin-like receptors (KIRs) and the leukocyte Ig-like receptors [6Trowsdale J. Genetic and functional relationships between MHC and NK receptor genes.Immunity. 2001; 15: 363-374Abstract Full Text Full Text PDF PubMed Scopus (269) Google Scholar]. The leukocyte Ig-like receptors (CD85), previously called immunoglobulin-like transcripts or leukocyte immunoglobulin-like receptors (LIR), are broadly reactive inhibitory receptors against the HLA-A, -B, and -C allotypes. Similar to CD94/NKG2A, they enable NK cells to survey for overall class I expression [7Cosman D. Fanger N. Borges L. et al.A novel immunoglobulin superfamily receptor for cellular and viral MHC class I molecules.Immunity. 1997; 7: 273-282Abstract Full Text Full Text PDF PubMed Scopus (587) Google Scholar]. In contrast, 3 different KIRs can distinguish between groups of HLA-A, -B, and -C allotypes [8Hansasuta P. Dong T. Thananchai H. et al.Recognition of HLA-A3 and HLA-A11 by KIR3DL2 is peptide-specific.Eur J Immunol. 2004; 34: 1673-1679Crossref PubMed Scopus (265) Google Scholar, 9Moretta A. Bottino C. Vitale M. et al.Activating receptors and coreceptors involved in human natural killer cell-mediated cytolysis.Annu Rev Immunol. 2001; 19: 197-223Crossref PubMed Scopus (1500) Google Scholar, 10Valiante N.M. Uhrberg M. Shilling H.G. et al.Functionally and structurally distinct NK cell receptor repertoires in the peripheral blood of two human donors.Immunity. 1997; 7: 739-751Abstract Full Text Full Text PDF PubMed Scopus (649) Google Scholar]. KIR2DL1 recognize HLA-C allotypes with a lysine at position 80 (C2 ligands such as Cw02, 0307, 0310, 04, 05, or 06). KIR2DL2 and KIR2DL3 recognize HLA-C allotypes with an asparagine at position 80 (C1 ligands, such as Cw01, 03, 07, or 08). KIR3DL1 molecules recognize HLA-B-allotypes with a polymorphic sequence motif at position 77-83 (Bw4 epitope), whereas KIR3DL2 interacts with the HLA-A3 and A11 allotypes. Besides inhibition, NK cells require activation by virus-infected or leukemic cells to mediate their protective functions [11Lanier L.L. Up on the tightrope: natural killer cell activation and inhibition.Nat Immunol. 2008; 9: 495-502Crossref PubMed Scopus (1217) Google Scholar, 12Lanier L.L. Evolutionary struggles between NK cells and viruses.Nat Rev Immunol. 2008; 8: 259-268Crossref PubMed Scopus (379) Google Scholar]. Stimulatory receptors on NK cells include lectin-like receptors such as CD94/NKG2C and the KIRDS molecules. The HLA class I specificities and antibody reactivities of activating KIRs overlap with those of the corresponding inhibitory KIRs. Other activating receptors on NK cells are the NKG2D homodimers whose ligands include MHC class-I-related chain A/B (MICA/B) [13Bahram S. MIC genes: from genetics to biology.Adv Immunol. 2000; 76: 1-60Crossref PubMed Google Scholar]. A third class of activating receptors on NK cells, as well as some T cells, are the natural cytotoxic receptors (NCR), NCR1 (NKp46), NCR2 (NKp44), and NCR3 (NKp30) [9Moretta A. Bottino C. Vitale M. et al.Activating receptors and coreceptors involved in human natural killer cell-mediated cytolysis.Annu Rev Immunol. 2001; 19: 197-223Crossref PubMed Scopus (1500) Google Scholar]. Although the ligands for these receptors have not been fully elucidated, they may be important in the recognition of leukemic cells [9Moretta A. Bottino C. Vitale M. et al.Activating receptors and coreceptors involved in human natural killer cell-mediated cytolysis.Annu Rev Immunol. 2001; 19: 197-223Crossref PubMed Scopus (1500) Google Scholar, 11Lanier L.L. Up on the tightrope: natural killer cell activation and inhibition.Nat Immunol. 2008; 9: 495-502Crossref PubMed Scopus (1217) Google Scholar, 12Lanier L.L. Evolutionary struggles between NK cells and viruses.Nat Rev Immunol. 2008; 8: 259-268Crossref PubMed Scopus (379) Google Scholar]. Inhibitory NK receptor signals generally overrule signaling through stimulatory receptors [10Valiante N.M. Uhrberg M. Shilling H.G. et al.Functionally and structurally distinct NK cell receptor repertoires in the peripheral blood of two human donors.Immunity. 1997; 7: 739-751Abstract Full Text Full Text PDF PubMed Scopus (649) Google Scholar, 14Pende D. Cantoni C. Rivera P. et al.Role of NKG2D in tumor cell lysis mediated by human NK cells: cooperation with natural cytotoxicity receptors and capability of recognizing tumors of nonepithelial origin.Eur J Immunol. 2001; 31: 1076-1086Crossref PubMed Scopus (301) Google Scholar]. Several clinical studies demonstrated improved survival post allogeneic HCT, along with a strong GVL effect against myelogenous leukemias when the patient was lacking 1 of the 3 major KIR/HLA-ligands present in the donor, the Bw4 epitope in HLA-B, or the C1 or C2 ligands in HLA-C [1Ruggeri L. Capanni M. Urbani E. et al.Effectiveness of donor natural killer cell alloreactivity in mismatched hematopoietic transplants.Science. 2002; 295: 2097-2100Crossref PubMed Scopus (2746) Google Scholar, 15Giebel S. Locatelli F. Lamparelli T. et al.Survival advantage with KIR ligand incompatibility in hematopoietic stem cell transplantation from unrelated donors.Blood. 2003; 102: 814-819Crossref PubMed Scopus (466) Google Scholar, 16Beelen D.W. Ottinger H.D. Ferencik S. et al.Genotypic inhibitory killer immunoglobulin-like receptor ligand incompatibility enhances the long-term antileukemic effect of unmodified allogeneic hematopoietic stem cell transplantation in patients with myeloid leukemias.Blood. 2005; 105: 2594-2600Crossref PubMed Scopus (153) Google Scholar, 17Leung W. Iyengar R. Turner V. et al.Determinants of antileukemia effects of allogeneic NK cells.J Immunol. 2004; 172: 644-650PubMed Google Scholar, 18Hsu K.C. Keever-Taylor C.A. Wilton A. et al.Improved outcome in HLA-identical sibling hematopoietic stem-cell transplantation for acute myelogenous leukemia predicted by KIR and HLA genotypes.Blood. 2005; 105: 4878-4884Crossref PubMed Scopus (412) Google Scholar, 19Miller J.S. Cooley S. Parham P. et al.Missing KIR ligands are associated with less relapse and increased graft-versus-host disease (GVHD) following unrelated donor allogeneic HCT.Blood. 2007; 109: 5058-5061Crossref PubMed Scopus (243) Google Scholar, 20Fischer J.C. Ottinger H. Ferencik S. et al.Relevance of C1 and C2 epitopes for hemopoietic stem cell transplantation: role for sequential acquisition of HLA-C-specific inhibitory killer Ig-like receptor.J Immunol. 2007; 178: 3918-3923PubMed Google Scholar]. The likely mechanism for this is that NK cells derived from the graft express a “leukemia-reactive” profile of inhibitory receptors. These inhibitory KIR do not find an HLA class I ligand on the recipient's cells, including residual leukemia cells. Because all KIRs are encoded on chromosome 19, they are segregated independently of the HLA alleles on chromosome 6. Thus, transplants from HLA identical family donors may be KIR-mismatched [21Shilling H.G. McQueen K.L. Cheng N.W. Shizuru J.A. Negrin R.S. Parham P. Reconstitution of NK cell receptor repertoire following HLA-matched hematopoietic cell transplantation.Blood. 2003; 101: 3730-3740Crossref PubMed Scopus (227) Google Scholar], whereas only transplants from identical twins are HLA- and KIR-matched. We studied regenerating NK and T cells following allogeneic HCT in adult patients, most of which were HLA-matched. According to the expression of KIRs in the donor's blood, the patients were divided into KIR/HLA-ligand matched and mismatched groups. In an effort to better understand the GVL effect we focused our analysis on the regenerating NK cells without expression of broadly reactive inhibitory NK receptors (CD94/NKG2A and LIR1 [CD85j]) and monitored their regeneration in KIR/HLA-ligand matched as well as mismatched patients post-HCT. In addition, we monitored NK receptor expression on other innate effector cell populations that may be relevant for the GVL effect postallogeneic HCT, such as NKT cells, NKT-like cells, and γδ T cells. The study included 36 consecutive adult patients treated by allogeneic HCT in the Division of Allogeneic Stem Cell Transplantation, Department of Hematology/Oncology, Freiburg University Hospital. The details of their transplant history are given in Tables 1, S1, and S2. This study was approved by the University of Freiburg Hospital Ethics Committee; all clinical investigations were conducted according to the Declaration of Helsinki Ethical Principles. The patients provided informed written consent prior to inclusion in this study. All patients received granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood progenitor cells from HLA A-, B-, and C-locus matched donors (32 patients) or HLA A-, B-locus matched, but C-locus mismatched donors (4 patients). The patients were only considered C-locus mismatched if the donor and recipient were assigned to different HLA-C allele groups (C1 or C2). For the purpose of this study we assigned the patients to the groups “KIR/HLA-ligand match” and “KIR/HLA-ligand mismatch,” as defined shortly.Table 1Characteristics of the KIR-Ligand Mismatched and KIR-Ligand Matched HCT PatientsKIR-Ligand MismatchKIR-Ligand MatchN2214Age62.5 ± 856.5 ± 15Sex (male / female)18 / 46 / 8Diagnosis (n)AML/MDS = 16AML/MDS = 9B-NHL = 6B-NHL = 5HLA class I mismatch in GVHD direction (n) A00 B00 C31KIR haplotype (n) A/B145 A/A74 B/B15Transplantation (n) Related5 (23%)9 (64%) Unrelated17 (77%)5 (36%)AML indicates acute myelogenous leukemia; MDS, myelodysplastic syndrome; NHL, non-Hodgkin lymphoma; GVHD, graft-versus-host disease; KIR, killer cell immunoglobulin-like receptor. Open table in a new tab AML indicates acute myelogenous leukemia; MDS, myelodysplastic syndrome; NHL, non-Hodgkin lymphoma; GVHD, graft-versus-host disease; KIR, killer cell immunoglobulin-like receptor. Blood samples were drawn from donors and recipients before and after transplantation. The blood samples post-HCT were collected on days 14 (n = 36), 28 (n = 35), 56 (n = 35), 84 (n = 33), 112 (n = 30), 140 (n = 29), 168 (n = 31), 252 (n = 16), and 336 (n = 1). All blood samples were drawn within 1 week of the specified day. In the case of missed collections, samples were obtained at interim time points whenever possible. Three relapsed patients were excluded from further analysis when they had to undergo a second transplantation. All blood samples were freshly processed by isolation of peripheral blood mononuclear cells (PBMCs) on Ficoll gradients (LSM 1077, PAA, Pasching, Austria), followed by staining for flow cytometry analysis. Donor DNA samples were extracted using QIAamp DNA Blood Mini kit (Qiagen, Hilden, Germany). KIR genotyping was performed according to published protocols with minor modifications [20Fischer J.C. Ottinger H. Ferencik S. et al.Relevance of C1 and C2 epitopes for hemopoietic stem cell transplantation: role for sequential acquisition of HLA-C-specific inhibitory killer Ig-like receptor.J Immunol. 2007; 178: 3918-3923PubMed Google Scholar, 22Uhrberg M. Valiante N.M. Shum B.P. et al.Human diversity in killer cell inhibitory receptor genes.Immunity. 1997; 7: 753-763Abstract Full Text Full Text PDF PubMed Scopus (949) Google Scholar] to differentiate inhibitory KIRs from activating KIRs (KIR2DL1/S1, KIR2DL2/S2, KIR2DL3/S3, KIR2DL5, KIR3DL1/S1, KIR3DL2) and determine the KIR haplotypes. The following mouse antihuman monoclonal antibodies (mAbs) were used: CD3-FITC (UCHT1), TCRγδ-FITC (IMMU 510), NKG2A-PE (Z199), CD85j-PE (HP-F1), TCR Vα24-FITC (C15), NKG2D-PE (ON72), NKp30-PE (Z25), NKp44-PE (Z231), NKp46-PE (BAB281), CD3-ECD (UCHT1), CD244-PE-Cy5 (C1.7.1), CD56-PE-Cy7 [N901(NKH-1)], CD158a,h-APC (EB6.B) (all from Beckman Coulter, Krefeld, Germany), and TCRαβ-FITC (BMA031) (Serotec, Oxford, UK). The mAb TCR Vβ11-PE-Cy7 (C21), CD158e-ECD (Z27), and CD158b,j-APC-Cy7 (GL183) were purchased as custom-conjugates (Beckman Coulter, Custom Design Service). For surface staining, PBMC were incubated with different mAb panels, as shown in Table S3. Following 15 minutes of incubation, 50 μL of OptiLyse B solution (Beckman Coulter) were added for red cell lysis. After 15 minutes 500 μL of double-distilled water (Braun, Melsungen, Germany) were added and the tubes were incubated for a further 20 minutes. The samples were washed (PBS + 1% human serum), fixed in 250 μL 1% paraformaldehyde (IOTest 3 Fixative Solution; Beckman Coulter), and events were acquired on a CyAn™ ADP Flow Cytometer (Beckman Coulter). Voltage of the CyAn FlowCytometer was first adjusted using appropriate isotype controls for each color. Then, compensation beads for each color (antimouse Ig CompBeads, BD) were run on the instrument. Upon analysis of the surface markers automatic compensation was preformed using the FlowJo (Tree Star, Ashland, OR, USA) program. PBMC were incubated in complete medium (IMDM, supplemented with L-Arginine, L-Asparagine, L-Glutamine, penicillin, streptomycin [all from Gibco, Invitrogen, Karlsruhe, Germany] and heat-inactivated 10% male human serum [PAN-Biotech, Aidenbach, Germany]) for 4 hours at 37°C in the presence of monensin (GolgiStop; BD) the CD107a-PE-Cy5.5 mAb (H4A3) (Beckman Coulter, Custom Design Service) [23Alter G. Malenfant J.M. Altfeld M. CD107a as a functional marker for the identification of natural killer cell activity.J Immunol Methods. 2004; 294: 15-22Crossref PubMed Scopus (1037) Google Scholar, 24Anfossi N. Andre P. Guia S. et al.Human NK cell education by inhibitory receptors for MHC class I.Immunity. 2006; 25: 331-342Abstract Full Text Full Text PDF PubMed Scopus (922) Google Scholar, 25Uhrberg M. The CD107 mobilization assay: viable isolation and immunotherapeutic potential of tumor-cytolytic NK cells.Leukemia. 2005; 19: 707-709Crossref PubMed Scopus (24) Google Scholar] and K562 cells (50.000 K562/150.000 PBMC). Then, the cells were washed (phosphate-buffered saline [PBS] + 1% human serum), stained for surface markers (30 minutes), fixed, and permeabilized by IntraPrep Permeabilization Reagent (Beckman Coulter). For intracellular staining we used the IFN-γ-Pacific Blue mAb (4S.B3) (eBioscience, Frankfurt, Germany). Following 30 minutes of incubation, the cells were washed, resuspended in 250 μL IOTest 3 Fixative solution (Beckman Coulter) and analyzed (8 colors) on an LSR II cytometer using automatic compensation and the Diva software (BD Biosciences). Donor-recipient pairs were divided into the categories “KIR/HLA-ligand match” and “KIR/HLA-ligand mismatch” according to the receptor (KIR)-ligand (HLA) model [17Leung W. Iyengar R. Turner V. et al.Determinants of antileukemia effects of allogeneic NK cells.J Immunol. 2004; 172: 644-650PubMed Google Scholar, 18Hsu K.C. Keever-Taylor C.A. Wilton A. et al.Improved outcome in HLA-identical sibling hematopoietic stem-cell transplantation for acute myelogenous leukemia predicted by KIR and HLA genotypes.Blood. 2005; 105: 4878-4884Crossref PubMed Scopus (412) Google Scholar]. Using high-resolution HLA typing all patients were assigned to C1, C2, or C1/C2 groups (HLA-C) and to the Bw4 group (HLA-B). In addition, donors' PBMC were examined by multicolor flow cytometry for NK cells expressing KIR2DL1/S1 (C2-specific), KIR2DL2/DL3/S2 (C1-specific), or KIR3DL1/S1 (Bw4-specific) (Table S4). A patient was assigned to the group “KIR/HLA-ligand match” if the KIR expressed by the donor's NK cells matched the patient's HLA-C or -Bw4 ligands. Thus, most patients in the KIR-ligand matched group expressed all 3 KIR ligands C1, C2, and Bw4. In contrast, a patient was assigned to the group “KIR/HLA-ligand mismatch” if the donor had NK cells expressing KIR for which the patient had no HLA-C or Bw4 ligand. This would mean that there was a KIR-mismatch in the graft-versus-host (GVH) direction. Although patients #14 and #18 were missing HLA C2 KIR ligands, they were assigned to the KIR/HLA-ligand match group because phenotypic analysis of their donors' effector cells failed to demonstrate KIR2DL1 expression [26Leung W. Iyengar R. Triplett B. et al.Comparison of killer Ig-like receptor genotyping and phenotyping for selection of allogeneic blood stem cell donors.J Immunol. 2005; 174: 6540-6545PubMed Google Scholar]. For a statistical comparison of the study groups 20,000 viable lymphocytes were collected per sample. All data were analyzed using SPSS software (Version 14.0 SPSS, Munich, Germany). The significance of the differences in cell counts between donor and recipient pairs following HCT were tested using the nonparametric Mann-Whitney U-test. We used special mAb panels to characterize inhibitory receptors on NK and T cells (Table S3, panels 1-4). To identify MHC unrestricted effector cells, we first gated on the NKG2A– LIR1– effector cells and, alternatively, on effector cells that expressed either NKG2A or LIR1. Then, we identified the NK (CD3– CD56+) (panel 1), NKT-like (TCR αβ+ CD56+) (panel 2), γδ T cell (TCR γδ+) (panel 3), and NKT (Vα24+ Vβ11+) (panel 4) cell subsets within this population. Because both NKG2A/CD94 and LIR1 (CD85j) represent inhibitory receptors with broad reactivity against different HLA class I alleles, we designated NKG2A– LIR1– NK cells as “potentially alloreactive.” The NKG2A– LIR1– NK cell subset was further gated on cells which were single-, double-, or triple-positive for KIR2DL1/S1 (CD158a,h), KIR2DL2/L3/S2, (CD158b,j), and KIR3DL1/S1 (CD158e). In addition, we used 4- or 5-color flow cytometry to investigate the expression of activating receptors NKG2D, CD244 (2B4), NKp30, NKp44, and NKp46 on each of these effector cell subsets (Table S3, panels 5-8). Initially, we compared patients before HCT and their healthy donors for the presence of MHC unrestricted killer cells without broadly reactive NK-inhibitory receptors, designated here the NKG2A– LIR1– phenotype (Figure 1A). Only NK cells of the NKG2A– LIR1– phenotype, but not NKG2A– LIR1– NKT-like T cells or γδ T cells, were decreased in the patients before HCT, when compared to the healthy donors (P < .001). In contrast, donors and patients had similar levels of NK cells expressing 1 or both of the broadly reactive NK inhibitory receptors NKG2A and LIR1 (Figure 1B). NKG2A– LIR1– NK cells were already detectable in the donors and became a prominent NK cell subpopulation in the late time course following HCT in some of the patients (Figure 2). There was a relatively sharp rise of NKG2A– LIR1– NK cells between days 28 and 84 post-HCT. Although NKG2A– LIR1– NK cells were, overall, a relatively small population, compared to NK cells expressing NKG2A and/or LIR1, this population continued to rise until approximately day 112 post-HCT, when the levels of the other NK cells were decreasing (Figure 3). As determined by donor KIR genotyping, increased levels of NKG2A– LIR1– NK cells post-HCT were more prominent in patients of the A/B than A/A haplotypes, but there was no apparent difference in the levels of NKG2A+ and/or LIR1+ NK cells (Figure 3). Interestingly, patients remaining in complete remission (CR) had significantly more NKG2A– LIR1– NK cells, but not NKG2A+ and/or LIR1+ NK cells, compared to relapsed patients as observed at 3 independent time points following HCT (Figure 4).Figure 2NKG2A/LIR1 expression on NK cells of representative KIR/HLA-ligand mismatched donor/patient pairs. The NK cells of particular donors and representative patients #28, #30, and #34 were analyzed for NKG2A/LIR1 expression by gating on CD3− CD56+ lymphocytes before HCT (pre Tx) and at different time points following HCT.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Figure 3NK cell levels following HCT analyzed by the donor KIR haplotype. The numbers of (A) NKG2A– LIR1– NK cells and (B) NKG2A+ and/or LIR1+ NK cells, determined from a total of 20,000 effector cells by flow cytometry at different time points following HCT, were analyzed in the donors and their recipients according to the A/B and A/A KIR haplotypes. Patients of the B/B haplotype were not included in this analysis because 5 of 6 B/B patients were KIR-ligand matched.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Figure 4Effector cells positive or negative for NKG2A/LIR1 inhibitory receptors in the posttransplant course. The numbers of NKG2A– LIR1– NK cells as well as NKG2A+ and/or LIR1+ NK cells in patients in CR and in relapse were measured by flow cytometry on days 112, 140, and 168 following HCT. “NKG2A+ and/or LIR1 + NK cells” include the NKG2A+ LIR1–, the LIR1– NKG2A+, and the LIR1+ NKG2A+ subpopulations. The number of patients (n) in both groups at each of these time points is shown on the right. Statistically significant differences (Mann-Whitney U-test) (P < .05) are indicated by ∗ and highly significant differences (P < .001) are indicated by ∗∗.View Large Image Figure ViewerDownload Hi-res image Download (PPT) We monitored NKG2A– LIR1– NK cells from day 14, up to day 168 post-HCT (Figure 5). Assignment of the patients to the “KIR/HLA-ligand match” and “mismatch” groups was performed according to the “KIR/HLA-ligand model” (see the Methods section) [18Hsu K.C. Keever-Taylor C.A. Wilton A. et al.Improved outcome in HLA-identical sibling hematopoietic stem-cell transplantation for acute myelogenous leukemia predicted by KIR and HLA genotypes.Blood. 2005; 105: 4878-4884Crossref PubMed Scopus (412) Google Scholar, 26Leung W. Iyengar R. Triplett B. et al.Comparison of killer Ig-like receptor genotyping and phenotyping for selection of allogeneic blood stem cell donors.J Immunol. 2005; 174: 6540-6545PubMed Google Scholar]. We noticed significantly increased levels of NKG2A– LIR1– NK cells in the blood of the KIR/HLA-ligand mismatched patients compared to the matched patients (Figure 5). This difference was mainly caused by an increase in the amount of NKG2A– LIR1– CD56-dim NK cells (Figure S1a). Nevertheless, the KIR/HLA-ligand mismatched patients also had a tendency toward higher levels of NKG2A– LIR1– CD56-bright NK cells after HCT than the matched patients (Figure S1b). These statistically significant differences were present at several independent time points following HCT, suggesting that these were biologically relevant. Next, we analyzed the NKG2A– LIR1– NK cells in more detail with regard to expression of the relevant KIR, as well as functional capacity using 8-color flow cytometry. NKG2A– LIR1– NK cells expressing KIR for which the patient had no HLA class I ligand (designated KIR-mismatched NK cells) were detectable at relatively low levels before HCT. Following HCT these KIR mismatched NK cells increased in most patients to higher levels than observed in the particular donors with the 2 representative patients shown in Figure 6. Patient #21 had 2 KIR/HLA-ligand mismatches in the presence of an HLA-C group mismatch (donor C1/C2 and patient C2/C2), whereas patient #28 was HLA identical to his donor but still had a KIR3DL1-Bw4 mismatch (Tables 2 and S1). Because the donors for patients #21 and #28 did not carry KIR3DS1, all NK cells expressing KIR3DL1/S1 in these patients following HCT expressed the inhibitory form KIR3DL1. However, patient #21 carried the KIR2DL2/3 and KIR2DS2/3 genes; therefore, both forms of this receptor could be expressed by the single KIR2DL2/L3/S2+ cells in this patient. We examined the functional activity of these potentially alloreactive NK cells by measuring their surface CD107a expression and IFN-γ secretion following stimulation by K562 cells [23Alter G. Malenfant J.M. Altfeld M. CD107a as a functional marker for the identification of natural killer cell activity.J Immunol Methods. 2004; 294: 15-22Crossref PubMed Scopus (1037) Google Scholar, 24Anfossi N. Andre P. Guia S. et al.Human NK cell education by inhibitory receptors for MHC class I.Immunity. 2006; 25: 331-342Abstract Full Text Full Text PDF PubMed Scopus (922) Google Scholar, 25Uhrberg M. The CD107 mobilization assay: viable isolation and immunotherapeutic potential of tumor-cytolytic NK cells.Leukemia. 2005; 19: 707-709Crossref PubMed Scopus (24) Google Scholar]. NKG2A– LIR1– NK cells expressing the mismatched KIRs, as well as other inhibitory KIRs (marked by an arrow in Figure 6A) could clearly be stimulated by K562 cells, revealing their alloreactive potential against target cells that did not express a corresponding inhibitory HLA class I ligand (Figure 6B). In addition, we gated on NKG2A– LIR1– NK cells that expressed only the mismatched KIR but none of the KIR/HLA-ligand matched inhibitory or activating receptors because such NK" @default.
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• W2034287952 cites W1500207187 @default.
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• W2034287952 cites W1969950463 @default.
• W2034287952 cites W1975571037 @default.
• W2034287952 cites W1979849861 @default.
• W2034287952 cites W1989712520 @default.
• W2034287952 cites W1990863238 @default.
• W2034287952 cites W1992908671 @default.
• W2034287952 cites W1994232628 @default.
• W2034287952 cites W2008919922 @default.
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• W2034287952 cites W2011017810 @default.
• W2034287952 cites W2022069273 @default.
• W2034287952 cites W2023144585 @default.
• W2034287952 cites W2025633822 @default.
• W2034287952 cites W2034315146 @default.
• W2034287952 cites W2034544947 @default.
• W2034287952 cites W2036113150 @default.
• W2034287952 cites W2039638023 @default.
• W2034287952 cites W2039805002 @default.
• W2034287952 cites W2040618321 @default.
• W2034287952 cites W2044835282 @default.
• W2034287952 cites W2046294054 @default.
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