An anti-PR1/HLA-A2 T-cell receptor-like antibody mediates complement-dependent cytotoxicity against acute myeloid leukemia progenitor cells.

PR1 (VLQELNVTV) is a human leukocyte antigen-A2 (HLA-A2)-restricted leukemia-associated peptide from proteinase 3 (P3) and neutrophil elastase (NE) that is recognized by PR1-specific cytotoxic T lymphocytes that contribute to cytogenetic remission of acute myeloid leukemia (AML). We report a novel T-cell receptor (TCR)-like immunoglobulin G2a (IgG2a) antibody (8F4) with high specific binding affinity (dissociation constant [K(D)] = 9.9nM) for a combined epitope of the PR1/HLA-A2 complex. Flow cytometry and confocal microscopy of 8F4-labeled cells showed significantly higher PR1/HLA-A2 expression on AML blasts compared with normal leukocytes (P = .046). 8F4 mediated complement-dependent cytolysis of AML blasts and Lin(-)CD34(+)CD38(-) leukemia stem cells (LSCs) but not normal leukocytes (P < .005). Although PR1 expression was similar on LSCs and hematopoietic stem cells, 8F4 inhibited AML progenitor cell growth, but not normal colony-forming units from healthy donors (P < .05). This study shows that 8F4, a novel TCR-like antibody, binds to a conformational epitope of the PR1/HLA-A2 complex on the cell surface and mediates specific lysis of AML, including LSCs. Therefore, this antibody warrants further study as a novel approach to targeting leukemia-initiating cells in patients with AML.

[1]  M. Talpaz,et al.  PR1-Specific T Cells Are Associated with Unmaintained Cytogenetic Remission of Chronic Myelogenous Leukemia After Interferon Withdrawal , 2010, PloS one.

[2]  J. Weidanz,et al.  TCR Mimic Monoclonal Antibody Targets a Specific Peptide/HLA Class I Complex and Significantly Impedes Tumor Growth In Vivo Using Breast Cancer Models , 2010, The Journal of Immunology.

[3]  P. Hanley,et al.  Functionally active virus-specific T cells that target CMV, adenovirus, and EBV can be expanded from naive T-cell populations in cord blood and will target a range of viral epitopes. , 2009, Blood.

[4]  V. Cerundolo,et al.  Rational development of high-affinity T-cell receptor-like antibodies , 2009, Proceedings of the National Academy of Sciences.

[5]  E. Klechevsky,et al.  Antitumor activity of immunotoxins with T-cell receptor-like specificity against human melanoma xenografts. , 2008, Cancer research.

[6]  I. Pastan,et al.  Targeting TARP, a novel breast and prostate tumor‐associated antigen, with T cell receptor‐like human recombinant antibodies , 2008, European journal of immunology.

[7]  G. Kaiafa,et al.  Detection of CD55- and CD59-deficient granulocytic populations in patients with myelodysplastic syndrome , 2008, Annals of Hematology.

[8]  K. Rezvani,et al.  Leukemia-associated antigen-specific T-cell responses following combined PR1 and WT1 peptide vaccination in patients with myeloid malignancies. , 2007, Blood.

[9]  G. Weiner.  Monoclonal antibody mechanisms of action in cancer , 2007, Immunologic research.

[10]  C. Rinaldo,et al.  Development and implementation of a direct detection, quantitation and validation system for class I MHC self-peptide epitopes. , 2007, Journal of immunological methods.

[11]  S. Wright,et al.  Antibody Targeting to a Class I MHC-Peptide Epitope Promotes Tumor Cell Death , 2006, The Journal of Immunology.

[12]  S. Harding,et al.  A new anticancer glycolipid monoclonal antibody, SC104, which directly induces tumor cell apoptosis. , 2006, Cancer research.

[13]  M. Bonneville,et al.  Assessment of CD8 involvement in T cell clone avidity by direct measurement of HLA‐A2/Mage3 complex density using a high‐affinity TCR like monoclonal antibody , 2005, European journal of immunology.

[14]  M. Peters,et al.  Cellular and complement-dependent cytotoxicity of Ep-CAM-specific monoclonal antibody MT201 against breast cancer cell lines , 2005, British Journal of Cancer.

[15]  M. Pfreundschuh,et al.  Dissecting cytotoxic T cell responses towards the NY‐ESO‐1 protein by peptide/MHC‐specific antibody fragments , 2004, European journal of immunology.

[16]  T. Kyo,et al.  Induction of WT1 (Wilms' tumor gene)-specific cytotoxic T lymphocytes by WT1 peptide vaccine and the resultant cancer regression. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. Dick,et al.  Acute myeloid leukemia originates from a hierarchy of leukemic stem cell classes that differ in self-renewal capacity , 2004, Nature Immunology.

[18]  A. Kennedy,et al.  Three new assays for rituximab based on its immunological activity or antigenic properties: analyses of sera and plasmas of RTX-treated patients with chronic lymphocytic leukemia and other B cell lymphomas. , 2004, Journal of immunological methods.

[19]  D. Metcalfe,et al.  Activation of human mast cells by aggregated IgG through FcgammaRI: additive effects of C3a. , 2004, Clinical immunology.

[20]  Simon J Davis,et al.  Molecular interactions mediating T cell antigen recognition. , 2003, Annual review of immunology.

[21]  A. Gaiger,et al.  WT1 in acute leukemia, chronic myelogenous leukemia and myelodysplastic syndrome: therapeutic potential of WT1 targeted therapies , 2003, Leukemia.

[22]  Mark M. Davis,et al.  Chronic myelogenous leukemia shapes host immunity by selective deletion of high-avidity leukemia-specific T cells. , 2003, The Journal of clinical investigation.

[23]  J. Altman,et al.  MHC‐Peptide Tetramers to Visualize Antigen‐Specific T Cells , 2003, Current protocols in immunology.

[24]  E. Thiel,et al.  CD8 T-cell responses to Wilms tumor gene product WT1 and proteinase 3 in patients with acute myeloid leukemia. , 2002, Blood.

[25]  I. Weissman,et al.  Stem cells, cancer, and cancer stem cells , 2001, Nature.

[26]  H. Jacobs,et al.  TCR-independent T cell development mediated by gain-of-oncogene function or loss-of-tumor-suppressor gene function. , 2000, Seminars in immunology.

[27]  Mark M. Davis,et al.  Evidence that specific T lymphocytes may participate in the elimination of chronic myelogenous leukemia , 2000, Nature Medicine.

[28]  S E Hufton,et al.  Direct selection of a human antibody fragment directed against the tumor T-cell epitope HLA-A1-MAGE-A1 from a nonimmunized phage-Fab library. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[29]  Jan Engberg,et al.  Visualization of Myelin Basic Protein (Mbp) T Cell Epitopes in Multiple Sclerosis Lesions Using a Monoclonal Antibody Specific for the Human Histocompatibility Leukocyte Antigen (Hla)-Dr2–Mbp 85–99 Complex , 2000, The Journal of experimental medicine.

[30]  H. Canpınar,et al.  Expression of complement regulatory proteins CR1, DAF, MCP and CD59in haematological malignancies , 2000, European journal of haematology.

[31]  M. Davis,et al.  A PR1-human leukocyte antigen-A2 tetramer can be used to isolate low-frequency cytotoxic T lymphocytes from healthy donors that selectively lyse chronic myelogenous leukemia. , 1999, Cancer research.

[32]  M. Czuczman,et al.  Association of serum Rituximab (IDEC-C2B8) concentration and anti-tumor response in the treatment of recurrent low-grade or follicular non-Hodgkin's lymphoma. , 1998, Annals of oncology : official journal of the European Society for Medical Oncology.

[33]  A. Barrett,et al.  Cytotoxic T lymphocytes specific for a nonpolymorphic proteinase 3 peptide preferentially inhibit chronic myeloid leukemia colony-forming units. , 1997, Blood.

[34]  J. Hoidal,et al.  Human Proteinase-3 Expression Is Regulated by PU.1 in Conjunction with a Cytidine-rich Element* , 1996, The Journal of Biological Chemistry.

[35]  K. Aozasa,et al.  Increased expression of the Wilms tumor gene (WT1) at relapse in acute leukemia. , 1996, Blood.

[36]  K. Parker,et al.  Targeted T-cell therapy for human leukemia: cytotoxic T lymphocytes specific for a peptide derived from proteinase 3 preferentially lyse human myeloid leukemia cells. , 1996, Blood.

[37]  J. Takeda,et al.  Mechanisms by which the surface expression of the glycosyl-phosphatidylinositol-anchored complement regulatory proteins decay-accelerating factor (CD55) and CD59 is lost in human leukaemia cell lines. , 1996, The Biochemical journal.

[38]  P. S. Andersen,et al.  A recombinant antibody with the antigen-specific, major histocompatibility complex-restricted specificity of T cells. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[39]  U. Gullberg,et al.  The heterogeneity of azurophil granules in neutrophil promyelocytes: immunogold localization of myeloperoxidase, cathepsin G, elastase, proteinase 3, and bactericidal/permeability increasing protein. , 1994, Blood.

[40]  K. Hogan,et al.  Both major and minor peptide-binding pockets in HLA-A2 influence the presentation of influenza virus matrix peptide to cytotoxic T lymphocytes. , 1994, Molecular immunology.

[41]  K. Hogan,et al.  Residues outside of the HLA-A2 peptide-binding groove can abrogate or enhance recognition of influenza virus matrix peptide pulsed cells by cytotoxic T lymphocytes. , 1994, Molecular immunology.

[42]  M. Caligiuri,et al.  A cell initiating human acute myeloid leukaemia after transplantation into SCID mice , 1994, Nature.

[43]  J. Falkenburg,et al.  Recognition of minor histocompatibility antigens on lymphocytic and myeloid leukemic cells by cytotoxic T-cell clones. , 1994, Blood.

[44]  P. Lichter,et al.  Three human elastase-like genes coordinately expressed in the myelomonocyte lineage are organized as a single genetic locus on 19pter. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[45]  S. L. Brown,et al.  Localization and characterization of serologic epitopes on HLA-A2. , 1992, Human immunology.

[46]  M. Bednarek,et al.  The minimum peptide epitope from the influenza virus matrix protein. Extra and intracellular loading of HLA-A2. , 1991, Journal of immunology.

[47]  P. A. Biro,et al.  Molecular characterization of serologic recognition sites in the human HLA-A2 molecule. , 1988, Journal of immunology.

[48]  H. Müller-Eberhard,et al.  Induction of immune cytolysis: tumor-cell killing by complement is initiated by covalent complex of monoclonal antibody and stable C3/C5 convertase. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[49]  P. Parham,et al.  A monoclonal antibody that recognizes an antigenic determinant shared by HLA A2 and B17. , 1980, Human immunology.

[50]  M. Lanotte,et al.  The phylogeny of proteinase 3/myeloblastin, the autoantigen in Wegener's granulomatosis, and myeloperoxidase as shown by immunohistochemical studies on human leukemic cell lines. , 1994, Clinical immunology and immunopathology.