A novel latent membrane 2 transcript expressed in Epstein-Barr virus-positive NK- and T-cell lymphoproliferative disease encodes a target for cellular immunotherapy.

Therapeutic targeting of virus-encoded proteins using cellular immunotherapy has proved successful for Epstein-Barr virus (EBV)-associated posttransplant lymphoproliferative disease. However, the more limited repertoire and immunogenicity of EBV-encoded proteins in other malignancies such as Hodgkin lymphoma and extranodal natural killer (NK)/T lymphoma has been more challenging to target. The immunosubdominant latent membrane protein 2 (LMP2) is considered the optimal target in such Latency II tumors, although data relating to its expression in T/NK malignancies are limited. In addressing the validity of LMP2 as an immunotherapeutic target we found that LMP2-specific effector CD8(+) T cells recognized and killed EBV-positive NK- and T-cell tumor lines, despite an apparent absence of LMP2A protein and barely detectable levels of LMP2 transcripts from the conventional LMP2A and LMP2B promoters. We resolved this paradox by identifying in these lines a novel LMP2 mRNA, initiated from within the EBV terminal repeats and containing downstream, epitope-encoding exons. This same mRNA was also highly expressed in primary (extra-nodal) NK/T lymphoma tissue, with virtually undetectable levels of conventional LMP2A/B transcripts. Expression of this novel transcript in T/NK-cell lymphoproliferative diseases validates LMP2 as an attractive target for cellular immunotherapy and implicates this truncated LMP2 protein in NK- and T-cell lymphomagenesis. This study is registered at clinicaltrials.gov as NCT00062868.

[1]  T. Okamura,et al.  Excellent outcome of allogeneic hematopoietic SCT with reduced-intensity conditioning for the treatment of chronic active EBV infection , 2011, Bone Marrow Transplantation.

[2]  Hao Liu,et al.  Long-term outcome of EBV-specific T-cell infusions to prevent or treat EBV-related lymphoproliferative disease in transplant recipients. , 2010, Blood.

[3]  Y. Nishiyama,et al.  Quantitative analysis of Epstein-Barr virus (EBV)-related gene expression in patients with chronic active EBV infection. , 2010, The Journal of general virology.

[4]  D. Weisenburger,et al.  Clinical differences between nasal and extranasal natural killer/T-cell lymphoma: a study of 136 cases from the International Peripheral T-Cell Lymphoma Project. , 2009, Blood.

[5]  M. Rowe,et al.  Cyclical Expression of EBV Latent Membrane Protein 1 in EBV-Transformed B Cells Underpins Heterogeneity of Epitope Presentation and CD8+ T Cell Recognition1 , 2009, The Journal of Immunology.

[6]  R. deLeeuw,et al.  Genomic analyses reveal global functional alterations that promote tumor growth and novel tumor suppressor genes in natural killer-cell malignancies , 2009, Leukemia.

[7]  M. Ressing,et al.  The Epstein-Barr Virus G-Protein-Coupled Receptor Contributes to Immune Evasion by Targeting MHC Class I Molecules for Degradation , 2009, PLoS pathogens.

[8]  Y. Morishima,et al.  Epstein–Barr virus nuclear antigen 1‐specific CD4+ T cells directly kill Epstein–Barr virus‐carrying natural killer and T cells , 2008, Cancer science.

[9]  E. Hui,et al.  EBV Latent Membrane Proteins (LMPs) 1 and 2 as Immunotherapeutic Targets: LMP-Specific CD4+ Cytotoxic T Cell Recognition of EBV-Transformed B Cell Lines1 , 2008, The Journal of Immunology.

[10]  M. Takahara,et al.  Induction of EBV-latent membrane protein 1-specific MHC class II-restricted T-cell responses against natural killer lymphoma cells. , 2008, Cancer research.

[11]  Chung-Che Chang,et al.  Complete responses of relapsed lymphoma following genetic modification of tumor-antigen presenting cells and T-lymphocyte transfer. , 2007, Blood.

[12]  G. Klein,et al.  Concomitant increase of LMP1 and CD25 (IL‐2‐receptor α) expression induced by IL‐10 in the EBV‐positive NK lines SNK6 and KAI3 , 2006, International journal of cancer.

[13]  R. Longnecker,et al.  Epstein-Barr Virus Latent Membrane Protein 2B (LMP2B) Modulates LMP2A Activity , 2006, Journal of Virology.

[14]  E. Hui,et al.  Analysis of Epstein-Barr virus latent gene expression in endemic Burkitt's lymphoma and nasopharyngeal carcinoma tumour cells by using quantitative real-time PCR assays. , 2006, The Journal of general virology.

[15]  Y. Morishima,et al.  Epstein‐Barr virus (EBV) latent membrane protein‐1‐specific cytotoxic T lymphocytes targeting EBV‐carrying natural killer cell malignancies , 2006, European journal of immunology.

[16]  Shulian Wang,et al.  Radiotherapy as primary treatment for stage IE and IIE nasal natural killer/T-cell lymphoma. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[17]  A. Davis,et al.  The Generation and Characterization of LMP2-Specific CTLs for Use as Adoptive Transfer From Patients With Relapsed EBV-Positive Hodgkin Disease , 2004, Journal of immunotherapy.

[18]  G. Ott,et al.  Expression of the Epstein–Barr virus (EBV)‐encoded latent membrane protein 2A (LMP2A) in EBV‐associated nasopharyngeal carcinoma , 2004, The Journal of pathology.

[19]  Ji Yeun Kim,et al.  Therapeutic Outcome of Extranodal NK/T-Cell Lymphoma Initially Treated with Chemotherapy Result of Chemotherapy in NK/T-Cell Lymphoma , 2003, Acta oncologica.

[20]  T. Blankenstein,et al.  Retroviral vectors for high-level transgene expression in T lymphocytes. , 2003, Human gene therapy.

[21]  T. Morio,et al.  Common cytological and cytogenetic features of Epstein‐Barr virus (EBV)‐positive natural killer (NK) cells and cell lines derived from patients with nasal T/NK‐cell lymphomas, chronic active EBV infection and hydroa vacciniforme‐like eruptions , 2003, British journal of haematology.

[22]  A. Davis,et al.  Generating CTLs against the subdominant Epstein-Barr virus LMP1 antigen for the adoptive immunotherapy of EBV-associated malignancies. , 2003, Blood.

[23]  V. Cerundolo,et al.  Identification of a TAP-Independent, Immunoproteasome-Dependent CD8+ T-Cell Epitope in Epstein-Barr Virus Latent Membrane Protein 2 , 2003, Journal of Virology.

[24]  G. Reynolds,et al.  Interleukin 6 expression by Hodgkin/Reed–Sternberg cells is associated with the presence of ‘B’ symptoms and failure to achieve complete remission in patients with advanced Hodgkin's disease , 2002, British journal of haematology.

[25]  Y. Sakiyama,et al.  Differential cellular targets of Epstein-Barr virus (EBV) infection between acute EBV-associated hemophagocytic lymphohistiocytosis and chronic active EBV infection. , 2001, Blood.

[26]  H. Kanegane,et al.  Clinical and virologic characteristics of chronic active Epstein-Barr virus infection. , 2001, Blood.

[27]  T. Sekine,et al.  Characterization of novel natural killer (NK)-cell and gammadelta T-cell lines established from primary lesions of nasal T/NK-cell lymphomas associated with the Epstein-Barr virus. , 2001, Blood.

[28]  D. Srivastava,et al.  Infusion of cytotoxic T cells for the prevention and treatment of Epstein-Barr virus-induced lymphoma in allogeneic transplant recipients. , 1998, Blood.

[29]  S Kimbrough,et al.  Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes for the treatment of patients with EBV-positive relapsed Hodgkin's disease. , 1998, Blood.

[30]  E. Kremmer,et al.  Immunohistochemical Detection of the Epstein-Barr Virus–Encoded Latent Membrane Protein 2A in Hodgkin's Disease and Infectious Mononucleosis , 1997 .

[31]  A. Rickinson,et al.  Conserved CTL epitopes within EBV latent membrane protein 2: a potential target for CTL-based tumor therapy. , 1997, Journal of immunology.

[32]  G. Srivastava,et al.  Nasal NK‐ and T‐cell lymphomas share the same type of Epstein‐Barr virus latency as nasopharyngeal carcinoma and Hodgkin's disease , 1996, International journal of cancer.

[33]  E. Kremmer,et al.  Identification of latent membrane protein 2A (LMP2A) domains essential for the LMP2A dominant-negative effect on B-lymphocyte surface immunoglobulin signal transduction , 1996, Journal of virology.

[34]  N. Raab-Traub,et al.  The Epstein-Barr virus 3.5-kilobase latent membrane protein 1 mRNA initiates from a TATA-Less promoter within the first terminal repeat , 1995, Journal of virology.

[35]  J. Trowsdale,et al.  Restoration of endogenous antigen processing in Burkitt's lymphoma cells by Epstein‐Barr virus latent membrane protein‐1: coordinate up‐regulation of peptide transporters and HLA‐class I antigen expression , 1995, European journal of immunology.

[36]  T. Tursz,et al.  Sequence polymorphism in the Epstein-Barr virus latent membrane protein (LMP)-2 gene. , 1995, The Journal of general virology.

[37]  E. Kremmer,et al.  The Epstein‐Barr virus nuclear antigen 2 interacts with an EBNA2 responsive cis‐element of the terminal protein 1 gene promoter. , 1993, The EMBO journal.

[38]  A. Rickinson,et al.  Three pathways of Epstein-Barr virus gene activation from EBNA1-positive latency in B lymphocytes , 1992, Journal of virology.

[39]  U. Nater,et al.  Epstein-Barr virus. , 1991, The Journal of family practice.

[40]  D. Choy,et al.  PRESENCE OF EPSTEIN‐BARR VIRUS DNA IN NASAL LYMPHOMAS OF B AND ‘T’ CELL TYPE , 1990, Hematological oncology.

[41]  P. Farrell,et al.  The terminal protein gene 2 of Epstein-Barr virus is transcribed from a bidirectional latent promoter region. , 1989, The Journal of general virology.

[42]  E. Kieff,et al.  Expression of Epstein-Barr virus transformation-associated genes in tissues of patients with EBV lymphoproliferative disease. , 1989, The New England journal of medicine.

[43]  E. Kieff,et al.  Two related Epstein-Barr virus membrane proteins are encoded by separate genes , 1989, Journal of virology.

[44]  M. Perricaudet,et al.  A spliced Epstein‐Barr virus gene expressed in immortalized lymphocytes is created by circularization of the linear viral genome. , 1988, The EMBO journal.

[45]  E. Kieff,et al.  Monoclonal antibodies to the latent membrane protein of Epstein-Barr virus reveal heterogeneity of the protein and inducible expression in virus-transformed cells. , 1987, The Journal of general virology.

[46]  I. Mcconnell,et al.  A monoclonal antibody to the HLA-DR product recognizes a polymorphic Ia determinant in mice. , 1981, Immunology.

[47]  C. Barnstable,et al.  Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens-new tools for genetic analysis , 1978, Cell.