2012 Landes Bioscience. Do not distribute. Lymphoblastoid cell line with B1 cell characteristics established from a chronic lymphocytic leukemia clone by in vitro EBV infection

Chronic lymphocytic leukemia (CLL) cells express the receptor for Epstein-Barr virus (EBV) and can be infected in vitro. Infected cells do not express the growth-promoting set of EBV-encoded genes and therefore they do not yield LCLs, in most experiments. With exceptional clones, lines were obtained however. We describe a new line, HG3, established by in vitro EBV-infection from an IGHV1–2 unmutated CLL patient clone. All cells expressed EBNA-2 and LMP-1, the EBV-encoded genes pivotal for transformation. The karyotype, FISH cytogenetics and SNP-array profile of the line and the patient's ex vivo clone showed biallelic 13q14 deletions with genomic loss of DLEU7, miR15a/miR16–1, the two micro-RNAs that are deleted in 50% of CLL cases. Further features of CLL cells were: expression of CD5/CD20/CD27/CD43 and release of IgM natural antibodies reacting with oxLDL-like epitopes on apoptotic cells (cf. stereotyped subset-1). Comparison with two LCLs established from normal B cells showed 32 genes expressed at higher levels (> 2-fold). Among these were LHX2 and LILRA. These genes may play a role in the development of the disease. LHX2 expression was shown in self-renewing multipotent hematopoietic stem cells, and LILRA4 codes for a receptor for bone marrow stromal cell antigen-2 that contributes to B cell development. Twenty-four genes were expressed at lower levels, among these PARD3 that is essential for asymmetric cell division. These genes may contribute to establish precursors of CLL clones by regulation of cellular phenotype in the hematopoietic compartment. Expression of CD5/CD20/CD27/CD43 and spontaneous production of natural antibodies may identify the CLL cell as a self-renewing B1 lymphocyte.

[1]  M. Keating,et al.  The phosphoinositide 3'-kinase delta inhibitor, CAL-101, inhibits B-cell receptor signaling and chemokine networks in chronic lymphocytic leukemia. , 2011, Blood.

[2]  K. Akashi,et al.  Self-renewing hematopoietic stem cell is the primary target in pathogenesis of human chronic lymphocytic leukemia. , 2011, Cancer cell.

[3]  N. Chiorazzi,et al.  Cellular origin(s) of chronic lymphocytic leukemia: cautionary notes and additional considerations and possibilities. , 2011, Blood.

[4]  T. Rothstein,et al.  Human B1 cells in umbilical cord and adult peripheral blood express the novel phenotype CD20+CD27+CD43+CD70− , 2011, The Journal of experimental medicine.

[5]  Richard Sherry,et al.  The lymph node microenvironment promotes B-cell receptor signaling, NF-kappaB activation, and tumor proliferation in chronic lymphocytic leukemia. , 2011, Blood.

[6]  Anurag Tripathi,et al.  2011 Landes Bioscience. Do not distribute. , 2011 .

[7]  P. Youinou,et al.  CD5 expression in B cells from patients with systemic lupus erythematosus. , 2011, Critical reviews in immunology.

[8]  N. Nagy,et al.  Restricted expression of EBV encoded proteins in in vitro infected CLL cells. , 2010, Seminars in cancer biology.

[9]  U. Klein,et al.  New insights into the pathogenesis of chronic lymphocytic leukemia. , 2010, Seminars in cancer biology.

[10]  R. Rosenquist,et al.  Antigens in chronic lymphocytic leukemia--implications for cell origin and leukemogenesis. , 2010, Seminars in cancer biology.

[11]  G. Superti-Furga,et al.  CD14 is a coreceptor of Toll-like receptors 7 and 9 , 2010, The Journal of experimental medicine.

[12]  H. Döhner,et al.  Soluble CD14 is a novel monocyte-derived survival factor for chronic lymphocytic leukemia cells, which is induced by CLL cells in vitro and present at abnormally high levels in vivo. , 2010, Blood.

[13]  N. Chiorazzi,et al.  Many chronic lymphocytic leukemia antibodies recognize apoptotic cells with exposed nonmuscle myosin heavy chain IIA: implications for patient outcome and cell of origin. , 2010, Blood.

[14]  Y. Pekarsky,et al.  13q14 deletions in CLL involve cooperating tumor suppressors. , 2010, Blood.

[15]  M. Rivera,et al.  A genome-wide screen for microdeletions reveals disruption of polarity complex genes in diverse human cancers. , 2010, Cancer research.

[16]  Andrea Califano,et al.  The DLEU2/miR-15a/16-1 cluster controls B cell proliferation and its deletion leads to chronic lymphocytic leukemia. , 2010, Cancer cell.

[17]  L. Lanier,et al.  Regulation of TLR7/9 responses in plasmacytoid dendritic cells by BST2 and ILT7 receptor interaction , 2009, The Journal of experimental medicine.

[18]  L. Lenz.  CD5 sweetens lymphocyte responses , 2009, Proceedings of the National Academy of Sciences.

[19]  J. Yélamos,et al.  The CD5 ectodomain interacts with conserved fungal cell wall components and protects from zymosan-induced septic shock-like syndrome , 2009, Proceedings of the National Academy of Sciences.

[20]  Xiao-Jie Yan,et al.  From bloodjournal.hematologylibrary.org at PENN STATE UNIVERSITY on February 21, 2013. For personal use only. , 2007 .

[21]  E. Meffre,et al.  Chronic Lymphocytic Leukemia Cells Recognize Conserved Epitopes Associated with Apoptosis and Oxidation , 2008, Molecular medicine.

[22]  J. Trowsdale,et al.  The extended human leukocyte receptor complex: diverse ways of modulating immune responses , 2008, Immunological reviews.

[23]  R. Küppers,et al.  EBV transformation overrides gene expression patterns of B cell differentiation stages. , 2008, Molecular immunology.

[24]  A. Hägglund,et al.  Lhx2 Expression Promotes Self-Renewal of a Distinct Multipotential Hematopoietic Progenitor Cell in Embryonic Stem Cell-Derived Embryoid Bodies , 2008, PloS one.

[25]  R. Rosenquist,et al.  A new perspective: molecular motifs on oxidized LDL, apoptotic cells, and bacteria are targets for chronic lymphocytic leukemia antibodies. , 2008, Blood.

[26]  R. Houlston,et al.  Scan of 977 nonsynonymous SNPs in CLL4 trial patients for the identification of genetic variants influencing prognosis. , 2008, Blood.

[27]  Nikolaos Laoutaris,et al.  Stereotyped patterns of somatic hypermutation in subsets of patients with chronic lymphocytic leukemia: implications for the role of antigen selection in leukemogenesis. , 2007, Blood.

[28]  N. Chiorazzi,et al.  CD38 expression labels an activated subset within chronic lymphocytic leukemia clones enriched in proliferating B cells. , 2007, Blood.

[29]  N. Chiorazzi,et al.  CD5+ B cells with the features of subepithelial B cells found in human tonsils , 2007, European journal of immunology.

[30]  C W Caldwell,et al.  Differential DNA methylation patterns of small B-cell lymphoma subclasses with different clinical behavior , 2006, Leukemia.

[31]  Y. Shi,et al.  Immunomodulatory effects of Toll-like receptor-7 activation on chronic lymphocytic leukemia cells , 2006, Leukemia.

[32]  E. Montecino-Rodriguez,et al.  Identification of a B-1 B cell–specified progenitor , 2006, Nature Immunology.

[33]  K. Takada,et al.  Analysis of the transformation of human lymphocytes by Epstein-Barr virus III , 1981, Medical Microbiology and Immunology.

[34]  G. Klein,et al.  EBV Infection Induces Expression of the Transcription Factors ATF-2/c-Jun in B Lymphocytes but not in B-CLL Cells , 2005, Virus Genes.

[35]  J. Kearney.  Innate-like B cells , 2005, Springer Seminars in Immunopathology.

[36]  M. Päivänsalo,et al.  Immunoglobulin M Type of Autoantibodies to Oxidized Low-Density Lipoprotein Has an Inverse Relation to Carotid Artery Atherosclerosis , 2003, Circulation.

[37]  S. Yamada,et al.  Involvement of CD27/CD70 interactions in antigen‐specific cytotoxic T‐lymphocyte (CTL) activity by perforin‐mediated cytotoxicity , 2002, Clinical and experimental immunology.

[38]  C. Croce,et al.  Frequent deletions and down-regulation of micro- RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[39]  John Savill,et al.  Apoptosis disables CD31-mediated cell detachment from phagocytes promoting binding and engulfment , 2002, Nature.

[40]  S. Knuutila,et al.  Cryopreserved chronic lymphocytic leukemia cells analyzed by multicolor fluorescence in situ hybridization after optimized mitogen stimulation , 2002, Genes, chromosomes & cancer.

[41]  J. Yannelli,et al.  Cell surface phenotyping and cytokine production of Epstein-Barr Virus (EBV)-transformed lymphoblastoid cell lines (LCLs). , 2002, Journal of immunological methods.

[42]  Xiao-Jie Yan,et al.  B-cell chronic lymphocytic leukemia cells express a surface membrane phenotype of activated, antigen-experienced B lymphocytes. , 2002, Blood.

[43]  Göran Roos,et al.  Somatically mutated Ig V(H)3-21 genes characterize a new subset of chronic lymphocytic leukemia. , 2002, Blood.

[44]  David Botstein,et al.  Relation of Gene Expression Phenotype to Immunoglobulin Mutation Genotype in B Cell Chronic Lymphocytic Leukemia , 2001, The Journal of experimental medicine.

[45]  Y. Tu,et al.  Gene Expression Profiling of B Cell Chronic Lymphocytic Leukemia Reveals a Homogeneous Phenotype Related to Memory B Cells , 2001, The Journal of experimental medicine.

[46]  H. Meyerson,et al.  CD5 expression by B lymphocytes and its regulation upon Epstein–Barr virus transformation , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[47]  K. Furge,et al.  Gene expression profiling of clear cell renal cell carcinoma: Gene identification and prognostic classification , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[48]  P. Circosta,et al.  MEC1 and MEC2: two new cell lines derived from B-chronic lymphocytic leukaemia in prolymphocytoid transformation. , 1999, Leukemia research.

[49]  S. Anderson,et al.  Epstein-Barr virus LMP2A drives B cell development and survival in the absence of normal B cell receptor signals. , 1998, Immunity.

[50]  A. Devitt,et al.  Human CD14 mediates recognition and phagocytosis of apoptotic cells , 1998, Nature.

[51]  E. Klein,et al.  Recognition of B-CLL cells experimentally infected with EBV by autologous T lymphocytes. , 1998, Immunology letters.

[52]  H. Mellstedt,et al.  CD6 ligation modulates the Bcl-2/Bax ratio and protects chronic lymphocytic leukemia B cells from apoptosis induced by anti-IgM. , 1997, Blood.

[53]  G. Klein,et al.  B‐CLL cells with unusual properties , 1997, International journal of cancer.

[54]  H. Mellstedt,et al.  CD6 ligation modulates BCL-2/BAX ratio and protects B-CLL cells from apoptosis induced by ANTI-IgM , 1996 .

[55]  H. Heng,et al.  Identification of a human LIM-Hox gene, hLH-2, aberrantly expressed in chronic myelogenous leukaemia and located on 9q33-34.1. , 1996, Oncogene.

[56]  T. Kipps,et al.  Expression of CD27 and its ligand, CD70, on chronic lymphocytic leukemia B cells. , 1995, Blood.

[57]  A. Siegbahn,et al.  Epstein-Barr virus (EBV) can immortalize B-cll cells activated by cytokines. , 1994, Leukemia.

[58]  F. Alt,et al.  LH-2: a LIM/homeodomain gene expressed in developing lymphocytes and neural cells. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[59]  L. Florensa,et al.  Immunocytochemical investigation of normal and chronic lymphocytic leukaemia lymphocytes reveals unexpectedly frequent reactivity with some myelomonocytic associated antibodies. , 1992, Leukemia research.

[60]  D. Catovsky,et al.  Activation and immortalization of leukaemic B cells by epstein‐barr virus , 1989, International journal of cancer.

[61]  G. Klein,et al.  EBV‐transformed lymphoblastoid cell lines down‐regulate ebna in parallel with secretory differentiation , 1987, International journal of cancer.

[62]  G. Klein,et al.  Establishment of a lymphoid cell line from leukemic cells of a patient with chronic lymphocytic leukemia , 1980, International journal of cancer.

[63]  G. Klein,et al.  Chromosome analyses of lymphoid cell lines derived from patients with chronic lymphocytic leukemia , 1980, International journal of cancer.

[64]  K. Takada,et al.  Analysis of the transformation of human lymphocytes by Epstein-Barr virus. II. Abortive response of leukemic cells to the transforming virus. , 1980, Intervirology.

[65]  V. Najfeld,et al.  CHRONIC LYMPHOCYTIC LEUKÆMIA: CLONAL ORIGIN IN A COMMITTED B-LYMPHOCYTE PROGENITOR , 1978, The Lancet.