title: Stable nontumorigenic phenotype of somatic cell hybrids between malignant Burkitt's lymphoma cells and autologous EBV-immortalized B cells despite induction of chromosomal breakage and loss.
abstract: Fusion of the highly tumorigenic Burkitt's lymphoma (BL) cell line BL60-P7 with the nontumorigenic autologous EBV-immortalized lymphoblastoid cell line (LCL) IARC 277 results in suppression of the tumorigenic phenotype of the parental cell line BL60-P7 after s.c. inoculation into T cell-deficient nude mice. We analyzed whether, after long-term cultivation of these lymphoma hybrid cells, expression of tumorigenicity could be observed and correlated to the loss of particular chromosomes or chromosomal fragments, akin to numerous nonlymphoid hybrid cell models described previously. Two years after fusion, in vitro proliferation of some BL x LCL hybrid cells accelerated, and they partially lost LCL-typical aggregation. However, no major changes in the expression pattern of B cell-associated surface antigens and the EBV latent membrane protein LMP 1 were observed. Cytogenetic evaluation of these cells revealed spontaneous loss of chromosomes. Karyotyping of long-term cultivated hybrid cells demonstrated the occurrence of disomy for each chromosome in at least one metaphase analyzed. Therefore, if suppression of tumorigenicity in these hybrid cells would have been the result of the presence of a single LCL-derived chromosome, there should have been a high probability of its loss, leading to tumorigenic segregants. Surprisingly, the tumorigenic phenotype remained suppressed in nude mice. To induce chromosomal breakage and maldistribution, in addition to spontaneous chromosomal loss, the hybrid cell lines were irradiated at various doses. Again, none of the hybrid cell clones treated in this manner became tumorigenic in nude mice. Immunohistological analysis of the regressing hybrid cell tumors revealed that the hybrid cells had retained their LCL-like differentiation phenotype in vivo. In addition, infiltration with mononuclear cells of murine origin was observed in these regressing hybrid grafts. We conclude that suppression of the tumorigenic Burkitt's lymphoma phenotype in these hybrid cells cannot be attributed to a function encoded by a distinct chromosome or chromosomal fragment. Rather, the unexpected stable nontumorigenic phenotype reflects a LCL-specific activated B-cell phenotype of these hybrids, most probably induced by the expression of numerous copies of episomal latent EBV proteins.
authors.affiliation: Department of Internal Medicine I, University of Cologne, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Jox
authors.forename: E
authors.initials: E
authors.lastname: Taquia
authors.forename: M
authors.initials: M
authors.lastname: Vockerodt
authors.forename: A
authors.initials: A
authors.lastname: Draube
authors.forename: M
authors.initials: M
authors.lastname: Pawlita
authors.forename: P
authors.initials: P
authors.lastname: Möller
authors.forename: J
authors.initials: J
authors.lastname: Bullerdiek
authors.forename: V
authors.initials: V
authors.lastname: Diehl
authors.forename: J
authors.initials: J
authors.lastname: Wolf
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology / Q000648:ultrastructure*; D002051:Burkitt Lymphoma / Q000235:genetics* / Q000276:immunology / Q000473:pathology; D002459:Cell Fusion; D002460:Cell Line; D002869:Chromosome Aberrations; D005260:Female; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D006822:Hybrid Cells / Q000648:ultrastructure*; D008213:Lymphocyte Activation; D051379:Mice; D008819:Mice, Nude; D010641:Phenotype; D014763:Viral Matrix Proteins / Q000235:genetics
chemical_list: C037662:EBV-associated membrane antigen, Epstein-Barr virus; D014763:Viral Matrix Proteins