title: B-Lymphoma cells with epigenetic silencing of Pax5 trans-differentiate into macrophages, but not other hematopoietic lineages.
abstract: In mice, zygotic or pro-B-cell-specific knock-out of the Pax5 gene allows differentiation of pro-B-cells into all hematopoietic lineages. We previously generated and characterized a murine B-cell lymphoma, dubbed Myc5, whose cells spontaneously lose Pax5 expression when cultured in vitro, but regain it when re-injected into syngeneic mice. In cultured Myc5 cells, the loss of Pax5 correlates with the acquisition of myeloid markers, such as CD11b and F4/80. Here, we sought to determine whether these cells are truly B-macrophage-restricted or, like Pax5-null progenitors, can give rise to additional hematopoietic lineages. In vitro differentiation assays with various cytokines showed that Myc5 cells do not differentiate into NK cells, dendritic cells, neutrophils, or osteoclasts. At the same time, in the presence of macrophage colony-stimulating factor (M-CSF), they readily phagocytose latex beads and provide T-cell help. Both phenomena are indicative of the bona fide macrophage phenotype. Conversely, enforced Pax5 re-expression in macrophage-like Myc5 cells led to down-regulation of the M-CSF receptor and re-acquisition of some B-cell surface markers (e.g., CD79a) and lineage-specific transcription factors (e.g., IRF4 and Blimp). Retrovirally encoded Pax5 also restored expression of several master B-cell differentiation proteins, such as the IL-7 receptor and transcription factor E2A. In contrast, levels of EBF were unaffected by Pax5 suggesting that EBF acts exclusively upstream of Pax5 and might contribute to Pax5 expression. Indeed, transduction with an EBF-encoding retrovirus partly reactivated endogenous Pax5. Our data reveal the complex relationship between B-cell-specific transcription factors and suggest the existence of numerous feedback mechanisms.
authors.affiliation: Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104-4192, USA.
authors.forename: Suchita
authors.initials: S
authors.lastname: Hodawadekar
authors.forename: Duonan
authors.initials: D
authors.lastname: Yu
authors.forename: Diana
authors.initials: D
authors.lastname: Cozma
authors.forename: Bruce
authors.initials: B
authors.lastname: Freedman
authors.forename: Oriol
authors.initials: O
authors.lastname: Sunyer
authors.forename: Michael L
authors.initials: ML
authors.lastname: Atchison
authors.forename: Andrei
authors.initials: A
authors.lastname: Thomas-Tikhonenko
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology*; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D044127:Epigenesis, Genetic; D006412:Hematopoietic Stem Cells / Q000166:cytology; D016393:Lymphoma, B-Cell / Q000235:genetics*; D008264:Macrophages / Q000166:cytology*; D051379:Mice; D051757:PAX5 Transcription Factor / Q000235:genetics*; D010587:Phagocytosis / Q000235:genetics; D013601:T-Lymphocytes / Q000502:physiology; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D015415:Biomarkers; D051757:PAX5 Transcription Factor; C497282:Pax5 protein, mouse