Lymphohematopoietic progenitors immortalized by a retroviral vector harboring a dominant-negative retinoic acid receptor can recapitulate lymphoid, myeloid, and erythroid development.
The lymphohematopoietic progenitors represent < 0.01% of nucleated marrow cells. Here, we describe the immortalization of the murine lymphohematopoietic progenitors by a retroviral vector harboring a dominant-negative retinoic acid receptor. The immortalized progenitors proliferate as a stem-cell-factor-dependent clonal line EML that spontaneously generates pre-pro-B lymphocytes and erythroid and myeloid progenitors. Upon stimulation with interleukin-7 and stromal cells, the pre-pro-B lymphocytes express RAG-1 and undergo D-J rearrangements of the immunoglobulin heavy-chain genes. With erythropoietin the erythroid progenitors proliferate and differentiate into red cells. Generation of the common progenitors for neutrophils and macrophages is suppressed in EML but is inducible by high concentrations of retinoic acid. An additional block in neutrophil differentiation occurs at the promyelocyte stage but can also be overcome by high concentrations of retinoic acid. These studies demonstrate a reproducible way to immortalize lymphohematopoietic progenitors and implicate specific roles for retinoic acid receptors at two distinct stages of hematopoiesis.
Tsai
S
S
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Bartelmez
S
S
Sitnicka
E
E
Collins
S
S
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D001854:Bone Marrow Cells; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002455:Cell Division / Q000187:drug effects; D002461:Cell Line, Transformed; D003433:Crosses, Genetic; D004912:Erythrocytes / Q000166:cytology* / Q000187:drug effects; D004921:Erythropoietin / Q000494:pharmacology; D015321:Gene Rearrangement; D005803:Genes, Immunoglobulin; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D018398:Homeodomain Proteins; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D015851:Interleukin-7 / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008954:Models, Biological; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D018168:Receptors, Retinoic Acid / Q000378:metabolism*; D012190:Retroviridae; D013997:Time Factors; D014212:Tretinoin / Q000494:pharmacology*
D018398:Homeodomain Proteins; D007143:Immunoglobulin Heavy Chains; D015851:Interleukin-7; D012333:RNA, Messenger; D018168:Receptors, Retinoic Acid; D004921:Erythropoietin; C064658:RAG-1 protein; D014212:Tretinoin