title: Exogenous Mtv-7 superantigen transgene expression in major histocompatibility complex class II I-E- mice reconstituted with embryonic stem cell-derived hematopoietic stem cells.
abstract: Direct genetic manipulation of hematopoietic cells is limited by the lack of an established hematopoietic stem cell line. It has been demonstrated that embryonic stem (ES) cell<-->tetraploid embryos are completely ES cell-derived and that fetal liver (FL) cells from these embryos support hematopoiesis in lethally irradiated recipients. In this report, we demonstrate that FL cells from ES cell<-->tetraploid embryos support normal lymphopoiesis and T-cell repertoire development. Moreover, the introduction of the Mtv-7 superantigen transgene coding for minor lymphocyte stimulatory antigen 1 into murine hematopoietic cells via reconstitution with ES cell<-->tetraploid FL cells demonstrates that this method can effectively confer stable genetic changes into the hematopoietic tissues without going through the germ line. Long-term and secondary reconstitution with ES cell<-->tetraploid FL cells expressing the Mtv-7 superantigen transgene clonally deleted minor lymphocyte stimulatory antigen 1-reactive T-cell receptor V beta 6+, -8.1+, and -9+ T cells, but not V beta 7+ T cells, in H-2b (I-E-) mice. This model system will be extremely important for analyzing structure-function relationships of molecules involved in proliferation, differentiation, and selection of hematopoietic cells in vivo and for examining hematopoiesis-specific effects of mutations that are lethal during embryogenesis.
authors.affiliation: Division of Neurobiology and Molecular Immunology, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
authors.forename: C A
authors.initials: CA
authors.lastname: Chambers
authors.forename: J
authors.initials: J
authors.lastname: Kang
authors.forename: J
authors.initials: J
authors.lastname: Pawling
authors.forename: B
authors.initials: B
authors.lastname: Huber
authors.forename: N
authors.initials: N
authors.lastname: Hozumi
authors.forename: A
authors.initials: A
authors.lastname: Nagy
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology; D005260:Female; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000382:microbiology; D000949:Histocompatibility Antigens Class II / Q000235:genetics*; D008099:Liver / Q000166:cytology / Q000196:embryology; D008213:Lymphocyte Activation; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D011123:Polyploidy; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics; D018089:Superantigens / Q000235:genetics*; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D017931:DNA Primers; D000949:Histocompatibility Antigens Class II; C024138:I-E-antigen; D012333:RNA, Messenger; D018089:Superantigens