Conditional ablation of T-cell development by a novel viral ion channel transgene.
A novel conditional-lethal transgene system is defined in which a mutated influenza A virus ion-channel protein, which is permeable to monovalent cations, is lethal to cells on heterotypic expression and whose activity can be blocked by an antiviral drug (amantadine), is used to reversibly disrupt T-cell development. In vivo expression of the M2 ion channel, as a transgene under control of the T-cell specific p56(Lck) proximal promoter, resulted in total ablation of T-cell development with the accumulation of three distinct populations of early progenitor cells (CD44(+) CD25(-); CD44(+) CD25(+); CD44(+) CD25(hi)) in the thymic rudiment. In vitro development of transgenic fetal thymic progenitors to single-positive T cells could be rescued by antiviral drug treatment. Moreover, there was a radical reduction in B-cell lymphopoiesis, evident at the pre-B-cell stage, with a twofold increase of lymphoid cells 'in cycle' in transgenic bone marrow, indicative of major changes in haematopoietic homeostasis. This system may provide a generic protocol for conditional, lineage-specific cell ablation with available tissue-specific promoters for any eukaryotic developmental system, and provide a window on early T-cell development.
Smith
Claire A
CA
National Institute for Medical Research, The Ridgeway, Mill Hill, London, UK.
Graham
Christine M
CM
Mathers
Kathleen
K
Skinner
Anita
A
Hay
Alan J
AJ
Schroeder
Cornelia
C
Thomas
D Brian
DB
D000547:Amantadine / Q000494:pharmacology; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D001402:B-Lymphocytes / Q000276:immunology; D002453:Cell Cycle / Q000276:immunology; D005804:Genes, Lethal / Q000276:immunology*; D018960:Hyaluronan Receptors / Q000032:analysis; D016130:Immunophenotyping; D009980:Influenza A virus / Q000235:genetics*; D015640:Ion Channel Gating / Q000187:drug effects; D007473:Ion Channels / Q000187:drug effects / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D009924:Organ Culture Techniques; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D015375:Receptors, Interleukin-2 / Q000032:analysis; D013601:T-Lymphocytes / Q000276:immunology*; D013945:Thymoma / Q000276:immunology; D013953:Thymus Neoplasms / Q000276:immunology; D014162:Transfection; D019076:Transgenes / Q000276:immunology*; D014763:Viral Matrix Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D000086082:Viroporin Proteins
D000547:Amantadine; D000998:Antiviral Agents; D018960:Hyaluronan Receptors; D007473:Ion Channels; D019253:Proto-Oncogene Proteins c-bcl-2; D019009:Proto-Oncogene Proteins c-kit; D015375:Receptors, Interleukin-2; D014763:Viral Matrix Proteins; C031540:M-protein, influenza virus; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins