Retrovirus-Mediated Gene Transduction Into Canine Peripheral Blood Repopulating Cells

摘要: Genetically marked peripheral blood progenitor cells were used to investigate their contribution long-term hematopoietic reconstitution after autologous marrow and cell transplantation. After harvest cryopreservation, canine mobilized in three dogs by treatment with recombinant stem factor for 8 days. Peripheral mononuclear collected enriched major histocompatibility complex (MHC) class II antigen-positive avidin-biotin immunoadsorption, thereby enriching repopulating cells. Subsequently, the cocultivated 24 hours on irradiated vector-producing packaging (PA317/LN), followed an 11-day incubation a vector containing culture system. On day of transplantation, animals 9.2 Gy total body irradiation (TBI), transduced untransduced cryopreserved infused within 2 TBI. All engrafted. Two are survivors showing intermittently G418-resistant marrow-derived colony-forming unit granulocyte-macrophage colonies at median 1% 2%, respectively (range, 10%), now up 48 weeks Neo-specific sequences detected polymerase chain reaction granulocytes 65 lymphocytes 75 samples free helper virus no side effects from transduction observed. One died chronic distemper sclerosing encephalitis 84, whereas other two alive 15 17 months. Our data show successful retroviral Long-term persistence myeloid lymphoid transplantation suggests that contains contribute otherwise lethal