Lymphocytes as cellular vehicles for gene therapy in mouse and man.

Abstract

The application of bone marrow gene therapy has been stalled by the inability to achieve stable high-level gene transfer and expression in the totipotent stem cells. We show that retroviral vectors can stably introduce genes into antigenspecific murine and human T lymphocytes in culture. Murine helper T cells were transduced with the retroviral vector SAX to express both neomycin-resistance and human adenosine deaminase genes. These cells were expanded in culture and selected for expression of neomycin resistance with G418.

A neovascularized organoid derived from retrovirally engineered bone marrow stroma leads to prolonged in vivo systemic delivery of erythropoietin in nonmyeloablated, immunocompetent mice

“…[60][61][62] Hematopoietic stem cells can be difficult to expand in culture and gene-modify, 63,64 and very large numbers are required for engraftment in the absence of a toxic 'conditioning' regimen. 65 Lymphocytes possess a short lifespan, 66 and human umbilical vein endothelial cells cannot be obtained from an adult.…”

Manipulation of MHC-I/TCR Interaction for Immune Therapy

“…There are several reasons to promote the development of adoptive T cell immunotherapy: 1) The method of in vitro culturing specific T cells had been developed (105,112); 2) High avidity CTLs have superior antitumor efficacy in vitro and in vivo (113); 3) Gene delivery vectors are developed to transduce lymphocytes, such as retrovirus vector (114)(115)(116) and lentivector (117); 4) Adoptive T cell therapy overcomes the difficulty of isolating antigen-specific T lymphocytes for individual patients and could break the tolerance of self over-expressed peptides in tumor cell (118,119).…”

β-Galactosidase marker genes to tag and track human hematopoietic cells

“…Various coding sequences including neoR, 122 herpes simplex virus thymidine kinase, 123,124 human CD4, 125 and a truncated version of the human low-affinity nerve growth factor receptor, 73,124 were retrovirally transduced into T cells derived from peripheral blood 124,126 or resected tumors. 127 The culture and transduction of cells require stimulation, which can be performed with a combination of IL-2 and phytohemagglutinin [123][124][125]128,129 or with a combination of antibodies directed against CD3, CD2, or CD28. 126 Retroviral transduction of the ␤-gal activity encoded by various genetic forms of the LacZ gene, including the regular LacZ, the nls-LacZ gene, and the Sh-ble::LacZ fusion gene has been successfully tested in prestimulated human T lymphocytes derived from peripheral blood cultivated in the presence or absence of feeder cells.…”