Rapid selection of antigen-specific T lymphocytes by retroviral transduction.
Infusions of donor peripheral blood T cells can induce durable remissions of Epstein-Barr virus (EBV) lymphomas complicating marrow grafts, but they contain alloreactive T cells capable of inducing graft-versus-host disease. EBV-specific T-cell lines or clones avoid this problem but require 30 to 40 days of culture to establish. To accelerate the generation of EBV-specific T cells, we tested whether retroviral vectors, which only integrate in dividing cells, could be used to transduce and select antigen-reactive T cells early after sensitization to autologous EBV-transformed B cells. T cells were transduced with a dicistronic retroviral vector, NIT, which encodes low-affinity nerve growth factor receptor as an immunoselectable marker and herpes simplex virus thymidine kinase as a suicide gene, at different time points after sensitization. EBV-specific cytotoxic T lymphocyte precursor (CTLp) frequencies in purified NIT(+) T-cell fractions transduced on day 8 of culture were comparable to those of EBV-specific T-cell lines cultured for 30 days or more. Alloreactive CTLp frequencies were markedly reduced in the NIT(+) fraction relative to the untransduced T-cell population. NIT(+) fractions transduced on day 8 possessed more CD4(+) T cells than the cell lines at day 30 and exhibited the same selective pattern of reactivity against immunodominant antigens presented by specific HLA alleles. In contrast, T cells transduced with NIT 5 days after stimulation with mitogen and interleukin-2 were relatively depleted of T cells specific for autologous EBV-transformed cells. Thus, retroviral vectors may be used for rapid selection of viral antigen-reactive T cells depleted of alloreactive T cells.
Koehne
G
G
Bone Marrow Transplant Service, Department of Pediatrics, Memorial Hospital, New York, NY, USA.
Gallardo
H F
HF
Sadelain
M
M
O'Reilly
R J
RJ
D003602:Cytotoxicity, Immunologic; D005434:Flow Cytometry / Q000379:methods; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006680:HLA Antigens / Q000276:immunology; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D008213:Lymphocyte Activation; D012190:Retroviridae / Q000235:genetics; D013601:T-Lymphocytes / Q000145:classification / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000821:virology; D016662:Virus Integration
D006680:HLA Antigens