Rapid selection of antigen-specific T lymphocytes by retroviral transduction Journal Article
| Authors: | Koehne, G.; Gallardo, H. F.; Sadelain, M.; O'Reilly, R. J. |
| Article Title: | Rapid selection of antigen-specific T lymphocytes by retroviral transduction |
| Abstract: | 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. (C) 2000 by The American Society of Hematology. |
| Keywords: | controlled study; human cell; flow cytometry; binding affinity; t-lymphocytes; gene transfer; genetic vectors; lymphocyte activation; antigen specificity; t-lymphocytes, cytotoxic; lymphoma; cytotoxicity, immunologic; hla antigens; t lymphocyte activation; epstein barr virus; cell selection; retrovirus; retroviridae; herpesvirus 4, human; virus vector; gene transfer techniques; virus integration; humans; human; priority journal; article |
| Journal Title: | Blood |
| Volume: | 96 |
| Issue: | 1 |
| ISSN: | 0006-4971 |
| Publisher: | American Society of Hematology |
| Date Published: | 2000-07-01 |
| Start Page: | 109 |
| End Page: | 117 |
| Language: | English |
| PUBMED: | 10891438 |
| PROVIDER: | scopus |
| DOI: | 10.1182/blood.V96.1.109 |
| DOI/URL: | |
| Notes: | Export Date: 18 November 2015 -- Source: Scopus |
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