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  • Title: Generation of T-cell lines to autologous acute myeloid leukemia cells by competitive limiting dilution culture of acute myeloid leukemia mononuclear cells.
    Author: Zhong RK, Lane TA, Ball ED.
    Journal: Exp Hematol; 2008 Apr; 36(4):486-94. PubMed ID: 18249062.
    Abstract:
    OBJECTIVE: To develop a novel method of generating multiple autologous acute myeloid leukemia (AML) reactive T-cell lines as a step toward adoptive immunotherapy for AML. MATERIALS AND METHODS: AML peripheral blood mononuclear cells (MNC), including >90% AML blasts and 1% to 3% T cells, were seeded in limiting dilution culture in which AML blasts were induced to undergo dendritic cell (DC) differentiation. T cells were primed and activated with the addition of a cytokine combination. RESULTS: Highly reactive anti-AML T-cell lines (both CD4(+) and CD8(+)) were generated, selected, and expanded. The estimated average frequency of AML-reactive T cells or precursors was 6 +/- 3/1,000,000 AML peripheral blood mononuclear cells (n = 11). Robust intracellular interferon-gamma (IFN-gamma) release from T-cell lines was demonstrated by flow cytometry after stimulation by autologous AML cells, but not an autologous B-lymphoblastoid cell line (LCL). These T-cell lines caused specific lysis of autologous AML cells, but not autologous LCL or allogeneic AML cells, and they depleted autologous AML colony-forming cells (CFC), but not normal CFC. Most CD4(+) T-cell lines exerted strong proapoptotic effects on AML cells. AML cell apoptosis by CD4(+) T-cell lines correlated with IFN-gamma secretion. CONCLUSION: This study demonstrates a methodology for generating large numbers of AML-reactive cytotoxic T cell lines (either class I or II restricted) that may be useful clinically in adoptive immunotherapy. This study also provides estimates of AML-reactive T-cell frequency in patients with AML.
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