Generation of T-cell lines to autologous acute myeloid leukemia cells by competitive limiting dilution culture of acute myeloid leukemia mononuclear cells.
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.


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.


CONCLUSIONS
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.
Zhong
Rui-Kun
RK
Departments of Medicine and the Moores UCSD Cancer Center, University of California San Diego, La Jolla, Calif. 92093-0960, USA.
Lane
Thomas A
TA
Ball
Edward D
ED
D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000494:pharmacology; D003713:Dendritic Cells / Q000276:immunology; D005434:Flow Cytometry; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D014182:Transplantation, Autologous
D016207:Cytokines