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Immunobiology. 2014 Apr;219(4):247-60. doi: 10.1016/j.imbio.2013.10.008. Epub 2013 Oct 27.

CD4(+)and CD8(+)T-cell reactions against leukemia-associated- or minor-histocompatibility-antigens in AML-patients after allogeneic SCT.

Author information

1
Helmholtz Center Munich (German Research Center for Environmental Health and Clinical Cooperative Group Hematopoetic Cell-Transplantation), 81377 Munich, Germany; University Hospital of Munich, Department for Hematopoetic Cell Transplantation, Med. Dept. 3, 81377 Munich, Germany.
2
Helmholtz Center Munich (German Research Center for Environmental Health and Clinical Cooperative Group Hematopoetic Cell-Transplantation), 81377 Munich, Germany.
3
Institute for Medical Microbiology, Immunology and Hygiene, and Focus Group'Clinical Cell Processing and Purification', Institute for Advanced Study, Technical University Munich, 81675 Munich, Germany.
4
Department of Pediatric Oncology, Hematology and Immunology, University Hospital Duesseldorf, 40225 Duesseldorf, Germany.
5
University Hospital of Munich, Department for Hematopoetic Cell Transplantation, Med. Dept. 3, 81377 Munich, Germany.
6
Institute for Medical Microbiology, Immunology and Hygiene, and Focus Group'Clinical Cell Processing and Purification', Institute for Advanced Study, Technical University Munich, 81675 Munich, Germany; Clinical Cooperation Groups "Antigen-specific Immunotherapy" and "Immune Monitoring", Helmholtz Center Munich and Technical University Munich, 81675 Munich, Germany.
7
Helmholtz Center Munich (German Research Center for Environmental Health and Clinical Cooperative Group Hematopoetic Cell-Transplantation), 81377 Munich, Germany; University Hospital of Munich, Department for Hematopoetic Cell Transplantation, Med. Dept. 3, 81377 Munich, Germany. Electronic address: Helga.Schmetzer@med.uni-muenchen.de.

Abstract

T-cells play an important role in the remission-maintenance in AML-patients (pts) after SCT, however the role of LAA- (WT1, PR1, PRAME) or minor-histocompatibility (mHag, HA1) antigen-specific CD4(+) and CD8(+)T-cells is not defined. A LAA/HA1-peptide/protein stimulation, cloning and monitoring strategy for specific CD8(+)/CD4(+)T-cells in AML-pts after SCT is given. Our results show that (1) LAA-peptide-specific CD8+T-cells are detectable in every AML-pt after SCT. CD8(+)T-cells, recognizing two different antigens detectable in 5 of 7 cases correlate with long-lasting remissions. Clonal TCR-Vβ-restriction exemplarily proven by spectratyping in PRAME-specific CD8(+)T-cells; high PRAME-peptide-reactivity was CD4(+)-associated, as shown by IFN-γ-release. (2) Two types of antigen-presenting cells (APCs) were tested for presentation of LAA/HA1-proteins to CD4(+)T-cells: miniEBV-transduced lymphoblastoid cells (B-cell-source) and CD4-depleted MNC (source for B-cell/monocyte/DC). We provide a refined cloning-system for proliferating, CD40L(+)CD4(+)T-cells after LAA/HA1-stimulation. CD4(+)T-cells produced cytokines (GM-CSF, IFN-γ) upon exposure to LAA/HA1-stimulation until after at least 7 restimulations and demonstrated cytotoxic activity against naive blasts, but not fibroblasts. Antileukemic activity of unstimulated, stimulated or cloned CD4(+)T-cells correlated with defined T-cell-subtypes and the clinical course of the disease. In conclusion we provide immunological tools to enrich and monitor LAA/HA1-CD4(+)- and CD8(+)T-cells in AML-pts after SCT and generate data with relevant prognostic value. We were able to demonstrate the presence of LAA-peptide-specific CD8(+)T-cell clones in AML-pts after SCT. In addition, we were also able to enrich specific antileukemic reactive CD4(+)T-cells without GvH-reactivity upon repeated LAA/HA1-protein stimulation and limiting dilution cloning.

KEYWORDS:

Acute myeloid leukemia; CD4/CD8 T-cells; Dendritic cells; Immunophenotyping; Minor antigens; Multimer; TAA

PMID:
24315637
DOI:
10.1016/j.imbio.2013.10.008
[Indexed for MEDLINE]

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