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

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.

• Authors: Brigitte Steger, Slavoljub Milosevic, Georg Doessinger, Susanne Reuther, Anja Liepert, Marion Braeu, Julia Schick, Valentin Vogt, Friedhelm Schuster, Tanja Kroell, Dirk H Busch, Arndt Borkhardt, Hans-Jochem Kolb, Johanna Tischer, Raymund Buhmann, Helga Schmetzer
• Journal: Immunobiology (Immunobiology) Vol. 219 Issue 4 Pg. 247-60 (Apr 2014) ISSN: 1878-3279 [Electronic] Netherlands
• PMID: 24315637 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2013 Elsevier GmbH. All rights reserved.

Chemical References

• Antigens, Neoplasm
• Minor Histocompatibility Antigens
• PRAME protein, human
• Peptide Fragments
• WT1 Proteins
• WT1 protein, human
• CD40 Ligand
• Interferon-gamma

Topics

• Antigens, Neoplasm (immunology)
• CD4-Positive T-Lymphocytes (immunology)
• CD40 Ligand (metabolism)
• CD8-Positive T-Lymphocytes (immunology)
• Cell Culture Techniques
• Clone Cells
• Cytotoxicity, Immunologic
• Female
• Graft vs Leukemia Effect (immunology)
• Humans
• Interferon-gamma (metabolism)
• Leukemia, Myeloid, Acute (immunology, therapy)
• Lymphocyte Activation
• Male
• Minor Histocompatibility Antigens (immunology)
• Neoplasm Staging
• Peptide Fragments (immunology)
• Stem Cell Transplantation
• Transplantation, Homologous
• WT1 Proteins (immunology)