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 Blood, 15 June 2004, Vol. 103, No. 12, pp. 4613-4615.
Prepublished online as a Blood First Edition Paper on February 26, 2004; DOI 10.1182/blood-2003-11-3903.

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IMMUNOBIOLOGY
Brief report

Use of B cell–bound HLA-A2 class I monomers to generate high-avidity, allo-restricted CTLs against the leukemia-associated protein Wilms tumor antigen

Philip Savage, Liquan Gao, Kevin Vento, Pam Cowburn, Stephen Man, Neil Steven, Graham Ogg, Andrew McMichael, Agamemnon Epenetos, Els Goulmy, and Hans J. Stauss

From Alexis Biotechnology, London, United Kingdom; Cancer Research Wales, Velindre Hospital, Cardiff, United Kingdom; Department of Immunology, Section of Tumour Immunology, Imperial College, London, United Kingdom; Section of Infection and Immunity, University of Wales College of Medicine, Cardiff, United Kingdom; Division of Cancer Studies, University of Birmingham, Birmingham, United Kingdom; Medical Research Council, Human Immunology Unit, Institute of Molecular Medicine (IMM), Oxford, United Kingdom; Department of Medical Oncology, St Bartholomew's Hospital, London, United Kingdom; and Department of Immunohematology and Blood Transfusion, Leiden University Medical Centre, Leiden, the Netherlands.

Recent studies have detected Wilms tumor antigen (WT1)-specific cytotoxic T lymphocytes (CTLs) in patients with acute myelogenous leukemia (AML) and chronic myelogenous leukemia (CML) and demonstrated that most of these CTLs were low avidity. Although HLA-mismatched donors can mount high-avidity CTLs against HLA-A2-presented peptides of WT1, a dominant anti-alloimmune response usually obscures detection of peptide-specific CTLs. Here we explored the feasibility of using recombinant HLA-A2 monomers containing single peptide epitopes as immunogens to generate peptide-specific CTLs from allogeneic donors. We demonstrate that the coating of HLA-A2- B lymphocytes with A2/peptide monomers provides a strong stimulus for autologous peptide-specific CTLs. After 3 to 5 rounds of stimulation a population of CD8+ T cells binding A2/peptide tetramers is easily detectable by fluorescence-activated cell sorting analysis. Furthermore, sorted A2/WT1 tetramer-positive CTLs display strong cytotoxic activity against leukemia cells expressing WT1 endogenously but not against WT1- human tumor cells. Thus, HLA/peptide monomers may be useful to isolate peptide-specific donor lymphocytes for treatment of patients with leukemia after HLA-mismatched transplantation. (Blood. 2004;103:4613-4615)


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Generating CTL by HLA transplant
Moniek A. de Witte and Ton N. M. Schumacher
Blood 2004 103: 4381-4382. [Full Text] [PDF]



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