Article abstract
Nature Methods 6, 520 - 526 (2009)
Published online: 21 June 2009 | doi:10.1038/nmeth.1345
Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers
Sine Reker Hadrup1,4,5, Arnold H Bakker1,4,5, Chengyi J Shu1, Rikke S Andersen2, Jerre van Veluw1, Pleun Hombrink3, Emilie Castermans1, Per thor Straten2, Christian Blank1, John B Haanen1, Mirjam H Heemskerk3 & Ton N Schumacher1
Abstract
The use of fluorescently labeled major histocompatibility complex multimers has become an essential technique for analyzing disease- and therapy-induced T-cell immunity. Whereas classical major histocompatibility complex multimer analyses are well-suited for the detection of immune responses to a few epitopes, limitations on human-subject sample size preclude a comprehensive analysis of T-cell immunity. To address this issue, we developed a combinatorial encoding strategy that allows the parallel detection of a multitude of different T-cell populations in a single sample. Detection of T cells from peripheral blood by combinatorial encoding is as efficient as detection with conventionally labeled multimers but results in a substantially increased sensitivity and, most notably, allows comprehensive screens to be performed. We obtained proof of principle for the feasibility of large-scale screening of human material by analysis of human leukocyte antigen A3–restricted T-cell responses to known and potential melanoma-associated antigens in peripheral blood from individuals with melanoma.
- Division of Immunology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
- Center for Cancer Immune Therapy, Department of Hematology, Herlev University Hospital, Herlev, Denmark.
- Department of Hematology, Leiden University Medical Center, Leiden, The Netherlands.
- Present addresses: Center for Cancer Immune Therapy, Department of Hematology, Herlev University Hospital, Herlev, Denmark (S.R.H.) and Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California, Berkeley, USA (A.H.B.).
- These authors contributed equally to this work.
Correspondence to: Ton N Schumacher1 e-mail: t.schumacher@nki.nl
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