Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers

TY - JOUR

T1 - Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers

AU - Hadrup, Sine Reker

AU - Bakker, Arnold H

AU - Shu, Chengyi J

AU - Andersen, Rikke S

AU - van Veluw, Jerre

AU - Hombrink, Pleun

AU - Castermans, Emilie

AU - Thor Straten, Per

AU - Blank, Christian

AU - Haanen, John B

AU - Heemskerk, Mirjam H

AU - Schumacher, Ton N

N1 - Keywords: Antigens; Antigens, Neoplasm; Cell Separation; Epitopes; Fluorescent Dyes; Histocompatibility Antigens; Humans; Immunologic Techniques; Nanotechnology; Neoplasm Proteins; Peptides; Protein Structure, Quaternary; Quantum Dots; Sensitivity and Specificity; T-Lymphocyte Subsets

PY - 2009

Y1 - 2009

N2 - 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.

AB - 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.

U2 - 10.1038/nmeth.1345

DO - 10.1038/nmeth.1345

M3 - Journal article

C2 - 19543285

SN - 1548-7091

VL - 6

SP - 520

EP - 526

JO - Nature Methods

JF - Nature Methods

IS - 7

ER -