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

Sine Reker Hadrup, Arnold H. Bakker, Chengyi J. Shu, Rikke S. Andersen, Jerre van Veluw, Pleun Hombrink, Emilie Castermans, Per thor Straten, Christian Blank, John B. Haanen, Mirjam H. Heemskerk, Ton N. Schumacher

Research output: Contribution to journalJournal articleResearchpeer-review

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.
Original languageEnglish
JournalNature Methods
Volume6
Issue number7
Pages (from-to)520-526
ISSN1548-7091
DOIs
Publication statusPublished - 2009
Externally publishedYes

Keywords

  • Biotechnology
  • Molecular Biology
  • Biochemistry
  • Cell Biology
  • allophycocyanin
  • epitope
  • fluorochrome
  • gamma interferon
  • HLA A3 antigen
  • leukocyte antigen
  • major histocompatibility antigen
  • melanoma antigen
  • phycoerythrin
  • polymer
  • quantum dot
  • antigen detection
  • antigen specificity
  • article
  • blood
  • CD8+ T lymphocyte
  • cellular immunity
  • controlled study
  • cytokine release
  • Cytomegalovirus
  • flow cytometry
  • Hepatitis C virus
  • human
  • human cell
  • Human immunodeficiency virus
  • melanoma
  • priority journal
  • T lymphocyte
  • T lymphocyte subpopulation
  • 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
  • T cells
  • Melanoma-Specific Antigens
  • BIOCHEMICAL
  • CLASS-I LIGANDS
  • EPITOPES
  • IDENTIFICATION
  • LYMPHOCYTES
  • TECHNOLOGY
  • COMPLEXES
  • PHENOTYPE
  • MOLECULES
  • EXCHANGE
  • HLA-A1
  • Primates Mammalia Vertebrata Chordata Animalia (Animals, Chordates, Humans, Mammals, Primates, Vertebrates) - Hominidae [86215] human common
  • epitopes
  • human leukocyte antigen A3
  • major histocompatibility complex
  • melanoma-associated antigen
  • 02506, Cytology - Animal
  • 02508, Cytology - Human
  • 10060, Biochemistry studies - General
  • 10064, Biochemistry studies - Proteins, peptides and amino acids
  • 15002, Blood - Blood and lymph studies
  • 15004, Blood - Blood cell studies
  • 18504, Integumentary system - Physiology and biochemistry
  • 18506, Integumentary system - Pathology
  • 24003, Neoplasms - Immunology
  • 24004, Neoplasms - Pathology, clinical aspects and systemic effects
  • 34502, Immunology - General and methods
  • 34508, Immunology - Immunopathology, tissue immunology
  • Biochemistry and Molecular Biophysics
  • Blood and Lymphatics
  • Immune System
  • Integumentary System
  • Methods and Techniques
  • Tumor Biology
  • melanoma Melanoma (MeSH) neoplastic disease, integumentary system disease
  • immune response
  • T-cell immunity
  • T-cell response
  • Chemical Coordination and Homeostasis
  • Transport and Circulation
  • peripheral blood blood and lymphatics
  • T-cell immune system, blood and lymphatics
  • multidimensional encoding laboratory techniques
  • multimer analysis laboratory techniques

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