Modular Nucleic Acid Assembled p/MHC Microarrays for Multiplexed Sorting of Antigen-Specific T Cells

Gabriel A. Kwong, Caius G. Radu, Kiwook Hwang, Chengyi J. Shu, Chao Ma, Richard C. Koya, Begonya Comin-Anduix, Sine Reker Hadrup, Ryan C. Bailey, Owen N. Witte, Ton N. Schumacher, Antoni Ribas, James R. Heath

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

The human immune system consists of a large number of T cells capable of recognizing and responding to antigens derived from various sources. The development of peptide-major histocompatibility (p/MHC) tetrameric complexes has enabled the direct detection of these antigen-specific T cells. With the goal of increasing throughput and multiplexing of T cell detection, protein microarrays spotted with defined p/MHC complexes have been reported, but studies have been limited due to the inherent instability and reproducibility of arrays produced via conventional spotted methods. Herein, we report on a platform for the detection of antigen-specific T cells on glass substrates that otters significant advantages over existing surface-bound schemes. In this approach, called "Nucleic Acid Cell Sorting (NACS)", single-stranded DNA oligomers conjugated site-specifically to p/MHC tetramers are employed to immobilize p/MHC tetramers via hybridization to a complementary-printed substrate. Fully assembled p/MHC arrays are used to detect and enumerate T cells captured from cellular suspensions, including primary human T cells collected from cancer patients. NACS arrays outperform conventional spotted arrays assessed in key criteria such as repeatability and homogeneity. The versatility of employing DNA sequences for cell sorting is exploited to enable the programmed, selective release of target populations of immobilized T cells with restriction endonucleases for downstream analysis. Because of the performance, facile and modular assembly of p/MHC tetramer arrays, NACS holds promise as a versatile platform for multiplexed T cell detection.
Original languageEnglish
JournalJournal of the American Chemical Society
Volume131
Issue number28
Pages (from-to)9695-9703
Number of pages9
ISSN0002-7863
DOIs
Publication statusPublished - 2009
Externally publishedYes

Keywords

  • Chemistry (all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry
  • Cancer patients
  • Cell sorting
  • Direct detection
  • Glass substrates
  • Human immune systems
  • Inherent instability
  • Modular assembly
  • Primary human T cells
  • Printed substrate
  • Protein microarray
  • Reproducibility
  • Restriction endonucleases
  • Single-stranded DNA
  • T cells
  • Tetramer
  • Tetrameric complex
  • Tetramers
  • Amines
  • Antigens
  • Bioassay
  • Biochips
  • Cell membranes
  • Cellular arrays
  • DNA
  • DNA sequences
  • Genes
  • Multiplexing
  • Nucleic acids
  • Oligomers
  • Substrates
  • Cytology
  • antigen
  • oligomer
  • peptide
  • restriction endonuclease
  • single stranded DNA
  • tetramer
  • antigen specificity
  • article
  • cancer patient
  • cell selection
  • cell suspension
  • conjugation
  • DNA hybridization
  • DNA microarray
  • DNA sequence
  • human
  • human cell
  • immobilization
  • immobilized cell
  • major histocompatibility complex
  • surface property
  • T lymphocyte
  • Animals
  • Base Sequence
  • Cell Line
  • Cell Separation
  • Cytological Techniques
  • DNA Restriction Enzymes
  • DNA, Complementary
  • DNA, Single-Stranded
  • Glass
  • Histocompatibility Antigens
  • Humans
  • Immobilized Proteins
  • Mice
  • Nucleic Acid Hybridization
  • Protein Array Analysis
  • Protein Multimerization
  • Protein Structure, Quaternary
  • Receptors, Antigen, T-Cell
  • Reproducibility of Results
  • Streptavidin
  • Substrate Specificity
  • T-Lymphocytes
  • 9013-20-1 Streptavidin
  • EC 3.1.21.- DNA Restriction Enzymes
  • ANTIGENS
  • CHEMISTRY,
  • CLASS-I LIGANDS
  • ENGINEERED LYMPHOCYTES
  • CANCER REGRESSION
  • ESCHERICHIA-COLI
  • MELANOMA ANTIGEN
  • FLOW-CYTOMETRY
  • MHC TETRAMERS
  • IMMOBILIZATION
  • PROTEINS
  • Primates Mammalia Vertebrata Chordata Animalia (Animals, Chordates, Humans, Mammals, Primates, Vertebrates) - Hominidae [86215] human common
  • antigens
  • complementary-printed substrate
  • glass substrates
  • modular nucleic acid
  • peptide-major histocompatibility tetrameric complexes
  • restriction endonuclease 9075-08-5 EC 3.1.21.3
  • single-stranded DNA oligomers
  • 10060, Biochemistry studies - General
  • 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
  • Clinical Immunology
  • Methods and Techniques
  • Oncology
  • cancer Neoplasms (MeSH) neoplastic disease
  • cellular suspension
  • facile assembly
  • homogeneity
  • modular assembly
  • repeatability
  • surface-bound scheme
  • versatility
  • Human Medicine, Medical Sciences
  • antigen-specific T-cells immune system
  • downstream analysis laboratory techniques
  • nucleic acid cell sorting NACS laboratory techniques
  • protein microarray laboratory techniques

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