Journal article
Modular Nucleic Acid Assembled p/MHC Microarrays for Multiplexed Sorting of Antigen-Specific T Cells
NanoSystems Biology Cancer Center, Division of Engineering and Applied Science, Bioengineering, Division of Physics, Mathematics and Astronomy, Division of Chemistry and Chemical Engineering, MC 127-72, California Institute of Technology, Pasadena, California 91125; Department of Immunology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands; Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, Department of Medicine, Division of Hematology-Oncology, Department of Surgery, Division of Surgical Oncology, Department of Microbiology, Immunology, and Molecular Genetics, Howard Hughes Medical Institute; University of California, Los Angeles, California 90095
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 offers 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.
Language: | English |
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Publisher: | American Chemical Society |
Year: | 2009 |
Pages: | 9695-9703 |
ISSN: | 15205126 and 00027863 |
Types: | Journal article |
DOI: | 10.1021/ja9006707 |