Detection and Characterizationof Cellular Immune Responses Using Peptide–MHC Microarrays

Soen, Yoav; Chen, Daniel S.; Kraft, Daniel; Davis, Mark M.; Brown, Patrick O.

doi:10.1371/journal.pbio.0000065

Cited by 136 publications

(116 citation statements)

References 31 publications

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“…Fluorescently labeled MHC multimers are commonly used for the detection of antigen-specific T cells by flow cytometry. In addition, there is an increasing interest in the development of high-throughput assay systems, such as MHC microarrays or combinatorial coding schemes, to visualize pathogen-specific or other disease-associated immune responses in a more comprehensive manner (2,3,21). A major obstacle in the development of these high-throughput approaches for the dissection of antigen-specific CD8 ϩ T cell immunity has been the fact that for each specific peptide-MHC class I complex, a separate production run is required (1,22), limiting the practical use of MHC multimer-based T cell detection to a few T cell specificities.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Conditional MHC class I ligands and peptide exchange technology for the human MHC gene products HLA-A1, -A3, -A11, and -B7

Bakker¹,

Hoppes

Linnemann³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

135

167

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Major histocompatibility complex (MHC) class I multimer technology has become an indispensable immunological assay system to dissect antigen-specific cytotoxic CD8 ؉ T cell responses by flow cytometry. However, the development of high-throughput assay systems, in which T cell responses against a multitude of epitopes are analyzed, has been precluded by the fact that for each T cell epitope, a separate in vitro MHC refolding reaction is required. We have recently demonstrated that conditional ligands that disintegrate upon exposure to long-wavelength UV light can be designed for the human MHC molecule HLA-A2. To determine whether this peptide-exchange technology can be developed into a generally applicable approach for high throughput MHC based applications we set out to design conditional ligands for the human MHC gene products HLA-A1, -A3, -A11, and -B7. Here, we describe the development and characterization of conditional ligands for this set of human MHC molecules and apply the peptide-exchange technology to identify melanoma-associated peptides that bind to HLA-A3 with high affinity. The conditional ligand technology developed here will allow high-throughput MHC-based analysis of cytotoxic T cell immunity in the vast majority of Western European individuals.M HC Class I molecules are heterotrimeric complexes consisting of an invariant light chain called ␤2-microglobulin (␤2m), a polymorphic heavy chain (HC) and an Ϸ8-to 11-aa peptide ligand. These peptide-MHC (pMHC) complexes are recognized by the T cell receptor (TCR) of CD8 ϩ T cells in a peptide-specific fashion, and this interaction forms the molecular basis of antigen recognition by CD8 ϩ T cells. In the past decade, the mapping of pathogen-specific and autoimmune-or cancer-associated T cell epitopes has been a major driving force in the development of assay systems for immunomonitoring. In addition, knowledge of such T cell epitopes forms a cornerstone in the development of vaccine-based or adoptive T cell therapies. As a first step in the mapping of disease-associated T cell epitopes, peptide fragments of disease-associated proteomes may be analyzed for binding to MHC molecules of interest, and subsequent assays can then be used to determine whether T cell reactivity against such pMHC complexes does occur. As demonstrated in a landmark study by Altman and colleagues (1), such antigen-specific T cell reactivity can efficiently be detected by the staining of T cell populations with recombinant fluorescent multimeric MHC molecules.There is an increasing interest in the development of assay systems, such as MHC-based microarrays, that can monitor a multitude of T cell responses in parallel (2-4). Unfortunately, current technology does not allow for the high-throughput generation of different pMHC complexes, thereby limiting the utility of these techniques. Specifically, because MHC class I complexes that are devoid of peptide are markedly unstable (5, 6), current production processes for recombinant MHC complexes require inclusion of a specific T cell epi...

show abstract

Section: Discussionmentioning

confidence: 99%

“…There is an increasing interest in the development of assay systems, such as MHC-based microarrays, that can monitor a multitude of T cell responses in parallel (2)(3)(4). Unfortunately, current technology does not allow for the high-throughput generation of different pMHC complexes, thereby limiting the utility of these techniques.…”

mentioning

confidence: 99%

Conditional MHC class I ligands and peptide exchange technology for the human MHC gene products HLA-A1, -A3, -A11, and -B7

Bakker¹,

Hoppes

Linnemann³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

135

167

View full text Add to dashboard Cite

show abstract

“…Peptide-major histocompatibility complex (MHC) tetramer arrays have recently been developed and partially validated for the study of antigen-specific T cells 45 . Individual peptide-MHC tetramer molecules are spotted onto the surface of glass microscope slides at specific positions before being probed with living T cells, which bind to individual tetramer spots and can be quantified and further studied by analysing calcium flux and cytokine secretion.…”

Section: Protein Arrays For Analysing T-cell Receptorsmentioning

confidence: 99%

An array of possibilities for the study of autoimmunity

Fathman

Soares

Chan

et al. 2005

Nature

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Since the completion of the sequencing of the human genome, scientific focus has shifted from studying genes to analysing the much larger number of proteins encoded by them. Several proteins can be generated from a single gene depending on how the genetic information is read (transcribed) and how the resultant protein is modified following translation (post-translational modification). Genomic and proteomic technologies are already providing useful information about autoimmune disease, and they are likely to lead to important discoveries within the next decade.

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“…Variables such as pH, ionic strength, hydration, etc., must be controlled to prevent protein denaturation. The best surface for reducing nonspecific binding of cells while maintaining full antibody functionality is acrylamide (72), which is incompatible with DNA. DNA microarrays are electrostatically absorbed (via spotting) onto amine surfaces.…”

Section: Multiparameter Measurementsmentioning

confidence: 99%

Nanotechnology and Cancer

2004

JAMA

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Detection and Characterizationof Cellular Immune Responses Using Peptide–MHC Microarrays

Cited by 136 publications

References 31 publications

Conditional MHC class I ligands and peptide exchange technology for the human MHC gene products HLA-A1, -A3, -A11, and -B7

Conditional MHC class I ligands and peptide exchange technology for the human MHC gene products HLA-A1, -A3, -A11, and -B7

An array of possibilities for the study of autoimmunity

Nanotechnology and Cancer

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