Design and use of conditional MHC class I ligands

Toebes, Mireille; Coccoris, Miriam; Bins, Adriaan D.; Rodenko, Boris; Gomez, Raquel; Nieuwkoop, Nella J.; Kasteele, Willeke van de; Rimmelzwaan, Guus F.; Haanen, John B.A.G.; Ovaa, Huib; Schumacher, Ton N.

doi:10.1038/nm1360

Cited by 307 publications

(391 citation statements)

References 30 publications

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“…As a step toward this goal, we recently designed an HLA-A2-specific peptide that contains a photocleavable moiety (8). When refolding reactions of HLA-A2 heavy chain and ␤2m are performed with this ligand, stable HLA-A2 complexes are formed.…”

mentioning

confidence: 99%

“…In this case, the peptide-binding groove that has been vacated by UV exposure will be occupied by the rescue peptide, resulting in the formation of stable pMHC complexes with a distinct T cell specificity. The utility of this approach has been demonstrated by the identification of an HLA-A2-restricted CTL epitope from an H5N1 influenza strain isolated from a lethal case of avian influenza infection in humans (8).…”

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confidence: 99%

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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.

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141

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...

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confidence: 99%

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.

Self Cite

141

167

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“…Although MHC class II molecules are traditionally more difficult to develop than class I molecules (49), the peptideexchange method allows for the production of many more tetramer specificities with just a single preparation of refolded monomer (23), and the protocol for tetramer enrichment allows for reliable detection of low-frequency autoreactive T cells (24,25). Future studies are needed to determine whether other autoantigen-specific T cells produce IL-17 or whether this phenotype is unique to U1-70-specific T cells.…”

Section: Discussionmentioning

confidence: 99%

“…We generated recombinant I-E k monomers with a cleavable peptide that could be exchanged out of the I-E k binding cleft for another peptide at low pH (23). Both U1-70 and P140 formed stable complexes with I-E k , similar to MCC (Fig.…”

Section: Significancementioning

confidence: 95%

Tetramers reveal IL-17–secreting CD4⁺T cells that are specific for U1-70 in lupus and mixed connective tissue disease

Kattah

Newell

Jarrell

et al. 2015

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

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Antigen-specific CD4 + T cells are implicated in the autoimmune disease systemic lupus erythematosus (SLE), but little is known about the peptide antigens that they recognize and their precise function in disease. We generated a series of MHC class II tetramers of I-E k -containing peptides from the spliceosomal protein U1-70 that specifically stain distinct CD4 + T-cell populations in MRL/lpr mice. The T-cell populations recognize an epitope differing only by the presence or absence of a single phosphate residue at position serine 140 . The frequency of CD4 + T cells specific for U1-70(131-150):I-E k (without phosphorylation) correlates with disease severity and anti-U1-70 autoantibody production. These T cells also express RORγt and produce IL-17A. Furthermore, the U1-70-specific CD4 + T cells that produce IL-17A are detected in a subset of patients with SLE and are significantly increased in patients with mixed connective tissue disease. These studies provide tools for studying antigen-specific CD4 + T cells in lupus, and demonstrate an antigen-specific source of IL-17A in autoimmune disease.tetramer | autoimmunity | lupus | SLE | IL-17

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“…Although caging of bioactive molecules for subsequent activation by using UV light is a common strategy, [6][7][8][9][10][11] the use of photocleavable functionality for deactivation of peptides and proteins [12] is relatively unexplored. Backbone cleavage through 2-nitrophenyl amino acid derivatives provides an interesting way to specifically turn off a biological signal of interest.…”

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confidence: 99%

Control of the Yeast Cell Cycle with a Photocleavable α‐Factor Analogue

et al. 2006

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The budding yeast Saccharomyces cerevisiae is widely used as a model organism for studying many biological processes and systems, particularly for understanding the cell cycle. The yeast peptide pheromone α-factor (WHWLQLKPGQPMY) activates the mating pathway in MATa cells, arresting the cell cycle in the G1 phase. This arrest is used as a tool for synchronizing yeast cultures and for investigating signaling events involved in the pathway and other processes related to morphogenesis and transcription. Herein we report the synthesis and activity of an α-factor analogue designed to control the on/off state of the mating signal. Incorporating a photocleavable residue at a flexible site in the peptide allowed UV degradation in situ to mimic the natural protease degradation of this signaling peptide. The control over the α-factor signal achieved serves as proof of concept for use of this analogue as a tool. It also and allows access to previously impractical experiments that will yield highly time-resolved information about the yeast cell cycle and mating pathway.

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Design and use of conditional MHC class I ligands

Cited by 307 publications

References 30 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

Tetramers reveal IL-17–secreting CD4⁺T cells that are specific for U1-70 in lupus and mixed connective tissue disease

Control of the Yeast Cell Cycle with a Photocleavable α‐Factor Analogue

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Design and use of conditional MHC class I ligands

Cited by 307 publications

References 30 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

Tetramers reveal IL-17–secreting CD4+T cells that are specific for U1-70 in lupus and mixed connective tissue disease

Control of the Yeast Cell Cycle with a Photocleavable α‐Factor Analogue

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Tetramers reveal IL-17–secreting CD4⁺T cells that are specific for U1-70 in lupus and mixed connective tissue disease