A flexible MHC class I multimer loading system for large-scale detection of antigen-specific T cells

Luimstra, Jolien J.; Garstka, Malgorzata; Roex, Marthe C.J.; Redeker, Anke; Janssen, George M.C.; Veelen, Peter A. van; Arens, Ramon; Falkenburg, J.H. Frederik; Neefjes, Jacques; Ovaa, Huib

doi:10.1084/jem.20180156

Cited by 40 publications

(41 citation statements)

References 45 publications

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“…This UV‐mediated exchange can be performed in a multi‐well format, allowing the generation of thousands of unique pMHC complexes in parallel. Alternative methods for the high‐throughput refolding of MHC molecules with peptides of choice include the use of temperature‐labile peptides, periodate‐cleavable peptides, azobenzene‐containing peptides that may be cleaved by sodium dithionite, or the use of certain di‐peptides that bind specifically to the F pocket of MHC class I molecules, catalyzing rapid exchange with peptides in the environment .…”

Section: Biological Applicationsmentioning

confidence: 99%

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

et al. 2019

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These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer‐reviewed by leading experts in the field, making this an essential research companion.

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Section: Biological Applicationsmentioning

confidence: 99%

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

et al. 2019

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“…Following UV radiation, the peptide in the groove breaks and can be exchanged by a different peptide in the solution. This technique, and equivalent techniques established later, allows for the production of thousands of different monomeric molecules from one single expression of pMHC molecules and hence, T‐cell responses can be screened against large peptide libraries . As for pMHC class II molecules on the other hand, it has been more challenging to develop efficient peptide exchange assays.…”

Section: High‐throughput Production Of Pmhc Tetramersmentioning

confidence: 99%

Peptide‐MHC class I and class II tetramers: From flow to mass cytometry

Christophersen

2020

HLA

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Funding information Stiftelsen KG JebsenTo develop better vaccines and more targeted treatments for cancer and autoimmune disorders, the disease-specific T cells and their cognate antigens need to be better characterized. For more than two decades, peptide-major histocompatibility complex (pMHC) tetramers and flow cytometry have been the gold standard for detection of CD8+ and CD4+ T cells specific to antigens in the context of MHC class I and class II, respectively. Nonetheless, more recent studies combining such reagents with mass cytometry, that is, cytometry by time of flight (CyTOF), have offered far more comprehensive profiling of antigen-specific T-cell responses. In addition, mass cytometry has enabled ex vivo screening of CD8+ T-cell reactivities against hundreds of MHC class I restricted candidate epitopes. MHC class II molecules, on the other hand, have been challenging to combine with mass cytometry as they are more complex and bind with lower affinities to cognate T-cell receptors than MHC class I molecules. In this review, I discuss how techniques originally developed to improve the staining capacity of pMHC tetramers in flow cytometry led to the successful combination of such reagents with mass cytometry. Especially, I will highlight very recent advances facilitating the combination with pMHC class II tetramers. Together, these mass cytometry-based studies can help develop more targeted treatments for cancer and autoimmune disorders.

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“…This elegant technical advance has provided an enormous boost to prepare MHC-I-peptide tools for the detection of circulating T-lymphocytes of a given antigen specificity, extremely relevant to study immune responses to cancer and infection. More recently, he and his colleagues expanded on this early work and developed an efficient method to generate many different MHC-I multimers in parallel using temperature-mediated peptide exchange (Luimstra et al, 2018). Aysegul Sapmaz says: "I came to Huib's lab as a visiting Ph.D. student with a hardcore molecular biology background but a novice in chemistry.…”

Section: Major Contributions To the Ubiquitin And Chemical Biology Fieldmentioning

confidence: 99%

In Memoriam: Professor Huib Ovaa (1973-2020): A Uniquely Brilliant and Enthusiastic Scientist, a Pioneer in Chemical Biology and the Ubiquitin Field

Mulder

Kessler

2020

Front. Chem.

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A flexible MHC class I multimer loading system for large-scale detection of antigen-specific T cells

Abstract: Luimstra et al. describe a temperature-mediated peptide exchange method for generating many different epitope-specific MHC class I multimers in parallel. This simple and versatile technology allows fast and efficient production of MHC I reagents for immune monitoring of T cell responses.

Cited by 40 publications

References 45 publications

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

Peptide‐MHC class I and class II tetramers: From flow to mass cytometry

In Memoriam: Professor Huib Ovaa (1973-2020): A Uniquely Brilliant and Enthusiastic Scientist, a Pioneer in Chemical Biology and the Ubiquitin Field

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