Stepping up ELISpot: Multi-Level Analysis in FluoroSpot Assays.
TLDR
The technical and methodological requirements for a reliable analysis of FluoroSpot assays are dissected, important quality control measures are introduced and advice is provided for proper interpretation of results obtained by automated imaging systems.Abstract:
ELISpot is one of the most commonly used immune monitoring assays, which allows the functional assessment of the immune system at the single cell level With its outstanding sensitivity and ease of performance, the assay has recently advanced from the mere single function cell analysis to multifunctional analysis by implementing detection reagents that are labeled with fluorophores (FluoroSpot), allowing the detection of secretion patterns of two or more analytes in a single well However, the automated evaluation of such assays presents various challenges for image analysis Here we dissect the technical and methodological requirements for a reliable analysis of FluoroSpot assays, introduce important quality control measures and provide advice for proper interpretation of results obtained by automated imaging systemsread more
Citations
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Mattias Bronge,Sabrina Ruhrmann,Claudia Carvalho-Queiroz,Ola Nilsson,Andreas Kaiser,Erik Holmgren,Caterina Macrini,Stephan Winklmeier,Edgar Meinl,Lou Brundin,Mohsen Khademi,Tomas Olsson,Guro Gafvelin,Hans Grönlund +13 more
TL;DR: A sensitive method to detect circulating autoreactive CD4+ T-cells producing IFNγ, IL-22 or IL-17A using MOG as a model antigen is presented and it is demonstrated that MOG-specific T- cells are present in approximately half of persons with MS.
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References
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Journal ArticleDOI
A solid-phase enzyme-linked immunospot (ELISPOT) assay for enumeration of specific antibody-secreting cells
TL;DR: A solid-phase enzyme-linked immunosorbent assay (ELISPOT) is described, which provides a useful alternative to conventional plaque-forming cell assays.
Journal ArticleDOI
Improved Endpoints for Cancer Immunotherapy Trials
Axel Hoos,Alexander M.M. Eggermont,Sylvia Janetzki,F. Stephen Hodi,Ramy Ibrahim,Aparna Anderson,R. Humphrey,Brent A. Blumenstein,Lloyd Old,Jedd D. Wolchok +9 more
TL;DR: A body of clinical and laboratory data was generated that supports three novel endpoint recommendations that may improve tools for cancer immunotherapy trials and may offer a more realistic and useful model for clinical investigation.
Journal ArticleDOI
Sensitivity of a new commercial enzyme-linked immunospot assay (T SPOT-TB) for diagnosis of tuberculosis in clinical practice.
TL;DR: The T SPOT-TB test is a sensitive assay for detection of TB and represents a useful addition to the diagnostic algorithm available for detecting TB in low-incidence settings.
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Results and harmonization guidelines from two large-scale international Elispot proficiency panels conducted by the Cancer Vaccine Consortium (CVC/SVI)
Sylvia Janetzki,Katherine S. Panageas,Leah Ben-Porat,Jean D. Boyer,Cedrik M. Britten,Timothy M. Clay,Michael Kalos,Holden T. Maecker,Pedro Romero,Jianda Yuan,W. Martin Kast,Axel Hoos +11 more
TL;DR: These results provide initial harmonization guidelines to optimize Elispot assay performance to the immunotherapy community and are in process with ongoing panels.
Journal ArticleDOI
The CIMT-monitoring panel: a two-step approach to harmonize the enumeration of antigen-specific CD8+ T lymphocytes by structural and functional assays.
Cedrik M. Britten,Cécile Gouttefangeas,Marij J. P. Welters,Graham Pawelec,Sven D. Koch,Christian H. Ottensmeier,Ann Mander,Steffen Walter,Annette Paschen,J. Müller-Berghaus,I. Haas,Andreas Mackensen,Tania Køllgaard,P thor Straten,Michael Schmitt,Krzysztof Giannopoulos,R. Maier,Hendrik Veelken,Cristina Bertinetti,A. Konur,Christoph Huber,Stefan Stevanovic,Thomas Wölfel,S.H. van der Burg +23 more
TL;DR: It is concluded that a two-step approach in inter-laboratory testing allows the identification of distinct variables that influence the sensitivity of different T-cell assays and to formally show that a defined correction to the protocols successfully increases the sensitivity and reduces the inter-center variability.