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FluoroSpot

About: FluoroSpot is a research topic. Over the lifetime, 64 publications have been published within this topic receiving 1666 citations.


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Journal ArticleDOI
TL;DR: A solid-phase enzyme-linked immunosorbent assay (ELISPOT) is described, which provides a useful alternative to conventional plaque-forming cell assays.

1,047 citations

Journal ArticleDOI
TL;DR: A Fluorospot assay to detect single cells that simultaneously produce multiple cytokines that will be useful for detailed analysis of T lymphocytes in various disease states in which an imbalance of T cell subpopulations is suspected, but will also provide a better characterization of polarized specific immune responses.

98 citations

Journal ArticleDOI
TL;DR: This study detected an expansion of SARS-CoV-2 nucleocapsid protein–specific Ab-secreting cells in all 20 COVID-19 patients using a multicolor FluoroSpot Assay, and showed that IL-6 and C-reactive protein serum concentrations were higher in patients who were hospitalized for longer, supporting the recent observations that IL/C- reactive protein could be used as markers for CO VID-19 severity.
Abstract: COVID-19, caused by SARS-CoV-2, emerged in late 2019 and has since become a global pandemic. Pathogen-specific antibodies are typically a major predictor of protective immunity, yet B cell and antibody responses during COVID-19 are not fully understood. Here, we analyzed antibody-secreting cell (ASC) and antibody responses in twenty hospitalized COVID-19 patients. We observed a significant expansion of SARS-CoV-2 nucleocapsid protein-specific ASCs in all twenty COVID-19 patients using a multicolor FluoroSpot assay. Out of the 20 patients, 16 had developed SARS-CoV-2-neutralizing antibodies by the time of sampling. Additionally, we found that SARS-CoV-2-specific IgA, IgG and IgM antibody levels positively correlated with SARS-CoV-2-neutralizing antibody titers. This study constitutes a detailed description of B cell and antibody responses to SARS-CoV-2 in COVID-19, and provides tools to study immune responses to SARS-CoV-2 infection and vaccination.

70 citations

Journal ArticleDOI
27 Nov 2014-Cells
TL;DR: 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 systems

41 citations

Journal ArticleDOI
TL;DR: In this study, lipoteichoic acid (LTA)‐ and lipopolysaccharide (LPS)‐induced cytokine secretion by monocytes is investigated using the FluoroSpot technique, which measures the number of cytokine‐secreting cells on the single‐cell level and uses fluorescent detection, allowing for the simultaneous analysis of two cytokines from the same population of isolated cells.
Abstract: Monocytes have long been considered a heterogeneous group of cells both in terms of morphology and function. In humans, three distinct subsets have been described based on their differential expression of the cell surface markers CD14 and CD16. However, the relationship between these subsets and the production of cytokines has for the most part been based on ELISA measurements, making it difficult to draw conclusions as to their functional profile on the cellular level. In this study, we have investigated lipoteichoic acid (LTA)- and lipopolysaccharide (LPS)-induced cytokine secretion by monocytes using the FluoroSpot technique. This method measures the number of cytokine-secreting cells on the single-cell level and uses fluorescent detection, allowing for the simultaneous analysis of two cytokines from the same population of isolated cells. By this approach, human monocytes from healthy volunteers could be divided into several subgroups as IL-1β, IL-6, TNF-α and MIP-1β were secreted by larger populations of responding cells (25.9–39.2%) compared with the smaller populations of GM-CSF (9.1%), IL-10 (1.3%) and IL-12p40 (1.2%). Furthermore, when studying co-secretion in FluoroSpot, an intricate relationship between the monocytes secreting IL-1β and/or IL-6 and those secreting TNF-α, MIP-1β, GM-CSF, IL-10 and IL-12p40 was revealed. In this way, dissecting the secretion pattern of the monocytes in response to TLR-2 or TLR-4 stimulation, several subpopulations with distinct cytokine-secreting profiles could be identified.

34 citations

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Performance
Metrics
No. of papers in the topic in previous years
YearPapers
20221
20218
20208
20195
201813
20171