High Reproducibility of ELISPOT Counts from Nine Different Laboratories
نویسندگان
چکیده
منابع مشابه
High Reproducibility of ELISPOT Counts from Nine Different Laboratories
The primary goal of immune monitoring with ELISPOT is to measure the number of T cells, specific for any antigen, accurately and reproducibly between different laboratories. In ELISPOT assays, antigen-specific T cells secrete cytokines, forming spots of different sizes on a membrane with variable background intensities. Due to the subjective nature of judging maximal and minimal spot sizes, dif...
متن کاملUser vs. software-dependent variability of ELISPOT counts obtained from ten different laboratories
Introduction In each human donor’s PBMC, there is a defined number of T cells specific for any given antigen. A major goal of immune monitoring with ELISPOT is to measure this number accurately and reproducibly between different laboratories. In ELISPOT assays, cytokine spots produced by antigen-specific T cells show a broad spectrum of sizes and densities over variable background. Therefore, e...
متن کاملELISPOT assays provide reproducible results among different laboratories for T-cell immune monitoring--even in hands of ELISPOT-inexperienced investigators.
Measurements of antibodies in bodily fluids (e.g., by ELISA) have provided robust and reproducible results for decades and such assays have been validated for monitoring of B-cell immunity. In contrast, measuring T-cell immunity has proven to be a challenge due to the need to test live cells in functional assays ex vivo. Several previous efforts looking into the reproducibility of ex vivo T-cel...
متن کاملLog-Normal ELISPOT spot size distribution permits count harmonization among different laboratories
Introduction ELISPOT assays are primarily used to detect the number of T cells that respond to a given antigen. With that number being absolute for any given donor sample, ELISPOT counts should be similar between laboratories, if subjectivity in counting is avoided. Due to the differences in spot sizes ranging from microns to millimeters, setting cut offs for minimal and maximal spot sizes will...
متن کاملEquivalence of ELISpot Assays Demonstrated between Major HIV Network Laboratories
BACKGROUND The Comprehensive T Cell Vaccine Immune Monitoring Consortium (CTC-VIMC) was created to provide standardized immunogenicity monitoring services for HIV vaccine trials. The ex vivo interferon-gamma (IFN-γ) ELISpot is used extensively as a primary immunogenicity assay to assess T cell-based vaccine candidates in trials for infectious diseases and cancer. Two independent, GCLP-accredite...
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ژورنال
عنوان ژورنال: Cells
سال: 2015
ISSN: 2073-4409
DOI: 10.3390/cells4010021