Estimation of low frequency antigen-presenting cells with a novel RELISPOT assay
Graduate School of Biomedical Sciences; Vaccine Branch
Medical Subject Headings
Animals; Antigen Presentation; Antigen-Presenting Cells; Antigens, Viral; CD8-Positive T-Lymphocytes; Dendritic Cells; Enzyme-Linked Immunosorbent Assay; Epitopes, T-Lymphocyte; Female; Flow Cytometry; Fluorescent Dyes; Interferon-gamma; Mice; Mice, Inbred BALB C; Microscopy, Fluorescence; Ovary; Rosette Formation; Vaccinia; Vaccinia virus
Life Sciences | Medicine and Health Sciences
Adequate presentation of self and foreign antigens is a key factor for efficient T-cell immunosurveillance against pathogens and tumors. Cells presenting foreign antigens usually comprise a rare population and are difficult to detect even at the peak of infection. Here we demonstrate a CD8(+) T-cell-based approach that allows detection of specific antigen-presenting cells (APC) at a frequency of less than 0.0005%. When T cells are in excess, they form rosettes with rare APCs, which appear as single spots in an IFN-gamma ELISPOT assay. Using this RELISPOT (Rosette ELISPOT) method we demonstrate the dynamic interplay between CD8 T cells and professional and non-professional APCs following virus challenge.
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Citation: J Immunol Methods. 2008 Apr 20;333(1-2):71-8. Epub 2008 Feb 8. Link to article on publisher's site
Dzutsev, Amiran K.; Belyakov, Igor M.; Isakov, Dmitry V.; Gagnon, Susan J.; Margulies, David H.; and Berzofsky, Jay A., "Estimation of low frequency antigen-presenting cells with a novel RELISPOT assay" (2008). GSBS Student Publications. Paper 1458.