Estimation of low frequency antigen-presenting cells with a novel RELISPOT assay

• Published in: Journal of immunological methods Vol. 333; no. 1; pp. 71 - 78
• Main Authors: Dzutsev, Amiran K., Belyakov, Igor M., Isakov, Dmitry V., Gagnon, Susan J., Margulies, David H., Berzofsky, Jay A.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 20.04.2008; Elsevier
• Subjects: Animals; Antigen Presentation; Antigen-presenting cell; Antigen-Presenting Cells - immunology; Antigen-Presenting Cells - virology; Antigens, Viral - immunology; Biological and medical sciences; CD8-Positive T-Lymphocytes - immunology; CD8-Positive T-Lymphocytes - virology; Dendritic cell; Dendritic Cells - immunology; ELISPOT; Enzyme-Linked Immunosorbent Assay - methods; Epitopes, T-Lymphocyte; Female; Flow Cytometry; Fluorescent Dyes - chemistry; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Interferon-gamma - biosynthesis; Interferon-gamma - immunology; Mice; Mice, Inbred BALB C; Microbiology; Microscopy, Fluorescence; Migration; Molecular immunology; Ovary - immunology; Ovary - virology; RELISPOT; Rosette Formation; T-cell line; Techniques; Techniques used in virology; Vaccinia - immunology; Vaccinia virus; Vaccinia virus - immunology; Virology; Vaccinia virus; Dendritic cell; LN; APC; Migration; RELISPOT; Antigen-presenting cell; T-cell line; ELISPOT; DC; Chordopoxvirinae; Orthopoxvirus; Accessory cell; Immunological method; Virus; Cell line; Poxviridae; T-Lymphocyte; ELISPOT assay
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/j.jim.2008.01.008

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-γ 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.