Estimation of low frequency antigen-presenting cells with a novel RELISPOT assay.
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.
Dzutsev
Amiran K
AK
Vaccine Branch, CCR, NCI, NIH, Bethesda, MD 20892-1578, USA.
Belyakov
Igor M
IM
Isakov
Dmitry V
DV
Gagnon
Susan J
SJ
Margulies
David H
DH
Berzofsky
Jay A
JA
D000818:Animals; D017951:Antigen Presentation; D000938:Antigen-Presenting Cells / Q000276:immunology* / Q000821:virology; D000956:Antigens, Viral / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000821:virology; D003713:Dendritic Cells / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D018984:Epitopes, T-Lymphocyte; D005260:Female; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry; D007371:Interferon-gamma / Q000096:biosynthesis / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D010053:Ovary / Q000276:immunology / Q000821:virology; D012397:Rosette Formation; D014615:Vaccinia / Q000276:immunology; D014616:Vaccinia virus / Q000276:immunology*
D000956:Antigens, Viral; D018984:Epitopes, T-Lymphocyte; D005456:Fluorescent Dyes; D007371:Interferon-gamma