التفاصيل البيبلوغرافية
| العنوان: |
Vagaries of the ELISpot assay: Specific detection of antigen responsive cells requires purified CD8+ T cells and MHC class I expressing antigen presenting cell lines. |
| المؤلفون: |
Fuchs, Yannick F.1,2,3, Jainta, Gregor W.1, Kühn, Denise1, Wilhelm, Carmen1, Weigelt, Marc1, Karasinsky, Anne1, Upadhyaya, Bhaskar1, Ziegler, Anette-G.3,4, Bonifacio, Ezio1,2,4 ezio.bonifacio@crt-dresden.de |
| المصدر: |
Clinical Immunology. Apr2015, Vol. 157 Issue 2, p216-225. 10p. |
| مصطلحات موضوعية: |
*ENZYME-linked immunosorbent assay, *CD8 antigen, *T cells, *CELL lines, *CYTOKINES, *AUTOANTIGENS |
| مستخلص: |
Quantification of antigen-specific CD8 + T cells is important for monitoring infection, vaccination, and response to therapy in cancer and immune-mediated diseases. Cytokine enzyme-linked-immunospot (ELISpot) assays are often used for this purpose. We found that substantial spot formation in IFNγ ELISpot assays occurred independently of CD8 + T cells even when classical MHC class I restricted peptides are used for stimulation. Using fractionated cells and intracellular cytokine staining, the non-CD8 + T cell IFNγ production was attributed to the CD4 + T cell fraction. We therefore refined a cell line-based ELISpot assay combining HLA-A*0201 expressing K562 cells for antigen presentation with purified CD8 + T cells and demonstrated that it specifically detected CD8 + T cell responses with detection limits comparable to traditional ELISpot assays and dextramer-based quantification. The assay was further adapted to whole antigen responses with antigen (pre-proinsulin)-expressing HLA-A*0201K562 cells. Thus, we revealed and corrected a weak spot of the CD8 + ELISpot assay. [ABSTRACT FROM AUTHOR] |
| قاعدة البيانات: |
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