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231224s2015 xx |||||o 00| ||eng c |
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|a 10.1016/j.clim.2015.02.012
|2 doi
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|a pubmed24n0822.xml
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|a (DE-627)NLM24667105X
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|a (NLM)25728493
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|a (PII)S1521-6616(15)00061-3
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|a DE-627
|b ger
|c DE-627
|e rakwb
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|a eng
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| 100 |
1 |
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|a Fuchs, Yannick F
|e verfasserin
|4 aut
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1 |
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|a Vagaries of the ELISpot assay
|b specific detection of antigen responsive cells requires purified CD8(+) T cells and MHC class I expressing antigen presenting cell lines
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|c 2015
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
|b c
|2 rdamedia
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|a ƒa Online-Ressource
|b cr
|2 rdacarrier
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|a Date Completed 29.06.2015
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|a Date Revised 27.04.2015
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a Copyright © 2015 Elsevier Inc. All rights reserved.
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|a 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
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|a Journal Article
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|a Research Support, Non-U.S. Gov't
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|a Autoantigen
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| 650 |
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|a CD8(+) T cells
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| 650 |
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|a Diabetes
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|a ELISpot
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|a MHC class I
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| 650 |
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|a HLA-A*02:01 antigen
|2 NLM
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| 650 |
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|a HLA-A2 Antigen
|2 NLM
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| 650 |
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|a Histocompatibility Antigens Class I
|2 NLM
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| 650 |
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|a Interferon-gamma
|2 NLM
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| 650 |
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|a 82115-62-6
|2 NLM
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| 700 |
1 |
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|a Jainta, Gregor W
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Kühn, Denise
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Wilhelm, Carmen
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Weigelt, Marc
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Karasinsky, Anne
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Upadhyaya, Bhaskar
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Ziegler, Anette-G
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Bonifacio, Ezio
|e verfasserin
|4 aut
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| 773 |
0 |
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|i Enthalten in
|t Clinical immunology (Orlando, Fla.)
|d 1999
|g 157(2015), 2 vom: 15. Apr., Seite 216-25
|w (DE-627)NLM098196855
|x 1521-7035
|7 nnns
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| 773 |
1 |
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|g volume:157
|g year:2015
|g number:2
|g day:15
|g month:04
|g pages:216-25
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| 856 |
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|u http://dx.doi.org/10.1016/j.clim.2015.02.012
|3 Volltext
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