The use of fluorescent target arrays for assessment of T cell responses in vivo

Simple item page
dc.contributor.authorQuah, B.
dc.contributor.authorWijesundara, D.
dc.contributor.authorRanasinghe, C.
dc.contributor.authorParish, C.
dc.date.issued2014
dc.descriptionVideo Article. Date Published: 6/19/2014
dc.description.abstractThe ability to monitor T cell responses in vivo is important for the development of our understanding of the immune response and the design of immunotherapies. Here we describe the use of fluorescent target array (FTA) technology, which utilizes vital dyes such as carboxyfluorescein succinimidyl ester (CFSE), violet laser excitable dyes (CellTrace Violet: CTV) and red laser excitable dyes (Cell Proliferation Dye eFluor 670: CPD) to combinatorially label mouse lymphocytes into > 250 discernable fluorescent cell clusters. Cell clusters within these FTAs can be pulsed with major histocompatibility (MHC) class-I and MHC class-II binding peptides and thereby act as target cells for CD8⁺ and CD4⁺ T cells, respectively. These FTA cells remain viable and fully functional, and can therefore be administered into mice to allow assessment of CD8⁺ T cell-mediated killing of FTA target cells and CD4⁺ T cell-meditated help of FTA B cell target cells in real time in vivo by flow cytometry. Since > 250 target cells can be assessed at once, the technique allows the monitoring of T cell responses against several antigen epitopes at several concentrations and in multiple replicates. As such, the technique can measure T cell responses at both a quantitative (e.g. the cumulative magnitude of the response) and a qualitative (e.g. functional avidity and epitope-cross reactivity of the response) level. Herein, we describe how these FTAs are constructed and give an example of how they can be applied to assess T cell responses induced by a recombinant pox virus vaccine.
dc.description.statementofresponsibilityBenjamin J. C. Quah, Danushka K. Wijesundara, Charani Ranasinghe, Christopher R. Parish
dc.identifier.citationJournal of Visualized Experiments, 2014; (88):e51627-1-e51627-10
dc.identifier.doi10.3791/51627
dc.identifier.issn1940-087X
dc.identifier.issn1940-087X
dc.identifier.orcidWijesundara, D. [0000-0002-0740-8362]
dc.identifier.urihttp://hdl.handle.net/2440/113534
dc.language.isoen
dc.publisherJournal of Visualized Experiments
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/1010395
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/525431
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/455395
dc.rightsCopyright © 2014 Journal of Visualized Experiments
dc.source.urihttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4195613/
dc.subjectImmunology; Issue 88; investigative Techniques; T cell response; flow cytometry; multiparameter; CTL assay in vivo; carboxyfluorescein succinimidyl ester (CFSE); CellTrace Violet (CTV); Cell Proliferation Dye eFluor 670 (CPD)
dc.titleThe use of fluorescent target arrays for assessment of T cell responses in vivo
dc.typeJournal article
pubs.publication-statusPublished

Files

Collections

Medicine publications