Sorting antigen-specific T cells

PBMCs from patient KI-705 were stained with both PE-conjugated YF9-tetramer and APC-conjugated FF9-tetramer and then stained with FITC-anti-CD3 (Dako), Pacific Blue-anti-CD8 (BD Pharmingen, USA), and 7-AAD. Among CD3⁺CD8⁺7-AAD⁻ cells, WT-specific, cross-reactive, or MT-specific cells were sorted into a 96-well plate by using a FACS Aria I instrument. Unbiased identification of TCR gene usage was assessed as described previously (51 (link)).

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Kuse N., Murakoshi H., Akahoshi T., Chikata T., James K.L., Gatanaga H., Rowland-Jones S.L., Oka S, & Takiguchi M. (2021). Collaboration of a Detrimental HLA-B*35:01 Allele with HLA-A*24:02 in Coevolution of HIV-1 with T Cells Leading to Poorer Clinical Outcomes. Journal of Virology, 95(23), e01259-21.

Publication 2021

FITC-anti-CD3 Pacific Blue-anti-CD8

7 aad Anti cd3 Aria Cd8 cells Cells Cross reactive Fitc Patient Tcr gene Tetramer

Corresponding Organization : Kumamoto University

Other organizations : University of Oxford, National Center for Global Health and Medicine

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Variable analysis

independent variables

Staining with PE-conjugated YF9-tetramer
Staining with APC-conjugated FF9-tetramer
Staining with FITC-anti-CD3 (Dako)
Staining with Pacific Blue-anti-CD8 (BD Pharmingen, USA)

dependent variables

Sorting of WT-specific, cross-reactive, or MT-specific cells into a 96-well plate

control variables

Staining with 7-AAD to exclude dead cells
Gating on CD3+CD8+ cells

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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