Sorting CD8+ T Cell Subsets

CD3⁺CD8^– T lymphocytes were negatively selected from PBMCs and sorted on the BD FACS Aria III sorter into the following subsets: CD95^– naive T cells (CD45RA⁺CCR7⁺CD27⁺CD95^–), Tcm (CD45RA^–CCR7⁺CD27⁺), Ttm (CD45RA^–CCR7^–CD27⁺), and Tem cells (CD45RA^–CCR7^–CD27^–), as described in ref. 10 (link), using BV421 anti-CD45RA (clone 5H9, BD Biosciences), PE-Cy7 anti-CD27 (clone 0323, Invitrogen), APC anti-CD95 (clone DX2, BD Biosciences), and BB700 anti-CCR7 (clone 3D12, BD Biosciences). FlowJo v10.6 software was used for analysis.

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Pinzone M.R., Weissman S., Pasternak A.O., Zurakowski R., Migueles S, & O’Doherty U. (2021). Naive infection predicts reservoir diversity and is a formidable hurdle to HIV eradication. JCI Insight, 6(16), e150794.

Publication 2021

BD FACS Aria III sorter BV421 anti-CD45RA (clone 5H9, PE-Cy7 anti-CD27 (clone 0323, APC anti-CD95 (clone DX2, BB700 anti-CCR7 (clone 3D12,

Aria Cd8 t lymphocytes Cells Clone T cells

Corresponding Organization : University of Pennsylvania

Other organizations : University of Amsterdam, Amsterdam University Medical Centers, University of Delaware, National Institute of Allergy and Infectious Diseases

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Variable analysis

independent variables

CD45RA

dependent variables

T cell subset composition

control variables

Negative selection of CD3+CD8- T lymphocytes from PBMCs
Sorting using BD FACS Aria III sorter

controls

No positive or negative controls were explicitly mentioned in the protocol.

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