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OMIP-047

High‐Dimensional phenotypic characterization of B cells

Thomas Liechti, Huldrych F. Günthard, Alexandra Trkola Cytometry PART A, Volume 93, Issue 6, 592-596 (2018)

PURPOSE: This 16‐color, 18‐parameter panel was designed to allow a detailed dissection of human B cell subsets and their phenotype in peripheral blood mononuclear cells (PBMC) in healthy donors and in the context of chronic viral diseases such as Human Immunodeficiency Virus 1 (HIV‐1) infection. The panel encompasses a range of backbone markers for the accurate definition of common B cell subsets with a focus on memory B cells and a unique collection of phenotypic markers (chemokine receptors, cytokine receptor, B cell receptor isotypes, and proliferation marker) not combined in multicolor flow cytometry B cell phenotyping thus far. This new panel allows highly detailed phenotypic and functional investigations of B cell subsets. The panel was validated using cryopreserved PBMC from healthy and HIV‐1 infected donors allowing the retrospective analysis of clinical samples (Table 1).

CELL TYPE: PBMC

MACHINE: BD Fortessa

Marker Target Clone Fluorochrome Purpose
CD3 Human SK7 APC-Cy7 Dump
CD14 Human hCD14 APC-Cy7 Dump
CD16 Human 3G8 APC-Cy7 Dump
Dead cells Human Live/Dead Fix Near IR (775) Viability
CD19 Human SJ25C1 Brilliant Violet 786 Lineage
CD10 Human HI10a Brilliant Violet 650 B cell subset
CD38 Human HIT2 Alexa Fluor 700 B cell subset
CD21 Human B-ly4 Brilliant Violet 711 B cell subset/Exhaustion
CD27 Human M-T271 PE-CF594 Differentitation, memory
IgG1 Human HP6001 PE IgG1 calss-switching B cell
IgG3 Human FITC IgG3 calss-switching B cell
IgA Human APC IgA calss-switching B cell
ccr7 Human G043H7 Brilliant Violet 605 Migration pattern
CD183 Human G025H7 PE-Cy7 Migration pattern
CD185 Human RF8B2 Brilliant Violet 510 Migration pattern
IL-21 R Human 2G1-K12 Brilliant Violet 421 Cytokine receptor
Ki-67 Human 20Raj1 PerCP-eFluor 710 Proliferation marker
CXCR4 Human 12G5 PE-Cy5.5 Migration pattern
IgD Human IA6-2 PE-Cy5 B cell subset