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OMIP-043

Identification of human antibody secreting cell subsets

Jeffrey Carrell, Christopher J. Groves Cytometry PART A, Volume 93, Issue 2, 190–193 (2018)

PURPOSE: This panel was optimized primarily to determine the frequency and immunophenotype of antibody secreting cells (ASC), historically called plasma B cells [PCs, reviewed in Refs. ( 1-3)]. The panel can also be used to determine the frequency and phenotype of other B cell subsets including memory B cells and naïve B cells, which occur in various anatomic niches, but particularly in the circulation (Table 1). The panel has been tested on various human tissues from healthy subjects, and has been shown to be applicable to fresh and cryopreserved peripheral blood, spleen, bone marrow, and tonsil cells; we have observed active antibody secretion from thawed cells. We continue to apply the panel to a variety of tissues and donors to build on our understanding of humoral immunity and the cells that contribute to long‐lasting vaccine responses. Analysis of rare cells can be difficult, and our aim is to encourage better data in the analysis of ASC.

CELL TYPE: Fresh or cryopreserved PBMC, bone marrow, spleen, or tonsil cells

MACHINE: BD SORP Fortessa

Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Blue viability
CD20 Human 2H7 Brilliant Violet 421 B cell differentiation
CD27 Human Brilliant Violet 785 B cell differentiation
CD38 Human HB7 BUV395 Primary identificaiton of ASCs
CD19 Human SJ25C1 APC B cell differentiation
IgD Human IA6-2 PerCP-Cy5.5 B cell maturation
CD3 Human UCHT1 Brilliant Violet 510 Dump/exclusion
CD14 Human MfP9 Brilliant Violet 510 Dump/exclusion
CD15 Human W6D3 Brilliant Violet 510 Dump/exclusion
CD193 Human Brilliant Violet 510 Dump/exclusion
IgM Human G20-127 Brilliant Violet 605 surface Ig
IgM Human G18-145 FITC Cytoplasmic Ig
IgG Human Brilliant Violet 605 surface Ig
IgG Human G20-127 FITC Cytoplasmic Ig
IgA Human PAb FITC Cytoplasmic Ig
IgE Human PE-Cy7 Cytoplasmic Ig
Ki-67 Human N/A PE Proliferation