OMIP-043
Identification of human antibody secreting cell subsets
Cytometry PART A, Volume 93, Issue 2, 190–193 (2018)
Jeffrey Carrell, Christopher J. Groves
Description: This panel was optimized primarily to determine the frequency and immunophenotype of antibody secreting cells (ASC), historically called plasma B cells. 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. 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.
Species: Human
Tissue: Bone marrow, Peripheral blood, Spleen, Tonsil
Cell Subset: Plasma cell/ asc, B cell, B memory
Keyword: Vaccination
Machine: BD SORP Fortessa
Panel details
Marker | Species | Clone | Fluorophore | Purpose | Co-expressions groups | Antigen Density |
---|---|---|---|---|---|---|
Dead cells | Human | Live/Dead Fix Blue | viability | |||
CD20 | Human | 2H7 | Brilliant Violet 421 | B cell differentiation | B cell, B memory, B naive | Med |
CD27 | Human | O323 | Brilliant Violet 785 | B cell differentiation | B cell, B memory | Med |
IgD | Human | IA6-2 | PerCP-Cy5.5 | B cell maturation | B cell, B memory, B naive | High |
CD3 | Human | UCHT1 | Brilliant Violet 510 | Dump/exclusion | High | |
CD14 | Human | MfP9 | Brilliant Violet 510 | Dump/exclusion | High | |
IgM | Human | G18-145 | FITC | Cytoplasmic Ig | B Progenitor, Plasma cell | High |
IgG | Human | G20-127 | FITC | Cytoplasmic Ig | B cell, B memory, Plasma cell | High |
IgA | Human | PAb | FITC | Cytoplasmic Ig | B cell, B memory, Plasma cell | High |
Ki-67 | Human | Ki-67 | PE | Proliferation | B cell, B Progenitor, Plasma cell | Low |
CD15 | Human | W6D3 | Brilliant Violet 510 | Dump/exclusion | High | |
CD38 | Human | HB7 | Brilliant Ultraviolet 395 | Primary identificaiton of ASCs | B cell, B naive, Plasma cell | Med |
IgG | Human | G18-145 | Brilliant Violet 605 | surface Ig | B cell, B memory | High |
IgE | Human | MHE-18 | PE-Cy7 | Cytoplasmic Ig | B cell, B memory, Plasma cell | Med |
CD19 | Human | SJ25C1 | APC | B cell differentiation | B cell, B memory, B naive, Plasma cell | Low |
CD193 | Human | 5E8 (also known as 5E8-G9-B4) | Brilliant Violet 510 | Dump/exclusion | Med | |
IgM | Human | G20-127 | Brilliant Violet 605 | surface Ig | B cell, B naive | Med |