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