OMIP‐085: Cattle B‐cell phenotyping by an 8‐color panel

This 8‐color panel has been optimized to distinguish between functionally distinct subsets of cattle B cells in both fresh and cryopreserved peripheral blood mononuclear cells (PBMCs). Existing characterized antibodies against cell surface molecules (immunoglobulin light chain (S‐Ig[L]), CD20, CD21,...

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Bibliographic Details
Published in:Cytometry. Part A 2023-01, Vol.103 (1), p.12-15
Main Authors: Roos, Eduard O., Bonnet‐Di Placido, Marie, Mwangi, William N., Moffat, Katy, Fry, Lindsay M., Waters, Ryan, Hammond, John A.
Format: Article
Language:English
Subjects:
Animals
Antibodies
antibody secreting cells
B cells
B-Lymphocytes, Regulatory
B‐cell subsets
Cattle
CD20 antigen
CD3 antigen
CD40 antigen
CD40 Antigens
Cell surface
Color
Cryopreservation
Flow Cytometry
Immunoglobulins
Immunological memory
Leukocytes (mononuclear)
Lymphocytes B
memory B cells
Memory cells
naïve B cells
Omip
Peripheral blood mononuclear cells
Phenotyping
Plasma cells
Reagents
regulatory B cells
Vaccination
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Summary:This 8‐color panel has been optimized to distinguish between functionally distinct subsets of cattle B cells in both fresh and cryopreserved peripheral blood mononuclear cells (PBMCs). Existing characterized antibodies against cell surface molecules (immunoglobulin light chain (S‐Ig[L]), CD20, CD21, CD40, CD71, and CD138) enabled the discrimination of 24 unique populations within the B‐cell population. This allows the identification of five putative functionally distinct B‐cell subsets critical to infection and vaccination responses: (1) naïve B cells (BNaïve), (2) regulatory B cells (BReg), (3) memory B cells (BMem), (4) plasmablasts (PB), and (5) plasma cells (PC). Although CD3 and CD8α can be included as an additional dump channel, it does not significantly improve the panel's ability to separate “classical” B cells. This panel will promote better characterization and tracking of B‐cell responses in cattle as well as other bovid species as the reagents are likely to cross react.
ISSN:1552-4922
1552-4930
DOI:10.1002/cyto.a.24683