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ARS Home » Pacific West Area » Pullman, Washington » Animal Disease Research » Research » Publications at this Location » Publication #388962

Research Project: Control Strategies for Bovine Babesiosis

Location: Animal Disease Research

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

item ROOS, EDUARD - The Pirbright Institute
item BONNET-DI PLACIDO, MARIE - The Pirbright Institute
item MWANGI, WILLIAM - The Pirbright Institute
item MOFFAT, KATY - The Pirbright Institute
item Fry, Lindsay
item WATERS, RYAN - The Pirbright Institute
item HAMMOND, JOHN - The Pirbright Institute

Submitted to: Cytometry Journal
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/25/2022
Publication Date: 8/4/2022
Citation: Roos, E.O., Bonnet-Di Placido, M., Mwangi, W., Moffat, K., Fry, L.M., Waters, R., Hammond, J.A. 2022. OMIP-085: Cattle B-cell phenotyping by an 8-color panel. Cytometry Journal. 103(1):12-15.

Interpretive Summary: Study of the immune response in livestock is often greatly limited by the lack of availability of reagents and assays to study various white blood cell types in these species. One specific area in which reagents are limited is the study of bovine B cells and plasma cells, the cells responsible for antibody production during infection. The goal of this study was to develop and optimize a flow cytometry-based panel that utilizes monoclonal antibodies to eight different cell surface proteins to reliably identify different bovine B cell subsets. Using this panel, we were able to label naive B cells (B cells that have not yet participated in immune response development), regulatory B cells (B cells that help control the immune response), memory B cells (long-lived B cells that provide long-term protection from disease after initial exposure to an antigen), plasmablasts (immature antibody-producing cells), and plasma cells (mature antibody producing cells). This work will facilitate future studies on the bovine B cell response to myriad infectious diseases of importance to livestock producers across the globe.

Technical Abstract: Due to the limited availability of reagents to study immunology in the veterinary field, optimised panels to study B cell phenotypes in livestock need to be established. Therefore, a 10-colour, 12-parameter flow cytometry panel was developed to identify bovine B cells. Fresh and cryopreserved peripheral blood mononuclear cells (PBMCs) were subjected to this panel, which focused on distinguishing B cells into five potential subsets: 1) Naïve B cells (BNaïve), 2) Regulatory B cells (BReg), 3) Memory B cells (BMem), 4) Plasmablasts (PB) and 5) Plasma cells (PC). Monoclonal antibodies against the cell surface immunoglobulin light chain (S-Ig(L)), CD20, CD21, CD40, CD71 and CD138 enabled discriminating 24 unique populations within the CD14-CD40+ cells, including the 5 populations of interest above mentioned. We show that the use of CD3 and CD8a can be included as a further Dump channel to CD14 but does not add a significant gain to the panel’s ability to separate the “Classical” B cells. The panel will allow (i) better characterisation of BNaïve, BMem, BReg, and antibody secreting cells (ASC; PBs and PCs), (ii) isolation of these subsets, and (iii) investigating previously unknown B cell subsets in bovids.