Transcription factor expression for identifying B cell subsets in cattle by flow cytometry

Area of Science:

• Immunology
• Veterinary Science
• Cell Biology

Background:

• B cells are crucial for immune responses but can cause disease.
• Studying B cell differentiation is vital for understanding immunity and autoimmunity.
• Limited immunological tools hinder B cell subset research in livestock, particularly cattle.

Purpose of the Study:

• To establish novel methods for identifying major cattle B cell subsets using flow cytometry.
• To characterize total mature B cells, plasma cells/plasmablasts, and germinal center B cells in cattle.
• To enable the study of antigen-driven B cell responses in cattle.

Main Methods:

• Utilized cross-reacting anti-human/anti-murine antibodies combined with anti-bovine reagents.
• Employed antibodies targeting CD79α, Pax5, Blimp1, IRF4, and Bcl6 for flow cytometry.
• Used foot-and-mouth disease virus-like particles (VLPs) to detect antigen-specific B cells.

Main Results:

• Successfully identified three major cattle B cell subsets: total mature B cells, plasma cells/plasmablasts, and germinal center B cells.
• Characterized B cell phenotypes based on co-expression of specific markers (e.g., CD79α/Pax5, Blimp1/IRF4, Bcl6).
• Demonstrated the ability to dissect antigen-driven B cell responses using established phenotypes, with antigen-specific cells concentrated in lymph nodes.

Conclusions:

• Developed a significantly expanded immunological toolbox for cattle B cell research.
• Provided new phenotypes for identifying distinct B cell populations in cattle.
• Enabled detailed analysis of B cell differentiation and antigen-specific responses in livestock.