A new staining protocol for detection of murine antibody-secreting plasma cell subsets by flow cytometry

Area of Science:

• Immunology
• Cell Biology
• Hematology

Background:

• Plasma cells are terminally differentiated B lymphocytes responsible for antibody production.
• Distinguishing between actively dividing plasmablasts and quiescent plasma cells is crucial for understanding humoral immunity.
• Current methods may lack the resolution to differentiate these cell populations effectively.

Purpose of the Study:

• To develop a robust four-color fluorescence-based flow cytometry protocol.
• To accurately distinguish viable dividing plasmablasts from nondividing plasma cells.
• To identify distinct mature plasma cell populations based on CD19 surface expression.

Main Methods:

• Utilized a four-color fluorescence-based flow cytometry approach.
• Applied the protocol to analyze cells from mouse spleen and bone marrow.
• Quantified cell populations based on viability markers and CD19 surface abundance.

Main Results:

• Successfully differentiated viable dividing plasmablasts from nondividing plasma cells.
• Identified two distinct mature plasma cell populations in both spleen and bone marrow.
• Demonstrated the utility of CD19 surface abundance as a marker for plasma cell subsetting.

Conclusions:

• The developed flow cytometry protocol provides a reliable method for dissecting plasma cell heterogeneity.
• This technique facilitates deeper investigation into B cell differentiation and humoral immune responses.
• The identification of distinct plasma cell populations offers new avenues for immunological research.