Ly6C expression differentiates plasma cells from other B cell subsets in mice
Jens Wrammert1, Eva Källberg, William W Agace
1Section for Immunology, Department of Cell and Molecular Biology, Lund University, Lund, Sweden. jens.wrammert@immuno.lu.se
Insights
The cell surface protein Ly6C is newly identified on B cells stimulated with lipopolysaccharide (LPS). Ly6C expression on B cells increases during plasma cell differentiation and enhances immunoglobulin production.
Area of Science:
- Immunology
- Cell Biology
Background:
- Plasma cell differentiation is crucial for adaptive immunity.
- Lipopolysaccharide (LPS) and anti-CD40 antibodies are key inducers of B cell activation and plasma cell differentiation.
- The specific markers of B cell activation and plasma cell differentiation are not fully elucidated.
Purpose of the Study:
- To identify novel cell surface markers associated with B cell activation and plasma cell differentiation.
- To investigate the role of the Ly6C protein in B cell responses to LPS stimulation.
- To determine the expression pattern of Ly6C on plasma cells in vivo and in vitro.
Main Methods:
- Subtractive cDNA hybridization was employed to identify differentially expressed genes.
- Flow cytometry and magnetic-activated cell sorting (MACS) were used for cell analysis and sorting.
- ELISPOT assays were performed to quantify antibody-secreting cells.
- In vitro cell culture systems were utilized to study B cell activation and differentiation.
Main Results:
- The cell surface protein Ly6C was identified as differentially expressed on B cells stimulated with LPS but not with anti-CD40 antibodies.
- Ly6C expression was found to be up-regulated on B cells following LPS stimulation in vitro, an effect blocked by anti-CD40 or anti-Ig antibodies.
- Ly6C was confirmed to be expressed on ex vivo plasma cells from spleen and bone marrow, as well as on splenic and lamina propria plasma cells.
- Cross-linking of Ly6C on LPS-stimulated splenic B cells in vitro led to increased immunoglobulin production.
Conclusions:
- Ly6C is a novel marker for activated B cells and plasma cells.
- Ly6C expression is regulated by LPS and immunoglobulin signaling pathways during B cell activation.
- Ly6C plays a functional role in enhancing immunoglobulin production by plasma cells.
Abstract:
Plasma cell differentiation is induced in vitro by lipopolysaccharide (LPS) stimulation but can be blocked by including anti-CD40 antibodies. Using subtractive cDNA hybridization we have identified the cell surface protein Ly6C as differentially expressed on B cells stimulated with LPS only. Ly6C has been shown to be expressed on certain T cell subsets and on subsets of macrophages and NK cells, but not on resting B cells. We show that Ly6C is up-regulated upon LPS stimulation of B cells in vitro and that this up-regulation is blocked by anti-CD40 or anti-Ig antibodies. Furthermore, ELISPOT analysis of cells sorted by magnetic-activated cell sorting show that Ly6C is expressed on ex vivo plasma cells from the spleen and bone marrow. Flow cytometric analysis showed that Ly6C is expressed on splenic plasma cells as well as on lamina propria plasma cells. Finally, Ly6C cross-linking positively up-regulated the amount of immunoglobulin produced by LPS-stimulated splenic B cells in vitro.


