OMIP-068: High-dimensional characterization
In this OMIP (Optimised Multicolour Immunophenotyping Panel), the authors describe a 24-color panel to characterise antigen-specific B cells and for the precise delineation of B cell subsets in chronic infection.1 B cells play an important role in infection response through the production of antibodies, antigen presentation and secretion of cytokines. The antibodies produced by B cells against the hepatitis B virus (HBV) envelope protein or the hepatitis B surface antigen (HBsAg), play a key role in the infection response to the virus. The induction of anti-HBsAg is considered as the clinical indication of recovery from HBV. While B cells are an important element of infection response, it is unclear why the response fails in some cases of acute HBV infection. Therefore the authors developed this panel to understand and characterise global and antigen-specific B cells.
Sample extraction and flow cytometry
The authors extracted the cell samples from cryopreserved human PBMC. B cells are characterised by their expression of CD19 and CD20. They used a dump channel to enable the exclusion of CD3+ T cells, CD14+ monocytes and dead cells to improve the resolution of CD19-CD20- and CD19+CD20+ cells. The authors characterised the B cells based on the expression of markers having a specific purpose. These are
- HBV-specific B cell tag: HBsAg
- Lineage B cell markers: CD19, CD20
- Differentiation B cell markers: CD10, CD27, CD21, IgM, IgD, CD24, CD38, CD5, CD43, CD86
- Trafficking markers: CXCR3, CXCR5
- Inhibitory/exhaustion markers: CD11c, CD39, PD-1, FcRL5, BTLA, CD22, CD32
- Markers in the dump channel: CD3, CD14
The authors propose that the panel enables B cell analysis at an unprecedented depth and can potentially uncover new B cell subtypes and phenotypes which could play a role in the progression of infections.
Learn more about the OMIP here.