I want to create a panel for T cell and B cell flow cytometry, to investigate how infection affects activation of these cells.
For T cells, CD44 is the classical marker of activation because it remains expressed almost indefinitely once the cell is activated. Know that CD69 and CD25 can be used, but depending on our system, these molecules will have specific kinetics and should eventually come back down. For example, CD69 is a good marker of early activation, being upregulated rapidly after TCR stimulation, but it goes down very quickly once TCR stimulation is removed. As for CD25, in infection models, it is upregulated in the early days of the response, but tends to go back down with decreasing IL-2 availability (see Kalia et al, Immunity, 2010, Shin et al, Immunity, 2013).
Awsome, Thanks,
I believe these markers include adhesion, activation and migration for B cells?.
As for T cells. CD25, CD 28, CD 70, are they sufficient?
For T cells, CD44 is the classical marker of activation because it remains expressed almost indefinitely once the cell is activated. Know that CD69 and CD25 can be used, but depending on our system, these molecules will have specific kinetics and should eventually come back down. For example, CD69 is a good marker of early activation, being upregulated rapidly after TCR stimulation, but it goes down very quickly once TCR stimulation is removed. As for CD25, in infection models, it is upregulated in the early days of the response, but tends to go back down with decreasing IL-2 availability (see Kalia et al, Immunity, 2010, Shin et al, Immunity, 2013).
Depending on the timeframe you are looking at, CD69 is also a good early activation marker for T cells.
For T cells you can use CD44, CD25 and as early activation marker CD69.
For B cells you can use CD86 and MHC-II
CD69 is a good marker for early T cell activation, it is detectable after 4 hrs and peaks at 18-48 hours after stimulation with anti-CD3 (in humans). Ref: PubMedID 8705673.
In mice, murine CD22 is a good B cell activation marker.
If you are measuring T cell activation after in vitro stimulation I would also consider intracellular cytokine staining.
All good answers - I would add one more very early activation marker - CD98. It is an amino acid transporter, broadly expressed. And I should stress once more the kinetics. One can follow the different stages of activation using different kinetics markers, e.g.- early CD69, CD98; intermediate CD25; late HLA-DR; very late - integrin chains, etc.
- Badges
- Science method