Dear Christopher,
I added two papers about IL-10 and Bregs. These studies published by my famous friends. They will answer your question.
Dear Christopher D Rudulier
I am not sure, if culturing B cells (derived from PBMCs) with IL10 is the best way to induce a B regulatory cell (like) phenotype. In general, certain bacterial CPGs in combination with CD40L or GMCSF seems to induce IL10+ B cells reliably, as per:
Article Generation of Human Breg-Like Phenotype with Regulatory Func...
In this paper, you also find a simple protocol to assess their suppressive activity and some basic phenotypic characterization.
Similarly you can use variations of this protocol to assess differences in healthy donors and patients with autoimmune diseases (like RA); the complete details you find in the following paper.
Article Induction and Differentiation of IL-10-Producing Regulatory ...
However, it seems these cells are phenotypically heterogeneous as they coexpress both pro- and anti-inflammatory cytokines.
https://www.frontiersin.org/articles/10.3389/fimmu.2018.01913/full
If you plan to look for a simple readout that induces up 15-20% IL10 expression in your B cells, I would suggest to some combination of activation markers that include bacterial CPG (the sequences are buried in the M&M section of the papers) and subsequently set up a co-culture system looking at suppression of T cell proliferation. I hope that helps with the design of your experiment.
All the best & kind regards,
Michael
Hi,
I think you need to go through some of the research articles of the scientists mentioned here, Thomas F. Tedder & Claudia Mauri. They are the pioneers of Breg cells and you can found some amazing concepts and great answers to your questions.
Like, doi: 10.1111/j.1749-6632.2009.05137.x.
Thanks,
Hamid