We are interested in determining CD4+ and CD8+ T cell response to recombinant MVA.
To measure anti-vaccinia response, we coculture effector PBMC with autologous APC previously infected with recombinant MVA. After 16h of incubation period, we perform an intracellular staining to study cytokine production.
I’m wondering whether adding CD28/CD49d costimulation would enhance CD4+ and CD8+ T cell response.
Hi Samandhy,
question: what is effector PBMC?
Comment: if you co-culture DC and effector T cell CD4 or CD8 or both I will let them 3 to 4 day to proliferate. After add brefeldinA plus or minus CD3/CD28 5h to 12h. And do the intra-FACS.
best
Hi
Make sure your APC have the high levels of co-stimulatory factors: CD80/83/86. Otherwise, they could induce a TH2-type response. Some APC infected with viruses (like dengue), have high TH1-type response. Others, like measles, can reduce stimulation to blunt the cellular immune response. I think vaccinia is somewhere in the middle, or towards the TH1-type.
Bruce
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