using flow for B cell/plasma cell evaluation

Jason, Janine (jmj1@CIDDAS1.EM.CDC.GOV)
Mon, 01 Apr 96 09:13:00 EST

Does anyone have experience using flow to evaluate B cell/plasma cell
populations? Especially:

-Has anyone used flow to evaluate B cell responses to specific antigens?
Can it be done or is this below the sensitivity of the technique? What
antigens are used to identify the B cells or plasma cells?

- As a first step to attempting the above, I have tried looking at human B
cells and plasma cells post "pan-B/plasma cell stimulation" with in vitro
PWM stimulation and find
- the % of cells that are B or plasma is surprisingly low (approx. 10-15%
of lymphocytes)
- I get surface staining with anti-Ig monoclonal antibodies, as well as
cytoplasmic. In fact, my % positive with surface staining is comparable to
the % I get with cytoplasmic staining. Do others see this or is this an
artifact (Fc receptor or something else?)

Many thanks,

Janine Jason
Immunology Branch,
DASTLR, NCID,CDC


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