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. Author manuscript; available in PMC: 2013 Nov 20.
Published in final edited form as: J Immunol Methods. 2010 Jan 28;354(0):10.1016/j.jim.2010.01.007. doi: 10.1016/j.jim.2010.01.007

Figure 4. CD B cells rapidly differentiate to AFC and increase CD80/CD86 expression upon TLR stimulation.

Figure 4

CD43CD93lo CD B cells and mature splenic B-cell populations (MF and MZ) were sorted using FACS. Sorted cells were cultured with 5µg/ml LPS for 24hr, 48hr or 72hr. (A) Number of IgM AFC was determined by ELISPOT after LPS stimulation for mature (♦), MZ (▪) or CD (▲) B cells. ELISPOT plates coated with anti-mouse Ig(H+L) polyclonal Ab and assays were performed in triplicate for each condition tested. Values represent mean and standard deviation of triplicate samples. (B) Expression of CD80 and CD86 was measured on mature, MZ and CD43CD93lo CD B cells after LPS stimulation. Cells were labeled with mAbs to B220, CD80 and CD86 after LPS treatment for 48hr (blue) or 72hr (green) and compared to ex vivo splenocytes (red).