RhoA is necessary for BAFF-mediated B cell survival but not proliferation.
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Splenic B220+ B cells from control and RhoA mice were cultured for 72 hours on 96-well plates (2×105 cells/well) with or without 2 µg/mL anti-IgM F(ab')2 antibody or indicated concentrations of BAFF (left).
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Identification of IL21, CD40L, αIgM, BAFF and LPS regulated genes in transformed human germinal centre B cells using microarrays.
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BL2 cell were stimulated with αIgM F(ab)2 fragments (3 hrs) , IL21 (2 hrs) , CD40L (6 hrs) , LPS (6 hrs) and BAFF (9 hrs) .
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(A) B cell activating factor (BAFF) mRNA expression in monocytes in patients with primary Sjögren's syndrome (pSS) (n=69) and healthy controls (HC) (n=44).
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BAFF mRNA expression was determined by real time quantitative-PCR.
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(A) B cell activating factor (BAFF) mRNA expression in monocytes in patients with primary Sjögren's syndrome (pSS) (n=69) and healthy controls (HC) (n=44).
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BAFF mRNA expression was determined by real time quantitative-PCR.
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Cell stimulation and total IgG production
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PBMCs recovered from the gradient interface were washed twice in PBS and adjusted to 106 cells/mL in RPMI 1640 supplemented with 10% FCS and 50 μg/mL gentamicin (GIBCO).
- Cells were cultured in the presence of 100 ng/mL rhIL-21 and/or 100 ng/mL rhBAFF for 12 days at 37°C with 5% CO2.
- After 12 days, culture supernatants were collected and total IgG was measured using an in-house ELISA as described previously [
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