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Decrease in Proportion of CD19⁺CD24^hiCD27⁺ B Cells and Impairment of Their Suppressive Function in Graves’ Disease

Figure 4

CD19⁺CD24^hiCD27⁺ B cells from healthy individuals regulate CD4⁺ T cell proliferation and cytokine production.

(A) Purified CD19⁺CD24^hiCD27⁺(CD24^hiCD27⁺) B cells isolated from blood were cultured with CD40L+CpG, in the presence or absence of blocking antibodies to IL-10. Any IL-10 secreted into the culture supernatant was quantified by ELISA. (B) Purified CD4⁺ cells from the blood of healthy individuals were cultured with media alone or purified CD19⁺CD24^hiCD27⁺ B cells, then stimulated with 0.5 mg/ml of a plate-bound CD3 monoclonal antibody and CD40L+CpG with or without blocking antibodies to IL-10. Proliferation was assessed using Brdu 120 h later. After 120 h, the CD4⁺ cells as in (B) were stained for IFN-γ and TNF-α. Expression of TNF-α and IFN-γ was assessed by flow cytometry. The bar chart shows cumulative data as mean±SEM of three or more independent experiments. Compared to healthy CD24^hiCD27⁺ group, **P<0.01, ***p<0.0001.

doi: https://doi.org/10.1371/journal.pone.0049835.g004