3b) Thus, CD27+ B cells from CVID MB0 patients appear to be resi

3b). Thus, CD27+ B cells from CVID MB0 patients appear to be resistant to apoptosis rescue irrespective of the stimulus. This was not linked to differences in proliferation because both CD27– and CD27+ B cells from CVID MB0 patients proliferated similarly to controls and CVID MB1 patients (Fig. 3c,d). IL-21 alone was able to rescue CD27– (16·9%) but not CD27+ B cells from spontaneous apoptosis (Figs 1a and 4). In spite of this, the addition of IL-21 down-modulated the protective effect of anti-CD40 (77·9 versus 75·9%, P < 0·01)

and CpG-ODN (71·4 versus 42·7%, P < 0·001) on CD27– B cells. In CD27+ B cells IL-21 tended to reduce the CpG-ODN rescue effect but increased the protective effect of anti-CD40 significantly (23·9 versus 42·8%, P < 0·05) (Figs 1a and selleck chemical 4b). IL-21 not only reverted the protective effect of anti-IgM on CD27– and CD27+ B cells, but in some cases even increased apoptosis above spontaneous baseline values (Fig. 1a and scatter-plots in Fig. 4). Similar results were obtained when we evaluated activation induced rescue from apoptosis on sorted CD27– and CD27+ B lymphocytes stimulated with the same stimuli (histograms in Fig. 1b,c). Moreover, we did not find increased CD27 expression when we stimulated CD27– B cells with any of the stimuli (dot-plots

in Fig. 1b), which validates the gating strategy when using purified total B cells. IL-21 modulates proliferation induced by co-stimulation on CD27– and CD27+ B cells. This effect has to be taken into account when analysing the apoptosis SAHA HDAC ic50 rate. Neither CD27– nor CD27+ B cells proliferated in response to anti-IgM combined with IL-21 (Table 2). However, both subpopulations proliferated in response to IL-21 with anti-CD40, although the proliferation index was higher in CD27+ B cells. Remarkably, IL-21 increased proliferation of CpG-ODN-activated Protirelin CD27– B cells but decreased proliferation of CpG-ODN-activated CD27+ B cells (Table 2).

In CD27+ B cells, IL-21 reduction of CpG-ODN apoptosis rescue is accompanied by a reduction in the proliferative response. In contrast, the increase in anti-CD40 apoptosis rescue is accompanied by a proliferation enhancement (Fig. 4b and Table 2). However, IL-21 reduction in apoptosis rescue induced by anti-CD40 or CpG-ODN on CD27– B cells is not due to a negative effect on proliferation (Fig. 4a and Table 2). Furthermore, in spite of the higher proliferative response induced by IL-21 combined with anti-CD40 or CpG-ODN on CD27+ versus CD27– B cells (Table 2), the rescue from apoptosis is not higher in CD27+ B cells for any of the stimulus (Fig. 4). Thus, although we cannot rule out that the effect of IL-21 on apoptosis is linked to proliferation, our results support the independence of these processes. IL-21 alone rescued both CVID MB0 and MB1 CD27– B cells similar to controls.

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