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While it is plausible that these potentially pathogenic B cells have a limited time frame in which they can undergo activation, our data indicate that mature high affinity DNA-reactive B cells that are expanded during B cell reconstitution remain part of the naïve repertoire for at least 1 month and can be activated to secrete autoantibody.
Taken together, these findings suggest that moLCs can efficiently be activated to secrete cytokines and express TLR4 via TLR signaling but not through E-cadherin crosslinking.
To be convinced that atypical MBCs could not be activated to secrete Abs we tried a number of conventional B cell stimulants.
A second stimulus, such as lipopolysaccaride (LPS), is generally required for GM-CSF or M-CSF-primed monocytes and macrophages to be activated to secrete significant levels of different cytokines.
JurkatαCEA-CIR-EGFP cells could be activated to secrete IL-2, upon antigen-mediated ligation of their binding domain by plastic immobilized CEA (iCEA) or membrane-bound CEA (mCEA) (Fig. 2).
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Upon contact with antigen, DCs are activated to secrete an array of cytokines and chemokines.
Previous investigations in mice suggested that NK cells are activated to secrete IFNγ by IL-12 released from APC in response to CD40L expression by iNKT cells [11], with a significant contribution also attributable to iNKT cell-derived IFNγ [41].
Notably, in response to either bacterial-derived GalA-GSL or αGalCer, the majority of NKT cells that were activated to secrete IL-4 and IFN-γ were highly labelled in vivo with CD45-PE.
Intriguingly, even in the absence of DCs and macrophages, around 20% of NKT cells were activated to secrete IFN-γ suggesting that other APCs such as B cells may also participate in NKT cell activation in the spleen.
Indeed, despite the decrease in circulating mDC, we found that serum IL-12 levels which is a Th1 type cytokine and mainly secreted by mDCs was significantly higher in the DM + UAP group compared with those in the DM, UAP groups and Normal controls, suggesting that mDCs in peripheral blood are activated to secrete cytokines in diabetic patients with UAP.
Moreover, at this time we detected a dramatic decrease in the percentage of NKT cells that were activated to secrete IFN-γ (23±5% versus 69±3%; Figure 7B, Supplementary Figure S5E), accompanied by a decrease in MFI for intracellular IFN-γ secreted by NKT cells (46±13% MFI reduction after CLL treatment versus untreated mice).
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Justyna Jupowicz-Kozak
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