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Abstract
Detection of antigen-specific memory B cells for immune monitoring requires their activation, and is commonly accomplished through stimulation with the TLR7/8 agonist R848 and IL-2. To this end, we evaluated whether addition of IL-21 would further enhance this TLR-driven stimulation approach; which it did not. More importantly, as most antigen-specific B cell responses are T cell-driven, we sought to devise a polyclonal B cell stimulation protocol that closely mimics T cell help. Herein, we report that the combination of agonistic anti-CD40, IL-4 and IL-21 affords polyclonal B cell stimulation that was comparable to R848 and IL-2 for detection of influenza-specific memory B cells. An additional advantage of anti-CD40, IL-4 and IL-21 stimulation is the selective activation of IgM+ memory B cells, as well as the elicitation of IgE+ ASC, which the former fails to do. Thereby, we introduce a protocol that mimics physiological B cell activation through helper T cells, including induction of all Ig classes, for immune monitoring of antigen-specific B cell memory.
Highlights
The adaptive immune system, on one hand, enables the host to develop long-term immunity to antigens; on the other hand, it allows for fine-tuning of effector functions that are required for successful defense against different challenges
While an increased abundance of IgM was observed with addition of IL-21 to R848 + IL-2 (Condition 3) by ELISA, it did not reach the level of significance (p = 0.059)
The data presented here suggest that IL-21 has no major effect on IgM, IgG and IgA production when combined with R848 and IL-2
Summary
The adaptive immune system, on one hand, enables the host to develop long-term immunity to antigens; on the other hand, it allows for fine-tuning of effector functions that are required for successful defense against different challenges. B cells switch to producing IgG and IgA antibodies that can penetrate most tissues of the body and are exported to mucosal surfaces, which extends immune surveillance to the entire body Under special circumstances, such as persistent antigen exposure, IgE antibodies are elicited and bind to mast cells and basophils, thereby sensitizing them for local histamine release at the site of antigen encounter. The B cell maintains an identical hypervariable region, which defines antibody specificity, but splices it with a downstream constant region defining a different class/subclass of immunoglobulin This process of gene rearrangement, called immunoglobulin class switching, is largely under the control of antigen-specific T helper (TH ) cells, and predominantly occurs within secondary lymphoid structures, such as the spleen and lymph nodes (reviewed in [2])
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