Many acute viral infections induce early T cell independent polyclonal B cell activation and production of antigen-specific IgA or IgG prior to the formation of classical T cell dependent B germinal centers. However, the necessary signals for initiation of early T cell independent class switch recombination (CSR) are not known. The objective of this application is to determine the essential pathways that drive T cell independent B cell activation and IgA CSR. We propose to utilize the murine model of rotavirus infection in which Peyer's patch (PP) T cell independent polyclonal B cell activation is induced within 48 hours after viral exposure followed by production of rotavirus- specific IgA. Rotavirus induced B cell activation in vitro requires dendritic cells. Rotavirus antigen is present in PP dendritic cells in vivo and these cells are activated at the time B cell activation occurs, suggesting that dendritic cells are important modulators of the B cell response to rotavirus. Our finding that B cell activation is dependent on the structural integrity of the rotavirus particles suggested the possibility that rotavirus induces dendritic cell responses as a result of pattern recognition. Preliminary experiments demonstrate that lack of MyD88 expression results in an ablation of B cell activation and substantial reduction in intestinal IgA levels. We find that a mutation in the B cell activating factor (BAFF) signaling pathway also results in the ablation of B cell activation, suggesting a potential link between MyD88 and BAFF. Several days following PP B cell activation there is an increase of TGF-2 expression and numbers of IgA+ B cells in the PP, suggesting TGF-2 also plays an important role in the PP polyclonal response to rotavirus. Therefore, we hypothesize that dendritic cells modulate the production of rapid T cell independent pathogen-specific antibody in the PP through induction of a polyclonal B cell response that results from MyD88, BAFF, and TGF-2 signaling independent of an antigen-specific BCR. We will test this hypothesis by using both in vitro and in vivo approaches to pursue the following three aims:
Aim 1. Determine whether rotavirus induces dendritic cell MyD88 signaling that drives B cell responses.
Aim 2. Determine whether MyD88 signaling results in BAFF and TGF-ss production.
Aim 3. Characterize the polyclonal B cell responses to rotavirus. The studies proposed in this application are of importance in addressing unanswered questions regarding the molecular mechanisms of T cell independent antibody induction in the intestine. This work will provide new insights into intestinal immune responses as well as contribute to improved mucosal vaccine strategies. Defining how pathogens induce rapid but specific antibody has tremendous potential to define the role of early antibody in limiting viral replication and dissemination and providing protection.
This proposal aims to understand how the intestinal antibody response to a viral pathogen is initiated and regulated by the host. Specifically, the earliest signals between dendritic cells and B cells required to induce B cells to produce viral specific antibodies will be investigated. Information gained will provide a more comprehensive understanding of intestinal immune regulation by defining novel signaling pathways and will be pertinent to development of therapeutics and mucosal vaccines.
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