Immunomodulatory activity of recombinant α-gliadin conjugated to cholera toxin in DQ8 transgenic mice

- An approach that downregulate the Th1-like cell response to wheat gliadin is proposed.
- A conjugate cholera toxin-recombinant gliadin modulates the T cell response to gliadin.
- Dendritic cells pulsed with the conjugate could be therapeutic in Celiac Disease.

Coeliac disease (CD) is characterized by an intestinal lesion sustained by an abnormal mucosal T-cell response to wheat gliadin. An immunological approach that is able to suppress this immune response is a perspective worth pursuing. Several strategies of antigen administration have been aimed at the downregulation of pathogenic T-cells. In particular, we previously reported a significant suppression of the systemic cell-mediated response toward wheat gliadin in DQ8 transgenic mice receiving nasally a recombinant α-gliadin. To gain further insight about the cellular mechanisms underlying the tolerogenic properties of this molecule, we analysed different preparations of the recombinant α-gliadin, alone or conjugated to the adjuvant cholera toxin (CT), by in vitro challenge with spleen CD4+ T cells from gliadin-sensitized DQ8 tg mice. We found that a partially purified preparation of recombinant α-gliadin (r-gliadin) induced a significantly higher production of IFN-γ than native gliadin as well as HPLC purified r-gliadin. Interestingly, r-gliadin, but not HPLC purified r-gliadin, stimulated the gliadin-specific expression of IL-10 in CD4+ T cells. No significant cytotoxic effect was induced by r-gliadin in MODE-K cells, a murine model of enterocytes. Notably, a conjugate CT-r-gliadin failed in stimulating IFN-γ, whereas IL-10 secretion was still induced in gliadin-specific CD4+ T cells. In conclusion, our results showed that DCs, pulsed with CT-r-gliadin in vitro, could modulate the ongoing Th1-like T cell response toward wheat gliadin. This finding provides new insight into the design of immunomodulatory protocols potentially useful for CD.