Undergraduate Thesis Or Project
 

Method development in in vitro immune response

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  • The AhR is a ligand-activated transcription factor that mediates the potent immunosuppressive effects of TCDD. AhR-dependent changes in gene expression appear to alter the differentiation of CD4+ T cells to induce a population of CD4+CD25+ T regulatory cells (Tregs) that suppress the immune response. Ascertaining if and how the AhR induces Tregs is clinically relevant. Tregs act to suppress activation of the immune system, maintain immune system homeostasis, and tolerance to self. If the immunosuppressive functions of Tregs could be controlled via activation of the AhR for example, autoimmune diseases could potentially be treated, and the immune system could be prevented from rejecting transplanted organs. An in vitro model was designed to test the hypothesis that TCDD acts directly on the AhR in T cells to induce CD4+CD25+ Tregs. A method was developed to extract dendritic cells from the spleens of B6D2F1 mice. T cells were isolated from the spleens of C57Bl/6 AhR-/- and AhR+/+ mice. Splenic dendritic cells are potent activators of T cells. TCDD was added to cultures at concentrations of 1x10-8, 1x10-9, 1x10-10, and 1x10-11. TCDD had no notable effect on T cell viability or CD25 expression. TCDD did however increase the percentage of dividing CD4+ T cells that downregulated CD62L, an effect that was also seen in vivo on CD4+CD25+ T cells. These results suggest that TCDD acts directly on CD4+ T cells to alter CD62L expression, but further differentiation into Tregs was not observed. Future studies will examine other culture conditions that may support direct AhR-dependent induction of Tregs.
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