Supplementary Material for: Induction of Regulatory T Cells as a Novel Mechanism Underlying the Therapeutic Action of Kakkonto, a Traditional Japanese Herbal Medicine, in a Murine Food Allergy Model Yamamoto T. Fujiwara K. Tsubota Y. Kageyama-Yahara N. Hayashi S. Kadowaki M. 10.6084/m9.figshare.5129770.v1 https://karger.figshare.com/articles/dataset/Supplementary_Material_for_Induction_of_Regulatory_T_Cells_as_a_Novel_Mechanism_Underlying_the_Therapeutic_Action_of_Kakkonto_a_Traditional_Japanese_Herbal_Medicine_in_a_Murine_Food_Allergy_Model/5129770 <b><i>Background:</i></b> The number of patients with food allergy (FA) has dramatically increased. Although satisfactory drug therapies for FA are not available, we have found that kakkonto, a traditional Japanese herbal medicine, suppressed the occurrence of allergic symptoms in an FA mouse model. Thus, we investigated whether kakkonto could regulate the activation and differentiation of T cells in the colon. <b><i>Methods:</i></b> BALB/c mice were systemically sensitized and then orally challenged with ovalbumin. FA mice were orally treated with kakkonto. Lamina propria (LP) cells from their colons were isolated and analyzed. <b><i>Results:</i></b> Kakkonto significantly reduced the proportion of CD69<sup>+</sup> cells and the elevated helper T cell type 2-specific transcription factor GATA-3 mRNA expression in the LP CD4<sup>+</sup> T cells, showing that kakkonto has a suppressive effect on the activation and Th2 differentiation of LP effector CD4<sup>+</sup> T cells of the FA mouse colon. Furthermore, kakkonto significantly increased the proportion of Foxp3<sup>+</sup>CD4<sup>+</sup> regulatory T cells in the LP CD4<sup>+</sup> T cells of the FA mouse colon. Similarly, the number of Foxp3-positive cells was dramatically increased in the colonic mucosa of kakkonto-administered FA mice. However, the pharmacological effect and Foxp3<sup>+</sup>CD4<sup>+</sup> regulatory T cell-inducing ability of kakkonto were not attenuated by the administration of an anti-CD25 monoclonal antibody in the FA model. <b><i>Conclusions:</i></b> The induction of Foxp3<sup>+</sup>CD4<sup>+</sup>CD25<sup>-</sup> regulatory T cells in the colon as a novel mechanism underlying the therapeutic action of kakkonto could be utilized for the development of a novel anti-FA drug. 2016-04-21 00:00:00 Food allergy Regulatory T cells Traditional herbal medicine Kakkonto Intestinal mucosal immunity Animal models