Date of Award

January 2017

Document Type

Thesis

Degree Name

Medical Doctor (MD)

Department

Medicine

First Advisor

Ruslan Medzhitov

Abstract

The aim of this project was to investigate the role of specialized epithelial sensors in allergic inflammation. We hypothesize that enterochromaffin cells play a pivotal role in the sensing of gut allergens, the induction of an inflammatory response to food allergens, and the execution of avoidance behaviors. Purported enterochromaffin cell line BON cells were grown and characterized by qPCR. BON cells were stimulated with ionomycin to induce degranulation, visualized by immunofluorescent microscopy, and analyzed by flow cytometry. For a model of type 2 inflammation, mice were infected with Nippostrongylus brasiliensis or sensitized to ovalbumin. Gut tissue from infected/sensitized and control mice was examined by immunofluorescent microscopy and intestinal epithelial cells were isolated and analyzed by flow cytometry. BON cells were found to be a heterogeneous population of cells that contain serotonin and chromogranin A. Candidate enterochromaffin cells in the intestine of mice could be identified by immunofluorescence and flow cytometry using indirect immunostaining for CD45+ 5-HT+ EpCAM+ and CD45+ CgA+ EpCAM+ cells. No gross change in enterochromaffin cell number was observed after infection with Nippostrongylus brasiliensis. Multiple protocols were compared to determine optimal study conditions. This work provides improved methods and insights into potential directions for further study.

Comments

This thesis is restricted to Yale network users only. This thesis is permanently embargoed from public release.

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