Mechanisms for Initiation of Food Allergy Early in Life

Statement of Work: Bethany E. Perez White As outlined in Specific Aim 3, we are proposing to test the hypothesis that allergens act directly on keratinocytes, thereby inducing functional changes in these skin cells that contribute to food allergy. To test this, we will employ 3D cultures of primary human and mouse keratinocytes. In my laboratory and in the Skin Tissue Engineering and Morphology (STEM) Core in which I act as an Associate Director, we have extensive experience in the culture of primary epidermal keratinocytes derived from human and mouse. These models provide a unique system to study endogenous biological processes in a culture system that mimics the physiological architecture and function of epidermal tissue. Dr. Cook-Mills and I have worked together in the past, and have generated preliminary data integral to this R01 submission. My contributions shall include the development and preliminary culture of 3D skin equivalents as specified in Aim 3.C.1 and 3.C.2. I shall oversee sample preparation for histology/immunohistochemistry. Given my experience in keratinocyte differentiation, I shall advise on molecular mechanisms and signaling involved in this process. I shall also provide training to members of the Cook-Mills laboratory or participants on the proposed project in the culture of 3D skin equivalents