Preparation of Protein-containing Extracts from Microbiota-rich Intestinal Contents
Patricia Dillenburg-Pilla,
Carlos Zárate-Bladés,
Phyllis Silver,
Reiko Horai,
Rachel Caspi
Affiliations
Patricia Dillenburg-Pilla
Department of Cell Biology, Embryology and Genetics, Federal University of Santa Catarina, Florianopolis, Santa Catarina, BrazilOral and Pharyngeal Cancer Branch, National Institutes of Dental and Craniofacial Research, National Institutes of Health, NIH, Bethesda, MD, USA
Carlos Zárate-Bladés
Laboratory of Immunoregulation, Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florianopolis, Santa Catarina, BrazilLaboratory of Immunology, National Eye Institute, NIH, Bethesda, MD, USA
Phyllis Silver
Laboratory of Immunology, National Eye Institute, NIH, Bethesda, MD, USA
Reiko Horai
Laboratory of Immunology, National Eye Institute, NIH, Bethesda, MD, USA
Rachel Caspi
Laboratory of Immunology, National Eye Institute, NIH, Bethesda, MD, USA
The contribution of microbiota in regulating multiple physiological and pathological host responses has been studied intensively in recent years. Evidence suggests that commensal microbiota can directly modulate different populations of cells of the immune system (e.g., Ivanov et al., 2008; Atarashi et al., 2011). Recently, we showed that protein extracts from gut commensal microbiota can activate retina-specific T cells, allowing these autoreactive T cells to then break through the blood-retinal barrier and trigger autoimmune uveitis in the recipient (Horai et al., 2015). The protocol below describes the method to prepare intestinal protein-rich extracts that can be used in various in vitro and in vivo immunological studies.
