Commensal microbes have profound effects on human health. However, the nature of the commensal microbes that stimulate and regulate the immune system of the host is largely unknown. Therefore, the mechanisms by which such microbes engage host immunity are unknown. However, genetic tools to study immune effects of the microbiota are currently not available. Moreover, it is unclear how many of the microbiota species are amenable to genetic manipulation. Generation of TCR transgenic mice against individual commensal microbes is an important approach to study such immune responses. However, generation of individual TCR Tg mice against the hundreds of microbiota species is a monumental task. Here, we propose an alternative approach, in which we will use a single TCR Tg mice to track T cell responses to most of the microbiota and identify immunomodulatory commensal species. To this goal we developed an approach for Metagenomic Alteration of Gut microbiome by In situ Conjugation, or MAGIC. MAGIC leads to expression of exogenous proteins in a broad range of microbiota species simultaneously. We will use this system to 1) create a biobank of genetically-modified commensal microbes that express immunogenic proteins; 2) identify and isolate immunogenic commensals and 3) characterize the generated T cell responses.

Public Health Relevance

We propose to identify novel immunomodulatory commensals that activate host T cells at steady state. We will apply a high-throughput system that we developed to express an exogenous T cell antigen throughout the gut microbiota in an unbiased manner. We will then isolate engineered microbe and test their ability to activate host T cells in vivo to identify immunogenic species. We will also generate a biobank of genetically modified commensal species from both mice and humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AI146817-02
Application #
9941032
Study Section
Gastrointestinal Mucosal Pathobiology Study Section (GMPB)
Program Officer
Rothermel, Annette L
Project Start
2019-06-03
Project End
2021-05-31
Budget Start
2020-06-01
Budget End
2021-05-31
Support Year
2
Fiscal Year
2020
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
621889815
City
New York
State
NY
Country
United States
Zip Code
10032