U01AI163062

Perforin 2 Controls Unconventional Cytokine Release from Mucosal APC

Project Summary

The understanding of how professional antigen presenting cell (APC) populations deliver cytokines to T cells, thereby shaping the pro-inflammatory vs. anti-inflammatory status of mucosal tissue, remains incomplete. Specifically, cytokines such as IL-33, which lack an N-terminal peptide sequence, are unable to access conventional protein secretion pathways. This has led to the prevailing view that cell death is responsible for IL-33 release.

This project builds upon exciting preliminary data demonstrating that mucosal conventional dendritic cell (CDC) subsets, in both humans and mice, express the transmembrane pore-forming protein perforin-2, which facilitates IL-33 secretion, at least in mice. In human CDC, perforin-2 expression is primarily found in an interferon regulatory factor 4 (IRF4) subset indicating the CDC2 lineage. In mouse CDC, perforin-2 is found in the CD103+ CDC1 subset known to express the transcription factors IRF8 and BATF3. Regardless of this lineage distinction, both human and mouse CD11C+ cells in the respiratory mucosa contain cytoplasmic IL-33 protein.

The data demonstrate that inhibition of perforin-2 activity prevents IL-33 release from CDC and inhibits the proliferative expansion of a poorly understood ST2+GATA3+FOXP3+TREG subset. Additionally, perforin-2 has been shown to regulate type 1 IFN signaling through controlling IFNAR responsiveness. Therefore, it is proposed that an important regulatory mechanism exists in humans and mice that is dependent upon mucosal APC expressing perforin-2.

This project aims to test the central hypothesis that APC require perforin-2 as an inducible plasma membrane conduit for unconventional cytokine delivery at the mucosal interface. Three specific aims will be investigated.

Specific Aim 1 (SA1) will determine whether perforin-2+ APC predict TREG subset abundance in sinonasal mucosa and define the transcriptional landscape of perforin-2+ APC.

Specific Aim 2 (SA2) will define the perforin-2 domains required for IL-33 release, the diversity of perforin-2-dependent secreted molecules, and the requisite intracellular trafficking machinery responsible for perforin-2 plasma membrane localization during APC-T cell interactions.

Specific Aim 3 (SA3) will directly test whether CDC1 and/or CDC2 subsets preferentially use perforin-2 for driving pathogen-specific T cell responses in mouse models of respiratory virus or helminth infection.

Taken together, this MIST project stands to uncover a new paradigm for understanding how CDC instruct immunity within the respiratory mucosa.

Trustees Of The University Of Pennsylvania

TO ASSIST PUBLIC AND PRIVATE NONPROFIT INSTITUTIONS AND INDIVIDUALS TO ESTABLISH, EXPAND AND IMPROVE BIOMEDICAL RESEARCH AND RESEARCH TRAINING IN INFECTIOUS DISEASES AND RELATED AREAS, TO CONDUCT DEVELOPMENTAL RESEARCH, TO PRODUCE AND TEST RESEARCH MATERIALS. TO ASSIST PUBLIC, PRIVATE AND COMMERCIAL INSTITUTIONS TO CONDUCT DEVELOPMENTAL RESEARCH, TO PRODUCE AND TEST RESEARCH MATERIALS, TO PROVIDE RESEARCH SERVICES AS REQUIRED BY THE AGENCY FOR PROGRAMS IN INFECTIOUS DISEASES, AND CONTROLLING DISEASE CAUSED BY INFECTIOUS OR PARASITIC AGENTS, ALLERGIC AND IMMUNOLOGIC DISEASES AND RELATED AREAS. PROJECTS RANGE FROM STUDIES OF MICROBIAL PHYSIOLOGY AND ANTIGENIC STRUCTURE TO COLLABORATIVE TRIALS OF EXPERIMENTAL DRUGS AND VACCINES, MECHANISMS OF RESISTANCE TO ANTIBIOTICS AS WELL AS RESEARCH DEALING WITH EPIDEMIOLOGICAL OBSERVATIONS IN HOSPITALIZED PATIENTS OR COMMUNITY POPULATIONS AND PROGRESS IN ALLERGIC AND IMMUNOLOGIC DISEASES. BECAUSE OF THIS DUAL FOCUS, THE PROGRAM ENCOMPASSES BOTH BASIC RESEARCH AND CLINICAL RESEARCH. SMALL BUSINESS INNOVATION RESEARCH (SBIR) PROGRAM EXPANDS AND IMPROVES PRIVATE SECTOR PARTICIPATION IN BIOMEDICAL RESEARCH. THE SBIR PROGRAM INTENDS TO INCREASE AND FACILITATE PRIVATE SECTOR COMMERCIALIZATION OF INNOVATIONS DERIVED FROM FEDERAL RESEARCH AND DEVELOPMENT, TO INCREASE SMALL BUSINESS PARTICIPATION IN FEDERAL RESEARCH AND DEVELOPMENT, AND TO FOSTER AND ENCOURAGE PARTICIPATION OF SOCIALLY AND ECONOMICALLY DISADVANTAGED SMALL BUSINESS CONCERNS AND WOMEN-OWNED SMALL BUSINESS CONCERNS IN TECHNOLOGICAL INNOVATION. THE SMALL BUSINESS TECHNOLOGY TRANSFER (STTR) PROGRAM STIMULATES AND FOSTERS SCIENTIFIC AND TECHNOLOGICAL INNOVATION THROUGH COOPERATIVE RESEARCH AND DEVELOPMENT CARRIED OUT BETWEEN SMALL BUSINESS CONCERNS AND RESEARCH INSTITUTIONS, TO FOSTER TECHNOLOGY TRANSFER BETWEEN SMALL BUSINESS CONCERNS AND RESEARCH INSTITUTIONS, TO INCREASE PRIVATE SECTOR COMMERCIALIZATION OF INNOVATIONS DERIVED FROM FEDERAL RESEARCH AND DEVELOPMENT, AND TO FOSTER AND ENCOURAGE PARTICIPATION OF SOCIALLY AND ECONOMICALLY DISADVANTAGED SMALL BUSINESS CONCERNS AND WOMEN-OWNED SMALL BUSINESS CONCERNS IN TECHNOLOGICAL INNOVATION. RESEARCH CAREER DEVELOPMENT AWARDS SUPPORT THE DEVELOPMENT OF SCIENTISTS DURING THE FORMATIVE STAGES OF THEIR CAREERS. INDIVIDUAL NATIONAL RESEARCH SERVICE AWARDS (NRSAS) ARE MADE DIRECTLY TO APPROVE APPLICANTS FOR RESEARCH TRAINING IN SPECIFIED BIOMEDICAL SHORTAGE AREAS. IN ADDITION, INSTITUTIONAL NATIONAL RESEARCH SERVICE AWARDS ARE MADE TO ENABLE INSTITUTIONS TO SELECT AND MAKE AWARDS TO INDIVIDUALS TO RECEIVE TRAINING UNDER THE AEGIS OF THEIR INSTITUTIONAL PROGRAM.

93.855 - Allergy and Infectious Diseases Research

National Institute of Allergy and Infectious Diseases (NIAID) [HHS - NIH]

Pennsylvania United States

State-Wide

Mucosal Immunology Studies Team (MIST) (U01 Clinical Trial Not Allowed). (RFA-AI-20-027)

Amendment Since initial award the End Date has been shortened from 05/31/26 to 11/30/24 and the total obligations have increased 424% from $455,000 to $2,384,327.