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Identification of a gap junction communication pathway critical in innate immunity

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dc.contributor.advisor Martin L. Yarmush. en_US
dc.contributor.author Patel, Suraj Jagdish en_US
dc.contributor.other Harvard University--MIT Division of Health Sciences and Technology. en_US
dc.date.accessioned 2011-04-25T16:12:45Z
dc.date.available 2011-04-25T16:12:45Z
dc.date.copyright 2010 en_US
dc.date.issued 2010 en_US
dc.identifier.uri http://hdl.handle.net/1721.1/62520
dc.description Thesis (Ph. D.)--Harvard-MIT Division of Health Sciences and Technology, 2010. en_US
dc.description Page 84 blank. Cataloged from PDF version of thesis. en_US
dc.description Includes bibliographical references. en_US
dc.description.abstract The innate immune system is the first line of host defense, and its ability to propagate antimicrobial and inflammatory signals from the cellular microenvironment to the tissue at-large is critical for survival. In a remarkably complex microenvironment, cells are constantly processing external cues, initiating convoluted intracellular signaling cascades, and interacting with neighboring cells to generate a global, unified response. At the onset of infection or sterile injury, individual cells sense danger or damage signals and elicit innate immune responses that spread from the challenged cells to surrounding cells, thereby establishing an overall inflammatory state. However, little is known about how these dynamic spatiotemporal responses unfold. Through the use GFP reporters, in vitro transplant coculture systems, and in vivo models of infection and sterile injury, this thesis describes identification of a gap junction intercellular communication pathway for amplifying immune and inflammatory responses, and demonstrates its importance in host innate immunity. The first section describes development of stable GFP reporters to study the spatiotemporal activation patterns of two key transcription factors in inflammation and innate immunity: Nuclear factor-KappaB (NFKB) and Interferon regulatory factor 3 (IRF3). Stimulation of NFKB-GFP reporters resulted in a spatially homogeneous pattern of activation, found to be largely mediated by paracrine action of the pro-inflammatory cytokine TNFa. In contrast, the activation of IRF3 was spatially heterogeneous, resulting in the formation of multicellular colonies of activated cells in an otherwise latent background. This pattern of activation was demonstrated to be dependent on cell-cell contact mediated communication between neighboring cells, and not on paracrine signaling. The second section describes the discovery of a gap junction intercellular communication pathway responsible for the formation of IRF3 active colonies in response to immune activation. Cell sorting and gene expression profiling revealed that the activated reporter colonies, collectively, serve as the major source of critical antimicrobial and inflammatory cytokines. Using in vitro transplant coculture systems, colony formation was found to be dependent on gap junction communication. Blocking gap junctions with genetic specificity severely compromised the innate immune system's ability to mount antiviral and inflammatory responses. The third section illustrates an application of the gap junction-induced amplification of innate immunity phenomenon in an animal model of sterile injury. Drug-induced liver injury was shown to be dependent on gap junction communication for amplifying sterile inflammatory signals. Mice deficient in hepatic gap junction protein connexin 32 (Cx32) were protected against liver damage, inflammation, and death in response to hepatotoxic drugs. Co-administration of a selective pharmacologic Cx32 inhibitor with hepatotoxic drugs significantly limited hepatocyte damage and sterile inflammation, and completely abrogated mortality. These finds suggests that co-formulation of gap junction inhibitors with hepatotoxic drugs may prevent liver failure in humans, and potentially limit other forms of sterile injury. In summary, this thesis demonstrates the development of novel tools for investigating the spatiotemporal dynamics of cellular responses, describes how these tools were utilized to discover a basic gap junction communication pathway critical in innate immunity, and provides evidence for the clinical relevance of this pathway in sterile inflammatory injury. en_US
dc.description.statementofresponsibility by Suraj Jagdish Patel. en_US
dc.format.extent 84 p. en_US
dc.language.iso eng en_US
dc.publisher Massachusetts Institute of Technology en_US
dc.rights M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission. en_US
dc.rights.uri http://dspace.mit.edu/handle/1721.1/7582 en_US
dc.subject Harvard University--MIT Division of Health Sciences and Technology. en_US
dc.title Identification of a gap junction communication pathway critical in innate immunity en_US
dc.type Thesis en_US
dc.description.degree Ph.D. en_US
dc.contributor.department Harvard University--MIT Division of Health Sciences and Technology. en_US
dc.identifier.oclc 712654706 en_US


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