Modulation of the Immune Response Using Dendritic Cell-Derived Exosomes
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Initial studies in our laboratory were focused on the use of dendritic cells (DC) genetically modified to express Th2-derived
cytokines (i.e., interleukin [IL]-4 and IL-10) or apoptotic proteins (i.e., Fas Ligand [FasL]) to reduce inflammation in a
mouse model of experimentally induced arthritis. Exosomes are nano-sized vesicles (40–100 nm diameter) released by different
cell types, including DC, that contain many of the proteins thought to be involved in regulating the immune response. We have
demonstrated that exosomes derived from immature DC treated with immunomodulatory cytokines (i.e., IL-10, IL-4) are able to
inhibit inflammation in a murine footpad model of delayed-type hypersensitivity (DTH) and reduce the severity of established
collagen-induced arthritis (CIA). In fact, the exosomes were as therapeutic as the parental DC. Because purified DC-derived
exosomes are very stable vesicles, they may be a better approach for future treatment of arthritis and other autoimmune disorders
than the more unstable DC. In this chapter we detail a protocol for preparing the exosomes produced by murine bone marrow-derived
DC. We also review methods to assess the purity and concentration of purified exosomes, by using electron microscopy, Western
blot analysis, and flow cytometry. Finally, we describe methods to assess the function of exosomes in vitro, using the mixed
leukocytes reaction, and in vivo by means of DTH and an experimental model of CIA.
Affiliation(s): (2) Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA
(3) Department of Surgery & Thomas E. Stazl Transplantation Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA
(3) Department of Surgery & Thomas E. Stazl Transplantation Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA
Book Title: Immunological Tolerance: Methods and Protocols
Series: Methods in Molecular Biology | Volume: 380 | Pub. Date: Jun-08-2007 | Page Range: 443-455 | DOI: 10.1007/978-1-59745-395-0_28
Subject: Immunology
Key Words: Rheumatoid arthritis - exosomes - autoimmune disease - dendritic cells - collagen-induced arthritis - delayed-type hypersensitivity
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