Multivesicular bodies in intestinal epithelial cells: responsible for MHC class II-restricted antigen processing and origin of exosomes.
In normal conditions intestinal epithelial cells (IECs) constitutively stimulate regulatory CD4(+) T cells. However, in Crohn's disease (CD), this major histocompatibility complex (MHC) class II-restricted antigen presentation results in stimulation of proinflammatory CD4(+) T cells. We hypothesized that these alternative functions might be mediated by differential sorting and processing of antigens into distinct MHC II-enriched compartments (MIICs). Accordingly, we analysed the endocytic pathways of lumenally applied ovalbumin (OVA) in IECs of the jejunum and ileum of wild-type (WT) and TNFDeltaARE/WT mice that develop a CD-resembling ileitis. Using quantitative reverse transcription polymerase chain reaction, we found that messenger RNA levels of interferon-gamma, tumour necrosis factor-alpha, interleukin-17 and interleukin-10 were significantly up-regulated in the inflamed ileum of TNFDeltaARE/WT mice, confirming CD-like inflammation. Fluorescence and immunoelectron microscopy revealed the presence of MHC II and invariant chain throughout the late endocytic compartments, with most molecules concentrated in the multivesicular bodies (MVB). OVA was targeted into MVB and, in contrast to other MIICs, accumulated in these structures within 120 min of exposure. The IEC-specific A33 antigen localized to internal vesicles of MVB and A33/class II-bearing exosomes were identified in intercellular spaces. Remarkably, the expression pattern of MHC II/invariant chain molecules and the trafficking of OVA were independent of mucosal inflammation and the specific region in the small intestine. MVB seem to be principally responsible for class II-associated antigen processing in IECs and to constitute the origin of MHC II-loaded exosomes. The distinctive functions of IECs in antigen presentation to CD4(+) T cells might arise as a result of differential processing within the MVB identified here.
Büning
Jürgen
J
Department of Internal Medicine I, University Hospital of Schleswig-Holstein, Lübeck, Germany. juergen.buening@uk-sh.de
von Smolinski
Dorthe
D
Tafazzoli
Kianush
K
Zimmer
Klaus-Peter
KP
Strobel
Stephan
S
Apostolaki
Maria
M
Kollias
George
G
Heath
Joan K
JK
Ludwig
Diether
D
Gebert
Andreas
A
D000818:Animals; D017951:Antigen Presentation; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis; D001692:Biological Transport; D003424:Crohn Disease / Q000276:immunology; D016207:Cytokines / Q000032:analysis / Q000235:genetics; D004195:Disease Models, Animal; D011992:Endosomes / Q000276:immunology* / Q000648:ultrastructure; D004847:Epithelial Cells / Q000276:immunology / Q000648:ultrastructure; D055354:Exosomes / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000032:analysis / Q000276:immunology*; D007082:Ileum; D007413:Intestinal Mucosa / Q000276:immunology* / Q000648:ultrastructure; D051379:Mice; D018345:Mice, Knockout; D016253:Microscopy, Immunoelectron; D010047:Ovalbumin; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D000944:Antigens, Differentiation, B-Lymphocyte; D016207:Cytokines; D000949:Histocompatibility Antigens Class II; D014409:Tumor Necrosis Factor-alpha; C045084:invariant chain; D010047:Ovalbumin