MHC class I antigen processing of an adenovirus CTL epitope is linked to the levels of immunoproteasomes in infected cells.
Proteasomes are the major source for the generation of peptides bound by MHC class I molecules. To study the functional relevance of the IFN-gamma-inducible proteasome subunits low molecular mass protein 2 (LMP2), LMP7, and mouse embryonal cell (MEC) ligand 1 in Ag processing and concomitantly that of immunoproteasomes, we established the tetracycline-regulated mouse cell line MEC217, allowing the titrable formation of immunoproteasomes. Infection of MEC217 cells with Adenovirus type 5 (Ad5) and analysis of Ag presentation with Ad5-specific CTL showed that cells containing immunoproteasomes processed the viral early 1B protein (E1B)-derived epitope E1B192-200 with increased efficiency, thus allowing a faster detection of viral entry in induced cells. Importantly, optimal CTL activation was already achieved at submaximal immunosubunit expression. In contrast, digestion of E1B-polypeptide with purified proteasomes in vitro yielded E1B192-200 at quantities that were proportional to the relative contents of immunosubunits. Our data provide evidence that the IFN-gamma-inducible proteasome subunits, when present at relatively low levels as at initial stages of infection, already increase the efficiency of antigenic peptide generation and thereby enhance MHC class I Ag processing in infected cells.
Sijts
A J
AJ
Institute of Biochemistry, Charité, Humboldt University, Berlin, Germany.
Standera
S
S
Toes
R E
RE
Ruppert
T
T
Beekman
N J
NJ
van Veelen
P A
PA
Ossendorp
F A
FA
Melief
C J
CJ
Kloetzel
P M
PM
D000260:Adenoviruses, Human / Q000235:genetics / Q000276:immunology*; D000276:Adjuvants, Immunologic / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D017951:Antigen Presentation / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000276:immunology* / Q000378:metabolism* / Q000502:physiology; D004306:Dose-Response Relationship, Immunologic; D004790:Enzyme Induction / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000378:metabolism*; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000096:biosynthesis / Q000276:immunology* / Q000378:metabolism* / Q000502:physiology; D010452:Peptide Biosynthesis / Q000276:immunology; D046988:Proteasome Endopeptidase Complex; D013602:T-Lymphocytes, Cytotoxic / Q000201:enzymology* / Q000276:immunology* / Q000378:metabolism / Q000821:virology; D013752:Tetracycline / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured
D000276:Adjuvants, Immunologic; D018984:Epitopes, T-Lymphocyte; D015395:Histocompatibility Antigens Class I; D009097:Multienzyme Complexes; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D013752:Tetracycline