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PMID: 10662796 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Efficient generation of a hepatitis B virus cytotoxic T lymphocyte epitope requires the structural features of immunoproteasomes.

The Journal of experimental medicine ·Vol. 191 ·No. 3 ·2000-02-07 ·Pages 503-14

Sijts AJ, Ruppert T, Rehermann B, Schmidt M, Koszinowski U, Kloetzel PM

Abstract

Interferon (IFN)-gamma-induced cells express the proteasome subunits low molecular weight protein (LMP)2, LMP7, and MECL-1 (multicatalytic endopeptidase complex-like 1), leading to the formation of immunoproteasomes. Although these subunits are thought to optimize MHC class I antigen processing, the extent of their role and the mechanistic aspects involved remain unclear. Herein, we study the proteolytic generation of an human histocompatibility leukocyte antigen (HLA)-Aw68-restricted hepatitis B virus core antigen (HBcAg) cytotoxic T lymphocyte (CTL) epitope that is recognized by peripheral blood lymphocytes from patients with acute self-limited but not chronic hepatitis B virus (HBV). Immunological data suggest that IFN-gamma-induced rather than uninduced HeLa cells process and present the HBV CTL epitope upon infection with HBcAg-expressing vaccinia viruses. Analyses of 20S proteasome digests of synthetic polypeptides covering the antigenic HBcAg peptide demonstrate that only immunoproteasomes efficiently perform the cleavages needed for the liberation of this HBV CTL epitope. Although the concerted presence of the three immunosubunits appears essential, we find that both catalytically active LMP7 and inactive LMP7 T1A support CTL epitope generation. We conclude that LMP7 influences the structural features of 20S proteasomes, thereby enhancing the activity of the LMP2 and MECL-1 catalytic sites, which provide cleavage specificity. Thus, LMP7 incorporation is of greater functional importance for the generation of an HBV CTL epitope than cleavage specificity.

MeSH Terms
Amino Acid Sequence Animals Binding Sites Cell Line Cysteine Endopeptidases/chemistry Epitopes, T-Lymphocyte/chemistry,immunology HeLa Cells Hepatitis B/immunology Hepatitis B Core Antigens/chemistry,immunology Humans Interferon-gamma/pharmacology Kinetics Major Histocompatibility Complex Mice Mice, Inbred BALB C Molecular Sequence Data Multienzyme Complexes/chemistry Peptide Fragments/chemistry Proteasome Endopeptidase Complex Proteins Structure-Activity Relationship T-Lymphocytes, Cytotoxic/immunology Transfection Viral Matrix Proteins
Chemicals
EBV-associated membrane antigen, Epstein-Barr virus Epitopes, T-Lymphocyte Hepatitis B Core Antigens Multienzyme Complexes Peptide Fragments Proteins Viral Matrix Proteins Interferon-gamma Cysteine Endopeptidases LMP7 protein Proteasome Endopeptidase Complex
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Sijts A J
Institute of Biochemistry, Charité, Humboldt University Berlin, 10117 Berlin, Germany.
Ruppert T
Rehermann B
Schmidt M
Koszinowski U
Kloetzel P M
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Article Info
Journal
The Journal of experimental medicine
Abbr.
J Exp Med
ISSN
0022-1007
DOI
10.1084/jem.191.3.503
Published
2000-02-07
Pages
503-14
Language
English
Region
United States
NLM ID
2985109R
PMCID
PMC2195811
Subset
IM
External Links
PubMed source DOI: 10.1084/jem.191.3.503 PMC Full Text View journal details
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