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

Alice J A M Sijts, Thomas Ruppert, Barbara Rehermann, Marion Schmidt, Ulrich Koszinowski, Peter M. Kloetzel

Research output: Contribution to journalArticle

125 Citations (Scopus)

Abstract

Interferon (IFN)-γ-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-γ-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.

Original languageEnglish (US)
Pages (from-to)503-513
Number of pages11
JournalJournal of Experimental Medicine
Volume191
Issue number3
DOIs
StatePublished - Feb 7 2000
Externally publishedYes

Fingerprint

T-Lymphocyte Epitopes
Cytotoxic T-Lymphocytes
Hepatitis B virus
Proteasome Endopeptidase Complex
Hepatitis B Core Antigens
Interferons
Histocompatibility Antigens Class I
Peptides
Histocompatibility Antigens
Vaccinia virus
Chronic Hepatitis B
Antigen Presentation
HLA Antigens
HeLa Cells
Catalytic Domain
Molecular Weight
Lymphocytes
Infection

Keywords

  • Antigen processing
  • Hepatitis B virus
  • Immunoproteasome
  • LMP7T1A
  • MHC class I

ASJC Scopus subject areas

  • Immunology

Cite this

Efficient generation of a hepatitis B virus cytotoxic T lymphocyte epitope requires the structural features of immunoproteasomes. / Sijts, Alice J A M; Ruppert, Thomas; Rehermann, Barbara; Schmidt, Marion; Koszinowski, Ulrich; Kloetzel, Peter M.

In: Journal of Experimental Medicine, Vol. 191, No. 3, 07.02.2000, p. 503-513.

Research output: Contribution to journalArticle

Sijts, Alice J A M ; Ruppert, Thomas ; Rehermann, Barbara ; Schmidt, Marion ; Koszinowski, Ulrich ; Kloetzel, Peter M. / Efficient generation of a hepatitis B virus cytotoxic T lymphocyte epitope requires the structural features of immunoproteasomes. In: Journal of Experimental Medicine. 2000 ; Vol. 191, No. 3. pp. 503-513.
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