Antigen processing by nardilysin and thimet oligopeptidase generates cytotoxic T cell epitopes


Cytotoxic T lymphocytes (CTLs) recognize peptides presented by HLA class I molecules on the cell surface. The C terminus of these CTL epitopes is considered to be produced by the proteasome. Here we demonstrate that the cytosolic endopeptidases nardilysin and thimet oligopeptidase (TOP) complemented proteasome activity. Nardilysin and TOP were required, either together or alone, for the generation of a tumor-specific CTL epitope from PRAME, an immunodominant CTL epitope from Epstein-Barr virus protein EBNA3C, and a clinically important epitope from the melanoma protein MART-1. TOP functioned as C-terminal trimming peptidase in antigen processing, and nardilysin contributed to both the C-terminal and N-terminal generation of CTL epitopes. By broadening the antigenic peptide repertoire, nardilysin and TOP strengthen the immune defense against intracellular pathogens and cancer.

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Figure 1: PRA(190–198) is an HLA-A3-presented CTL epitope with a proteasome-independent C terminus.
Figure 2: Nardilysin produces precursors of PRA(190–198) with C-terminal extension.
Figure 3: TOP produces the C terminus of the PRA(190–198) epitope.
Figure 4: Role of nardilysin in HLA class I antigen processing.
Figure 5: The epitope-generating trimming capacity of TOP.
Figure 6: TOP-dependent presentation by HLA-A2 of the MART-1 CTL epitope.

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We thank T. Dannenberg and K. Textoris-Taube for technical assistance. Supported by the Dutch Cancer Society (UL 2005-3245) and Stichting Vanderes.

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J.H.K. conceived of the study, coordinated the work, designed, did and analyzed most experiments and wrote the manuscript with major input from C.J.M.M. and minor input from other authors; C.J.M.M., F.O. and P.A.v.V. provided intellectual input; S.K. did the experiments and analyses in Figure 2b and immunoblot analysis; K.M.C., L.B.H. and A. Prat contributed to the experiments about nardilysin; D.W.R., K.R., U.S. and A. Paschen contributed to the experiments about TOP; P.M.K., U.S. and B.T. contributed to the experiments about TPPII; H.S.O. and P.F.v.S. contributed to the experiments about the proteasome; J.N. and T.v.H. contributed to the experiments about TAP; N.v.M., U.S., A. Paschen, S.L.G. and J.M.B. contributed to CTL experiments; J.W.D., F.O., J.N., S.K. and W.E.B. contributed to substrate design and synthesis; S.A.B.-V. and K.L.M.C.F. contributed to the molecular biology; A.M. and I.I.N.D. made HLA class I mAbs and cell lines expressing a single HLA class I allele; and P.A.v.V. and A.d.R. did mass spectrometry.

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Correspondence to Jan H Kessler.

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Kessler, J., Khan, S., Seifert, U. et al. Antigen processing by nardilysin and thimet oligopeptidase generates cytotoxic T cell epitopes. Nat Immunol 12, 45–53 (2011).

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