Abstract
Several strategies that increase proteasomal degradation of antigen have been shown to improve MHC class I presentation of antigen. Because recent studies have demonstrated that the centrosome is a subcellular compartment rich in proteasomes, we hypothesized that targeting a tumor antigen to centrosomal compartments would enhance both the MHC class I presentation of antigen and the vaccine potency. We, therefore, created a chimera of gamma-tubulin, an established centrosomal marker, with a model tumor antigen, human papillomavirus type 16 (HPV-16) E7, in a DNA vaccine. The linkage of gamma-tubulin to E7-targeted antigen to centrosomal compartments, resulted in enhanced MHC class I presentation of E7, and led to a marked increase in the number of E7-specific CD8(+) T-cell precursors as well as a potent CD4-independent antitumor effect against an E7-expressing tumor cell line, TC-1. In addition, vaccination with gamma-tubulin/E7 DNA in transporter associated with antigen presentation (TAP)-1-knockout mice revealed that the enhancement of E7-specific CD8(+) T-cell immune responses is TAP-1-dependent. Our data suggest that the centrosome may be an important locus for MHC class I antigen processing and that targeting antigen to the centrosome can improve DNA vaccine potency.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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ATP Binding Cassette Transporter, Subfamily B, Member 2
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ATP-Binding Cassette Transporters / genetics
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ATP-Binding Cassette Transporters / immunology
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Animals
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Antigen Presentation / immunology*
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CD4-Positive T-Lymphocytes / immunology
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CD8-Positive T-Lymphocytes / immunology
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Cancer Vaccines / genetics
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Cancer Vaccines / immunology*
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Centrosome / immunology*
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Centrosome / metabolism
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Chimerin Proteins / genetics
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Chimerin Proteins / immunology
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Female
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Histocompatibility Antigens Class I / immunology*
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Lung Neoplasms / immunology*
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Lung Neoplasms / prevention & control
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Oncogene Proteins, Viral / genetics
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Oncogene Proteins, Viral / immunology*
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Oncogene Proteins, Viral / metabolism
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Papillomavirus E7 Proteins
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Transfection
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Tubulin / genetics
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Tubulin / immunology*
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Tubulin / metabolism
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Vaccines, DNA / genetics
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Vaccines, DNA / immunology*
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Vaccines, DNA / metabolism
Substances
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ATP Binding Cassette Transporter, Subfamily B, Member 2
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ATP-Binding Cassette Transporters
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Cancer Vaccines
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Chimerin Proteins
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Histocompatibility Antigens Class I
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Oncogene Proteins, Viral
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Papillomavirus E7 Proteins
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TAP1 protein, human
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Tap1 protein, mouse
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Tubulin
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Vaccines, DNA
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oncogene protein E7, Human papillomavirus type 16