33 results on '"Yewdell, Jonathan W."'
Search Results
2. Terminal Deoxynucleotidyl Transferase Establishes andBroadens Antiviral CD8+ T Cell Immunodominance Hierarchies1
- Author
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Haeryfar, S M Mansour, Hickman, Heather D., Irvine, Kari R, Tscharke, David, Bennink, Jack R, Yewdell, Jonathan W, Haeryfar, S M Mansour, Hickman, Heather D., Irvine, Kari R, Tscharke, David, Bennink, Jack R, and Yewdell, Jonathan W
- Abstract
The action of TdT on mouse TCR genes accounts for ∼90% of T cell repertoire diversity. We report that in TdT -/- mice, total T CD8+ responses to influenza and vaccinia viruses are reduced by ∼30% relative to wild-type mice. We find that T CD8+ respons
- Published
- 2008
3. Regulatory T cells suppress CD8 + T cell responses induced by direct priming and cross-priming and moderate immunodominance disparities
- Author
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Haeryfar, S M Mansour, DiPaolo, R J, Tscharke, David, Bennink, Jack R, Yewdell, Jonathan W, Haeryfar, S M Mansour, DiPaolo, R J, Tscharke, David, Bennink, Jack R, and Yewdell, Jonathan W
- Abstract
Little is known regarding the participation of CD4+CD25 + regulatory T cells (Treg) in TCD8+ responses. In this study, we show that Treg depletion via treatment with anti-CD25 mAb (PC61) significantly enhances TCD8+ responses to influenza A virus, vaccinia virus, and SV40-transformed cells induced by either direct priming or cross-priming. PC61 did not enhance T CD8+ responses in CD4-deficient mice, providing the initial demonstration that PC61 acts on a subset of TCD4+, and not on other cells that express either CD25 or a fortuitously cross-reactive Ag. We further show that Treg selectively suppress responses to the most immunodominant TCD8+ determinants in the three systems examined. Therefore, Treg influence T CD8 immunodominance hierarchies by moderating disparities in responses to different determinants.
- Published
- 2005
4. Mixed Proteasomes Function To Increase Viral Peptide Diversity and Broaden Antiviral CD8+ T Cell Responses.
- Author
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Zanker, Damien, Waithman, Jason, Yewdell, Jonathan W., and Weisan Chen
- Subjects
- *
PROTEASOMES , *PEPTIDES , *VIRAL proteins , *T cells , *PROTEOLYTIC enzymes , *CYTOKINES , *INFLUENZA A virus , *PHYSIOLOGY - Abstract
The three proteasome subunits with proteolytic activity are encoded by standard or immunoproteasome genes. Many proteasomes expressed by normal cells and cells exposed to cytokines are "mixed", that is, contain both standard and immunoproteasome subunits. Using a panel of 38 defined influenza A virus-derived epitopes recognized by C57BL/6 mouse CD8+ T cells, we used mice with targeted disruption of ß1i, ß2i, or ß5i/ß2i genes to examine the contribution of mixed proteasomes to the immunodominance hierarchy of antiviral CD8+ T cells. We show that each immunoproteasome subunit has large effects on the primary and recall immunodominance hierarchies due to modulating both the available T cell repertoire and generation of individual epitopes as determined both biochemically and kinetically in Ag presentation assays. These findings indicate that mixed proteasomes function to enhance the diversity of peptides and support a broad CD8+ T cell response. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
5. Cutting Edge: Myosin 18A Is a Novel Checkpoint Regulator in B Cell Differentiation and Antibody-Mediated Immunity.
- Author
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Cheung, Michael B., Enyindah-Asonye, Gospel, Ken Matsui, Kosik, Ivan, Dvorina, Nina, Baldwin III, William M., Yewdell, Jonathan W., and Gupta, Neetu
- Subjects
- *
B cell differentiation , *BONE marrow cells , *MYOSIN , *B cells , *BONE marrow - Abstract
We investigated the function of the newly discovered myosin family protein myosin 18A (Myo18A) in Ab-mediated immunity by generating B cell-conditional Myo18Adeficient mice. Myo18A deficiency led to expansion of bone marrow progenitor B cells and mature B cells in secondary lymphoid organs. Myo18A-deficient mice displayed serum IgM hyperglobulinemia and increased splenic IgM-secreting cells, with older mice switching to IgG1 hyperglobulinemia and autoantibody development. Immunization of Myo18A-deficient mice with inactivated influenza virus led to development of more potent neutralizing Abs against the major Ag hemagglutinin, associated with persistent accumulation of Ag-specific germinal center B cells and more Ag-specific bone marrow plasma cells. In vitro stimulation with TLR7 and BCR ligands revealed a greater ability of Myo18A-deficient B cells to differentiate into Ab-secreting cells, associated with higher AID and Blimp-1 expression. Overall, our study demonstrates that Myo18A is a novel negative regulator of B cell homeostasis, differentiation, and humoral immunity. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
6. Influenza A Virus Infection Induces Viral and Cellular Defective Ribosomal Products Encoded by Alternative Reading Framesd.
- Author
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Zanker, Damien J., Oveissi, Sara, Tscharke, David C., Duan, Mubing, Siyuan Wan, Xiaomu Zhang, Kun Xiao, Mifsud, Nicole A., Gibbs, James, Izzard, Lenny, Dlugolenski, Daniel, Faou, Pierre, Laurie, Karen L., Vigneron, Nathalie, Barr, Ian G., Stambas, John, Van den Eynde, Benoî J., Bennink, Jack R., Yewdell, Jonathan W., and Weisan Chen
- Subjects
- *
VIRUS diseases , *INFLUENZA A virus , *MANUFACTURING defects , *CELLULAR recognition , *T cells - Abstract
The importance of antiviral CD8+ T cell recognition of alternative reading frame (ARF)-derived peptides is uncertain. In this study, we describe an epitope (NS1-ARF21-8) present in a predicted 14-residue peptide encoded by the +1 register of NS1 mRNA in the influenza A virus (IAV). NS1-ARF21-8 elicits a robust, highly functional CD8+ T cell response in IAV-infected BALB/c mice. NS1- ARF21-8 is presented from unspliced NS mRNA, likely from downstream initiation on a Met residue that comprises the P1 position of NS1-ARF21-8. Derived from a 14-residue peptide with no apparent biological function and negligible impacts on IAV infection, infectivity, and pathogenicity, NS1-ARF21-8 provides a clear demonstration of how immunosurveillance exploits natural errors in protein translation to provide antiviral immunity. We further show that IAV infection enhances a model cellular ARF translation, which potentially has important implications for virus-induced autoimmunity. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
7. Influenza A Virus Negative Strand RNA Is Translated for CD8+ T Cell Immunosurveillance.
- Author
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Hickman, Heather D., Mays, Jacqueline W., Gibbs, James, Kosik, Ivan, Magádan, Javier G., Das, Suman, Reynoso, Glennys V., Ngudiankama, Barbara F., JiaJie Wei, Shannon, John P., McManus, Daniel, Yewdell, Jonathan W., and Kazuyo Takeda
- Subjects
- *
T cells , *INFLUENZA A virus , *BIOSYNTHESIS , *NEURAMINIDASE , *RNA viruses - Abstract
Probing the limits of CD8+ T cell immunosurveillance, we inserted the SIINFEKL peptide into influenza A virus (IAV)--negative strand gene segments. Although IAV genomic RNA is considered noncoding, there is a conserved, relatively long open reading frame present in segment 8, encoding a potential protein termed NEG8. The biosynthesis of NEG8 from IAV has yet to be demonstrated. Although we failed to detect NEG8 protein expression in IAV-infected mouse cells, cell surface Kb--SIINFEKL complexes are generated when SIINFEKL is genetically appended to the predicted C terminus of NEG8, as shown by activation of OT-I T cells in vitro and in vivo. Moreover, recombinant IAV encoding of SIINFEKL embedded in the negative strand of the neuraminidase-stalk coding sequence also activates OT-I T cells in mice. Together, our findings demonstrate both the translation of sequences on the negative strand of a single-stranded RNAvirus and its relevance in antiviral immunosurveillance. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
8. Varied Role of Ubiquitylation in Generating MHC Class I Peptide Ligands.
- Author
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Jiajie Wei, Damien Zanker, Di Carluccio, Anthony R., Smelkinson, Margery G., Kazuyo Taked, Seedhom, Mina O., Dersh, Devin, Gibbs, James S., Ning Yang, Jadhav, Ajit, Weisan Chen, and Yewdell, Jonathan W.
- Subjects
- *
UBIQUITINATION , *PEPTIDES , *T cells , *CELL lines , *CD8 antigen - Abstract
CD8+ T cell immunosurveillance is based on recognizing oligopeptides presented by MHC class I molecules. Despite decades of study, the importance of protein ubiquitylation to peptide generation remains uncertain. In this study, we examined the ability of MLN7243, a recently described ubiquitin-activating enzyme E1 inhibitor, to block overall cytosolic peptide generation and generation of specific peptides from vaccinia- and influenza A virus-encoded proteins. We show that MLN7243 rapidly inhibits ubiquitylation in a variety of cell lines and can profoundly reduce the generation of cytosolic peptides. Kinetic analysis of specific peptide generation reveals that ubiquitylation of defective ribosomal products is rate limiting in generating class I peptide complexes. More generally, our findings demonstrate that the requirement for ubiquitylation in MHC class I-restricted Ag processing varies with class I allomorph, cell type, source protein, and peptide context. Thus, ubiquitin-dependent and -independent pathways robustly contribute to MHC class I-based immunosurveillance. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
9. Protein Translation Activity: A New Measure of Host Immune Cell Activation.
- Author
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Seedhom, Mina O., Hickman, Heather D., Jiajie Wei, David, Alexandre, and Yewdell, Jonathan W.
- Subjects
- *
PROTEIN transport , *CELL populations , *T cells , *VIRUS diseases , *ACTIVATION (Chemistry) , *IMMUNE response , *PATHOGENIC microorganisms - Abstract
We describe the in vivo ribopuromycylation (RPM) method, which uses a puromycin-specific Ab to fluorescently label ribosomebound puromycylated nascent chains, enabling measurement of translational activity via immunohistochemistry or flow cytometry. Tissue staining provides a unique view of virus-induced activation of adaptive, innate, and stromal immune cells.RPMflow precisely quantitates virus-induced activation of lymphocytes and innate immune cells, and it provides a unique measure of immune cell deactivation and quiescence. Using RPM we find that high endothelial cells in draining lymph nodes rapidly increase translation in the first day of vaccinia virus infection. We also find a population of constitutively activated splenic T cells in naive mice and further that most bone marrow T cells activate 3 d after vaccinia virus infection. Bone marrow T cell activation is nonspecific, IL-12- dependent, and induces innate memory T cell phenotypic markers. Thus, RPM measures translational activity to uniquely identify cell populations that participate in the immune response to pathogens, other foreign substances, and autoantigens. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
10. Defining Viral Defective Ribosomal Products: Standard and Alternative Translation Initiation Events Generate a Common Peptide from Influenza A Virus M2 and Ml mRNAs.
- Author
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Ning Yang, Gibbs, James S., Hickman, Heather D., Reynoso, Glennys V., Ghosh, Arun K., Bennink, Jack R., and Yewdell, Jonathan W.
- Subjects
- *
RIBOSOMAL proteins , *GENETIC translation , *INFLUENZA A virus , *MESSENGER RNA , *GENETIC engineering - Abstract
Influenza A virus gene segment 7 encodes two proteins: the M1 protein translated from unspliced mRNA and the M2 protein produced by mRNA splicing and largely encoded by the M1 +1 reading frame. To better understand the generation of defective ribosomal products relevant to MHC class I Ag presentation, we engineered influenza A virus gene segment 7 to encode the model H-2 Kb class I peptide ligand SIINFEKL at the M2 protein C terminus. Remarkably, after treating virus-infected cells with the RNA splicing inhibitor spliceostatin A to prevent M2 mRNA generation, Kb-SIINFEKL complexes were still presented on the cell surface at levels ≤60% of untreated cells. Three key findings indicate that SIINFEKL is produced by cytoplasmic translation of unspliced M1 mRNA initiating at CUG codons within the +1 reading frame: 1) synonymous mutation of CUG codons in the M2-reading frame reduced Kb-SIINFEKL generation; 2) Kb-SIINFEKL generation was not affected by drug-mediated inhibition of AUG-initiated M1 synthesis; and 3) Kb-SIINFEKL was generated in vitro and in vivofrom mRNA synthesized in the cytoplasm by vaccinia virus, and hence cannot be spliced. These findings define a viral defective ribosomal product generated by cytoplasmic noncanonical translation and demonstrate the participation of CUG-codon-based translation initiation in pathogen immunosurveillance. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
11. Distinct Pathways Generate Peptides from Defective Ribosomal Products for CD8+ T Cell Immunosurveillance.
- Author
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Dolan, Brian P., Li, Lily, Veltri, Charles A., Ireland, Chris M., Bennink, Jack R., and Yewdell, Jonathan W.
- Subjects
- *
MAJOR histocompatibility complex , *PEPTIDES , *T cells , *IMMUNOLOGY , *ENZYMES - Abstract
To understand better the endogenous sources of MHC class I peptide ligands, we generated an antigenic reporter protein whose degradation is rapidly and reversibly controlled with Shield-1, a cell-permeant drug. Using this system, we demonstrate that defective ribosomal products (DRiPs) represent a major and highly efficient source of peptides and are completely resistant to our attempts to stabilize the protein. Although peptides also derive from nascent Shield-1-sensitive proteins and "retirees" created by Shield-1 withdrawal, these are much less efficient sources on a molar basis. We use this system to identify two drugs-each known to inhibit polyubiquitin chain disassembly-that selectively inhibit presentation of Shield-1-resistant DRiPs. These findings provide the initial evidence for distinct biochemical pathways for presentation of DRiPs versus retirees and implicate polyubiquitin chain disassembly or the actions of deubiquitylating enzymes as playing an important role in DRiP presentation. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
12. MLN4924 Inhibits Defective Ribosomal Product Antigen Presentation Independently of Direct NEDDylation of Protein Antigens.
- Author
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Vijayasimha K, Leestemaker-Palmer AL, Gibbs JS, Yewdell JW, and Dolan BP
- Subjects
- Animals, CD8-Positive T-Lymphocytes metabolism, Cyclopentanes, Mice, NEDD8 Protein metabolism, Proteins, Pyrimidines, Ubiquitin metabolism, Ubiquitins metabolism, Antigen Presentation, Proteasome Endopeptidase Complex metabolism
- Abstract
Successful direct MHC class I Ag presentation is dependent on the protein degradation machinery of the cell to generate antigenic peptides that can be loaded onto MHC class I molecules for surveillance by CD8
+ T cells of the immune system. Most often this process involves the ubiquitin (Ub)-proteasome system; however, other Ub-like proteins have also been implicated in protein degradation and direct Ag presentation. In this article, we examine the role of neuronal precursor cell-expressed developmentally downregulated protein 8 (NEDD8) in direct Ag presentation in mouse cells. NEDD8 is the Ub-like protein with highest similarity to Ub, and fusion of NEDD8 to the N terminus of a target protein can lead to the degradation of target proteins. We find that appending NEDD8 to the N terminus of the model Ag OVA resulted in degradation by both the proteasome and the autophagy protein degradation pathways, but only proteasomal degradation, involving the proteasomal subunit NEDD8 ultimate buster 1, resulted in peptide presentation. When directly compared with Ub, NEDD8 fusion was less efficient at generating peptides. However, inactivation of the NEDD8-conugation machinery by treating cells with MLN4924 inhibited the presentation of peptides from the defective ribosomal product-derived form of a model Ag. These results demonstrate that NEDD8 activity in the cell is important for direct Ag presentation, but not by directly targeting proteins for degradation., (Copyright © 2022 by The American Association of Immunologists, Inc.)- Published
- 2022
- Full Text
- View/download PDF
13. Correction: Influenza A Virus Negative Strand RNA Is Translated for CD8 + T Cell Immunosurveillance.
- Author
-
Hickman HD, Mays JW, Gibbs J, Kosik I, Magadán JG, Takeda K, Das S, Reynoso GV, Ngudiankama BF, Wei J, Shannon JP, McManus D, and Yewdell JW
- Published
- 2018
- Full Text
- View/download PDF
14. Influenza A Virus Negative Strand RNA Is Translated for CD8 + T Cell Immunosurveillance.
- Author
-
Hickman HD, Mays JW, Gibbs J, Kosik I, Magadán JG, Takeda K, Das S, Reynoso GV, Ngudiankama BF, Wei J, Shannon JP, McManus D, and Yewdell JW
- Subjects
- Animals, HeLa Cells, Humans, Mice, Mice, Inbred C57BL, Mice, Transgenic, Orthomyxoviridae Infections immunology, Protein Biosynthesis physiology, RNA, Viral genetics, CD8-Positive T-Lymphocytes immunology, Immunologic Surveillance immunology, Influenza A virus genetics, Influenza A virus immunology, RNA, Viral immunology
- Abstract
Probing the limits of CD8
+ T cell immunosurveillance, we inserted the SIINFEKL peptide into influenza A virus (IAV)-negative strand gene segments. Although IAV genomic RNA is considered noncoding, there is a conserved, relatively long open reading frame present in segment 8, encoding a potential protein termed NEG8. The biosynthesis of NEG8 from IAV has yet to be demonstrated. Although we failed to detect NEG8 protein expression in IAV-infected mouse cells, cell surface Kb -SIINFEKL complexes are generated when SIINFEKL is genetically appended to the predicted C terminus of NEG8, as shown by activation of OT-I T cells in vitro and in vivo. Moreover, recombinant IAV encoding of SIINFEKL embedded in the negative strand of the neuraminidase-stalk coding sequence also activates OT-I T cells in mice. Together, our findings demonstrate both the translation of sequences on the negative strand of a single-stranded RNA virus and its relevance in antiviral immunosurveillance.- Published
- 2018
- Full Text
- View/download PDF
15. Varied Role of Ubiquitylation in Generating MHC Class I Peptide Ligands.
- Author
-
Wei J, Zanker D, Di Carluccio AR, Smelkinson MG, Takeda K, Seedhom MO, Dersh D, Gibbs JS, Yang N, Jadhav A, Chen W, and Yewdell JW
- Subjects
- Animals, Cell Line, Cytosol chemistry, Cytosol immunology, Enzyme Inhibitors pharmacology, Histocompatibility Antigens Class I metabolism, Humans, Influenza A virus chemistry, Influenza A virus immunology, Kinetics, Ligands, Mice, Monitoring, Immunologic, Peptides metabolism, Pyrazoles, Pyrimidines, Sulfides, Ubiquitination, Vaccinia virus chemistry, Vaccinia virus immunology, Antigen Presentation, Histocompatibility Antigens Class I immunology, Nucleosides pharmacology, Peptides immunology, Sulfonamides pharmacology, T-Lymphocytes immunology
- Abstract
CD8
+ T cell immunosurveillance is based on recognizing oligopeptides presented by MHC class I molecules. Despite decades of study, the importance of protein ubiquitylation to peptide generation remains uncertain. In this study, we examined the ability of MLN7243, a recently described ubiquitin-activating enzyme E1 inhibitor, to block overall cytosolic peptide generation and generation of specific peptides from vaccinia- and influenza A virus-encoded proteins. We show that MLN7243 rapidly inhibits ubiquitylation in a variety of cell lines and can profoundly reduce the generation of cytosolic peptides. Kinetic analysis of specific peptide generation reveals that ubiquitylation of defective ribosomal products is rate limiting in generating class I peptide complexes. More generally, our findings demonstrate that the requirement for ubiquitylation in MHC class I-restricted Ag processing varies with class I allomorph, cell type, source protein, and peptide context. Thus, ubiquitin-dependent and -independent pathways robustly contribute to MHC class I-based immunosurveillance., (Copyright © 2017 by The American Association of Immunologists, Inc.)- Published
- 2017
- Full Text
- View/download PDF
16. Defining Viral Defective Ribosomal Products: Standard and Alternative Translation Initiation Events Generate a Common Peptide from Influenza A Virus M2 and M1 mRNAs.
- Author
-
Yang N, Gibbs JS, Hickman HD, Reynoso GV, Ghosh AK, Bennink JR, and Yewdell JW
- Subjects
- Animals, Antigen Presentation drug effects, Cell Line, Defective Viruses chemistry, Defective Viruses drug effects, Defective Viruses metabolism, Genes, MHC Class I, HeLa Cells, Humans, Influenza A virus chemistry, Influenza A virus drug effects, Influenza A virus metabolism, Mice, Mice, Inbred C57BL, Peptides chemistry, Protein Biosynthesis, Pyrans pharmacology, RNA Splicing drug effects, Spiro Compounds pharmacology, Defective Viruses genetics, Influenza A virus genetics, Peptides genetics, Ribosomes metabolism, Viral Matrix Proteins genetics
- Abstract
Influenza A virus gene segment 7 encodes two proteins: the M1 protein translated from unspliced mRNA and the M2 protein produced by mRNA splicing and largely encoded by the M1 +1 reading frame. To better understand the generation of defective ribosomal products relevant to MHC class I Ag presentation, we engineered influenza A virus gene segment 7 to encode the model H-2 K(b) class I peptide ligand SIINFEKL at the M2 protein C terminus. Remarkably, after treating virus-infected cells with the RNA splicing inhibitor spliceostatin A to prevent M2 mRNA generation, K(b)-SIINFEKL complexes were still presented on the cell surface at levels ≤60% of untreated cells. Three key findings indicate that SIINFEKL is produced by cytoplasmic translation of unspliced M1 mRNA initiating at CUG codons within the +1 reading frame: 1) synonymous mutation of CUG codons in the M2-reading frame reduced K(b)-SIINFEKL generation; 2) K(b)-SIINFEKL generation was not affected by drug-mediated inhibition of AUG-initiated M1 synthesis; and 3) K(b)-SIINFEKL was generated in vitro and in vivo from mRNA synthesized in the cytoplasm by vaccinia virus, and hence cannot be spliced. These findings define a viral defective ribosomal product generated by cytoplasmic noncanonical translation and demonstrate the participation of CUG-codon-based translation initiation in pathogen immunosurveillance., (Copyright © 2016 by The American Association of Immunologists, Inc.)
- Published
- 2016
- Full Text
- View/download PDF
17. Mixed proteasomes function to increase viral peptide diversity and broaden antiviral CD8+ T cell responses.
- Author
-
Zanker D, Waithman J, Yewdell JW, and Chen W
- Subjects
- Animals, CD8-Positive T-Lymphocytes enzymology, Coculture Techniques, Influenza A virus pathogenicity, Mice, Mice, Inbred C57BL, Mice, Knockout, Orthomyxoviridae Infections enzymology, Orthomyxoviridae Infections immunology, Orthomyxoviridae Infections prevention & control, Proteasome Endopeptidase Complex administration & dosage, Proteasome Endopeptidase Complex genetics, Tumor Cells, Cultured, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes virology, Influenza A virus immunology, Peptide Biosynthesis genetics, Peptide Biosynthesis immunology, Proteasome Endopeptidase Complex immunology, Up-Regulation immunology, Viral Proteins antagonists & inhibitors, Viral Proteins biosynthesis
- Abstract
The three proteasome subunits with proteolytic activity are encoded by standard or immunoproteasome genes. Many proteasomes expressed by normal cells and cells exposed to cytokines are "mixed", that is, contain both standard and immunoproteasome subunits. Using a panel of 38 defined influenza A virus-derived epitopes recognized by C57BL/6 mouse CD8(+) T cells, we used mice with targeted disruption of β1i, β2i, or β5i/β2i genes to examine the contribution of mixed proteasomes to the immunodominance hierarchy of antiviral CD8(+) T cells. We show that each immunoproteasome subunit has large effects on the primary and recall immunodominance hierarchies due to modulating both the available T cell repertoire and generation of individual epitopes as determined both biochemically and kinetically in Ag presentation assays. These findings indicate that mixed proteasomes function to enhance the diversity of peptides and support a broad CD8(+) T cell response.
- Published
- 2013
- Full Text
- View/download PDF
18. Distinct pathways generate peptides from defective ribosomal products for CD8+ T cell immunosurveillance.
- Author
-
Dolan BP, Li L, Veltri CA, Ireland CM, Bennink JR, and Yewdell JW
- Subjects
- Animals, Antigen Presentation drug effects, Antigen Presentation genetics, CD8-Positive T-Lymphocytes drug effects, CD8-Positive T-Lymphocytes metabolism, Cell Line, Tumor, Cell Membrane Permeability drug effects, Cell Membrane Permeability genetics, Cell Membrane Permeability immunology, Female, Green Fluorescent Proteins biosynthesis, Green Fluorescent Proteins genetics, H-2 Antigens biosynthesis, H-2 Antigens genetics, Mice, Mice, Inbred C57BL, Morpholines pharmacology, Ovalbumin immunology, Ovalbumin metabolism, Peptide Biosynthesis drug effects, Peptide Biosynthesis genetics, Peptide Fragments immunology, Peptide Fragments metabolism, Protein Stability drug effects, Recombinant Fusion Proteins biosynthesis, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Ribosomal Proteins genetics, Signal Transduction drug effects, Signal Transduction genetics, Antigen Presentation immunology, CD8-Positive T-Lymphocytes immunology, Immunologic Surveillance drug effects, Immunologic Surveillance genetics, Peptide Biosynthesis immunology, Ribosomal Proteins biosynthesis, Ribosomal Proteins deficiency, Signal Transduction immunology
- Abstract
To understand better the endogenous sources of MHC class I peptide ligands, we generated an antigenic reporter protein whose degradation is rapidly and reversibly controlled with Shield-1, a cell-permeant drug. Using this system, we demonstrate that defective ribosomal products (DRiPs) represent a major and highly efficient source of peptides and are completely resistant to our attempts to stabilize the protein. Although peptides also derive from nascent Shield-1-sensitive proteins and "retirees" created by Shield-1 withdrawal, these are much less efficient sources on a molar basis. We use this system to identify two drugs--each known to inhibit polyubiquitin chain disassembly--that selectively inhibit presentation of Shield-1-resistant DRiPs. These findings provide the initial evidence for distinct biochemical pathways for presentation of DRiPs versus retirees and implicate polyubiquitin chain disassembly or the actions of deubiquitylating enzymes as playing an important role in DRiP presentation.
- Published
- 2011
- Full Text
- View/download PDF
19. RNA polymerase II inhibitors dissociate antigenic peptide generation from normal viral protein synthesis: a role for nuclear translation in defective ribosomal product synthesis?
- Author
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Dolan BP, Knowlton JJ, David A, Bennink JR, and Yewdell JW
- Subjects
- Active Transport, Cell Nucleus drug effects, Active Transport, Cell Nucleus genetics, Active Transport, Cell Nucleus immunology, Animals, Antigen Presentation genetics, Antigen Presentation immunology, Antigens, Viral genetics, Antigens, Viral metabolism, Cell Line, Dogs, HeLa Cells, Humans, Influenza A virus drug effects, Influenza A virus enzymology, Influenza A virus immunology, L Cells, Mice, Neuraminidase antagonists & inhibitors, Neuraminidase biosynthesis, Neuraminidase genetics, Ovalbumin biosynthesis, Peptide Biosynthesis genetics, Peptide Fragments biosynthesis, Protein Biosynthesis drug effects, RNA, Messenger antagonists & inhibitors, RNA, Messenger biosynthesis, RNA, Messenger metabolism, Ribosomal Proteins biosynthesis, Ribosomal Proteins genetics, Viral Proteins genetics, Viral Proteins metabolism, Antigens, Viral biosynthesis, DNA-Directed RNA Polymerases antagonists & inhibitors, Dichlororibofuranosylbenzimidazole pharmacology, Peptide Biosynthesis drug effects, Peptide Biosynthesis immunology, Protein Biosynthesis immunology, Ribosomal Proteins deficiency, Viral Proteins biosynthesis
- Abstract
Following viral infection, cells rapidly present peptides from newly synthesized viral proteins on MHC class I molecules, likely from rapidly degraded forms of nascent proteins. The nature of these defective ribosomal products (DRiPs) remains largely undefined. Using inhibitors of RNA polymerase II that block influenza A virus neuraminidase (NA) mRNA export from the nucleus and inhibit cytoplasmic NA translation, we demonstrate a surprising disconnect between levels of NA translation and generation of SIINFEKL peptide genetically inserted into the NA stalk. A 33-fold reduction in NA expression is accompanied by only a 5-fold reduction in K(b)-SIINFEKL complex cell-surface expression, resulting in a net 6-fold increase in the overall efficiency of Ag presentation. Although the proteasome inhibitor MG132 completely blocked K(b)-SIINFEKL complex generation, we were unable to biochemically detect a MG132-dependent cohort of NA DRiPs relevant for Ag processing, suggesting that a minute population of DRiPs is a highly efficient source of antigenic peptides. These data support the idea that Ag processing uses compartmentalized translation, perhaps even in the nucleus itself, to increase the efficiency of the generation of class I peptide ligands.
- Published
- 2010
- Full Text
- View/download PDF
20. Unexpected role for the immunoproteasome subunit LMP2 in antiviral humoral and innate immune responses.
- Author
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Hensley SE, Zanker D, Dolan BP, David A, Hickman HD, Embry AC, Skon CN, Grebe KM, Griffin TA, Chen W, Bennink JR, and Yewdell JW
- Subjects
- Animals, Antibodies, Viral metabolism, Antigen Presentation genetics, Antigen Presentation immunology, B-Lymphocytes enzymology, B-Lymphocytes immunology, B-Lymphocytes virology, Cells, Cultured, Cysteine Endopeptidases deficiency, Cysteine Endopeptidases genetics, Dendritic Cells enzymology, Dendritic Cells immunology, Dendritic Cells virology, Mice, Mice, Congenic, Mice, Inbred C57BL, Mice, Knockout, NF-kappa B antagonists & inhibitors, NF-kappa B physiology, Proteasome Endopeptidase Complex deficiency, Proteasome Endopeptidase Complex genetics, Protein Subunits deficiency, Protein Subunits genetics, Signal Transduction genetics, Signal Transduction immunology, Antibodies, Viral biosynthesis, Cysteine Endopeptidases physiology, Immunity, Innate genetics, Influenza A virus immunology, Proteasome Endopeptidase Complex physiology, Protein Subunits physiology
- Abstract
Proteasomes are multisubunit proteases that initiate degradation of many Ags presented by MHC class I molecules. Vertebrates express alternate forms of each of the three catalytic proteasome subunits: standard subunits, and immunosubunits, which are constitutively expressed by APCs and are induced in other cell types by exposure to cytokines. The assembly of mixed proteasomes containing standard subunits and immunosubunits is regulated in a tissue specific manner. In this study, we report that the presence of mixed proteasomes in immune cells in LMP2(-/-) mice compromises multiple components that contribute to the generation of antiviral Ab responses, including splenic B cell numbers, survival and function of adoptively transferred B cells, Th cell function, and dendritic cell secretion of IL-6, TNF-alpha, IL-1beta, and type I IFNs. These defects did not result from compromised overall protein degradation, rather they were associated with altered NF-kappaB activity. These findings demonstrate an important role for immunoproteasomes in immune cell function beyond their contribution to Ag processing.
- Published
- 2010
- Full Text
- View/download PDF
21. Defective ribosomal products are the major source of antigenic peptides endogenously generated from influenza A virus neuraminidase.
- Author
-
Dolan BP, Li L, Takeda K, Bennink JR, and Yewdell JW
- Subjects
- Amino Acid Sequence, Animals, Antigens, Viral metabolism, Cell Line, Dendritic Cells enzymology, Dendritic Cells immunology, Dendritic Cells virology, Dogs, Enzyme Activation immunology, Enzyme Stability immunology, Epitopes biosynthesis, Epitopes metabolism, Fibroblasts enzymology, Fibroblasts immunology, Fibroblasts virology, H-2 Antigens biosynthesis, H-2 Antigens metabolism, L Cells, Mice, Molecular Sequence Data, Monocytes enzymology, Monocytes immunology, Monocytes virology, Neuraminidase biosynthesis, Orthomyxoviridae Infections enzymology, Orthomyxoviridae Infections immunology, Orthomyxoviridae Infections virology, Ovalbumin metabolism, Ovalbumin physiology, Peptide Fragments metabolism, Peptide Fragments physiology, Protein Folding, Protein Transport immunology, Ribosomal Proteins metabolism, Antigen Presentation immunology, Antigens, Viral biosynthesis, Influenza A Virus, H1N1 Subtype enzymology, Influenza A Virus, H1N1 Subtype immunology, Neuraminidase metabolism, Protein Biosynthesis immunology, Ribosomal Proteins biosynthesis, Ribosomal Proteins deficiency
- Abstract
The defective ribosomal product (DRiP) hypothesis of endogenous Ag processing posits that rapidly degraded forms of nascent proteins are a major source of peptide ligands for MHC class I molecules. Although there is broad experimental support for the DRiP hypothesis, careful kinetic analysis of the generation of defined peptide class I complexes has been limited to studies of recombinant vaccinia viruses expressing genes derived from other organisms. In this study, we show that insertion of the SIINFEKL peptide into the stalk of influenza A virus neuraminidase (NA) does not detectably modify NA folding, degradation, transport, or sp. act. when expressed in its natural context of influenza A virus infection. Using the 25-D1.16 mAb specific for K(b)-SIINFEKL to precisely quantitate cell surface complexes by flow cytometry, we demonstrate that SIINFEKL is generated in complete lockstep with initiation and abrogation of NA biosynthesis in both L-K(b) fibroblast cells and DC2.4 dendritic/monocyte cells. SIINFEKL presentation requires active proteasomes and TAP, consistent with its generation from a cytosolic DRiP pool. From the difference in the shutoff kinetics of K(b)-SIINFEKL complex expression following protein synthesis versus proteasome inhibition, we estimate that the t(1/2) of the biosynthetic source of NA peptide is approximately 5 min. These observations extend the relevance of the DRiP hypothesis to viral proteins generated in their natural context.
- Published
- 2010
- Full Text
- View/download PDF
22. Cutting edge: Sympathetic nervous system increases proinflammatory cytokines and exacerbates influenza A virus pathogenesis.
- Author
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Grebe KM, Takeda K, Hickman HD, Bailey AL, Embry AC, Bennink JR, and Yewdell JW
- Subjects
- Animals, Cell Movement immunology, Immunity, Innate, Interleukin-1 biosynthesis, Mice, Orthomyxoviridae Infections immunology, Pneumonia virology, Survival Rate, Sympathectomy, Cytokines biosynthesis, Inflammation immunology, Influenza A virus pathogenicity, Sympathetic Nervous System immunology
- Abstract
Although the sympathetic nervous system innervates the lung, little is known about its participation in host immunity to pulmonary pathogens. In this study, we show that peripheral sympathectomy reduces mouse morbidity and mortality from influenza A virus-induced pneumonia due to reduced inflammatory influx of monocytes, neutrophils, and NK cells. Mortality was also delayed by treating mice with an alpha-adrenergic antagonist. Sympathectomy diminished the immediate innate cytokine responses, particularly IL-1, which was profoundly reduced. These findings demonstrate an unexpected role for the sympathetic nervous system in innate antiviral immunity and in exacerbating the pathology of a virus of great significance to human and animal health.
- Published
- 2010
- Full Text
- View/download PDF
23. Efficient cross-priming of antiviral CD8+ T cells by antigen donor cells is GRP94 independent.
- Author
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Lev A, Dimberu P, Das SR, Maynard JC, Nicchitta CV, Bennink JR, and Yewdell JW
- Subjects
- Animals, Antiviral Agents metabolism, CD8-Positive T-Lymphocytes transplantation, Cell Line, Female, Gene Knockdown Techniques, H-2 Antigens genetics, H-2 Antigens immunology, H-2 Antigens metabolism, Humans, Influenza A virus genetics, Influenza A virus immunology, Membrane Glycoproteins biosynthesis, Membrane Glycoproteins deficiency, Membrane Glycoproteins genetics, Mice, Mice, Inbred C57BL, Mice, Transgenic, Peptide Fragments genetics, Peptide Fragments immunology, Peptide Fragments metabolism, RNA, Small Interfering genetics, Vaccinia virus genetics, Vaccinia virus immunology, Vesicular stomatitis Indiana virus genetics, Vesicular stomatitis Indiana virus immunology, Antigens, Viral administration & dosage, Antigens, Viral immunology, Antiviral Agents immunology, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes virology, Cross-Priming immunology, Membrane Glycoproteins physiology
- Abstract
Cross-priming, the activation of naive CD8+ T cells by dendritic cells presenting Ags synthesized by other cells, is believed to play an important role in the generation of antiviral and antitumor responses. The molecular mechanism(s) underlying cross-priming remain poorly defined and highly controversial. GRP94 (gp96), an abundant endoplasmic reticulum chaperone with innate immune-activating capacity, has been widely reported to play a major role in cross-priming. In this study, we show that cells whose expression of GRP94 is silenced via transient or stable transfection with GRP94-directed small interfering RNAs demonstrate no reduction in their abilities to generate class I peptide complexes in cultured cells or to prime antiviral CD8+ T cell responses in vivo. In demonstrating the dispensability of GRP94, our finding points to the importance of alternative mechanisms for generation of class I peptide complexes from endogenous and exogenous Ags and immunogens.
- Published
- 2009
- Full Text
- View/download PDF
24. Quantitating T cell cross-reactivity for unrelated peptide antigens.
- Author
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Ishizuka J, Grebe K, Shenderov E, Peters B, Chen Q, Peng Y, Wang L, Dong T, Pasquetto V, Oseroff C, Sidney J, Hickman H, Cerundolo V, Sette A, Bennink JR, McMichael A, and Yewdell JW
- Subjects
- Animals, CD8-Positive T-Lymphocytes microbiology, CD8-Positive T-Lymphocytes virology, Cell Line, Tumor, Clone Cells, Cross Reactions, Cross-Priming immunology, Humans, Male, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Knockout, Mice, Transgenic, Peptide Fragments agonists, Peptide Library, Predictive Value of Tests, Protein Binding immunology, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes metabolism, Epitopes, T-Lymphocyte immunology, Epitopes, T-Lymphocyte metabolism, Peptide Fragments immunology, Peptide Fragments metabolism
- Abstract
Quantitating the frequency of T cell cross-reactivity to unrelated peptides is essential to understanding T cell responses in infectious and autoimmune diseases. Here we used 15 mouse or human CD8+ T cell clones (11 antiviral, 4 anti-self) in conjunction with a large library of defined synthetic peptides to examine nearly 30,000 TCR-peptide MHC class I interactions for cross-reactions. We identified a single cross-reaction consisting of an anti-self TCR recognizing a poxvirus peptide at relatively low sensitivity. We failed to identify any cross-reactions between the synthetic peptides in the panel and polyclonal CD8+ T cells raised to viral or alloantigens. These findings provide the best estimate to date of the frequency of T cell cross-reactivity to unrelated peptides ( approximately 1/30,000), explaining why cross-reactions between unrelated pathogens are infrequently encountered and providing a critical parameter for understanding the scope of self-tolerance.
- Published
- 2009
- Full Text
- View/download PDF
25. Terminal deoxynucleotidyl transferase establishes and broadens antiviral CD8+ T cell immunodominance hierarchies.
- Author
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Haeryfar SM, Hickman HD, Irvine KR, Tscharke DC, Bennink JR, and Yewdell JW
- Subjects
- Animals, Antigen Presentation immunology, Antigens, Viral immunology, DNA Nucleotidylexotransferase deficiency, DNA Nucleotidylexotransferase genetics, Female, Influenza A virus immunology, Male, Mice, Mice, Inbred BALB C, Mice, Knockout, Receptors, Antigen, T-Cell immunology, Vaccinia virus immunology, CD8-Positive T-Lymphocytes enzymology, CD8-Positive T-Lymphocytes immunology, DNA Nucleotidylexotransferase immunology, DNA Nucleotidylexotransferase metabolism
- Abstract
The action of TdT on mouse TCR genes accounts for approximately 90% of T cell repertoire diversity. We report that in TdT-/- mice, total T(CD8+) responses to influenza and vaccinia viruses are reduced by approximately 30% relative to wild-type mice. We find that T(CD8+) responses to three subdominant influenza virus determinants are reduced to background values in TdT-/- mice while responses to three immunodominant determinants undergo a major reshuffling. A similar reshuffling occurs in T(CD8+) responses to immunodominant vaccinia virus determinants, and is clearly based on broad differences in TCR family usage and CDR3 length between wild-type and TdT-/- mice. These findings demonstrate that TdT plays a critical role in the magnitude and breadth of anti-viral T(CD8+) responses toward individual determinants and suggests that germline TCR repertoire bias toward the most dominant determinants is a major factor in establishing immunodominance hierarchies.
- Published
- 2008
- Full Text
- View/download PDF
26. Cutting edge: MHC class I-Ly49 interaction regulates neuronal function.
- Author
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Zohar O, Reiter Y, Bennink JR, Lev A, Cavallaro S, Paratore S, Pick CG, Brooker G, and Yewdell JW
- Subjects
- Animals, Cell Line, Gene Expression, Immunohistochemistry, In Situ Hybridization, Mice, Neurons cytology, RNA, Messenger analysis, Receptors, NK Cell Lectin-Like, Signal Transduction physiology, Synapses metabolism, Antigens, Ly metabolism, Cerebral Cortex growth & development, Cerebral Cortex metabolism, Histocompatibility Antigens Class I metabolism, Lectins, C-Type metabolism, Neurons metabolism
- Abstract
MHC class I molecules (MHC-I) have been implicated in nervous system development in the mouse. In this study we present evidence for the interaction of MHC-I with the NK cell receptor Ly49 in primary cortical neuronal cultures. We show that MHC-I and Ly49 are expressed on neuronal soma and axon surfaces, with Ly49 also present on dendrites. Anti-MHC-I Abs reduce synapsin-I expression and enhance neurite outgrowth and neuronal death. Conversely, anti-Ly49 mAbs increase synapsin-I expression, reduce neurite outgrowth, and promote neuron viability. Because we show that Ly49 genes are selectively expressed in the adult brain, these findings suggest an unsuspected role for the MHC-I-Ly49 interaction in the development and function of the brain.
- Published
- 2008
- Full Text
- View/download PDF
27. CD8 alpha alpha-mediated intraepithelial lymphocyte snatching of thymic leukemia MHC class Ib molecules in vitro and in vivo.
- Author
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Pardigon N, Takeda K, Saunier B, Hornung F, Gibbs J, Weisberg A, Contractor N, Kelsall B, Bennink JR, and Yewdell JW
- Subjects
- Animals, Biological Transport, Active genetics, Biological Transport, Active immunology, CD8 Antigens biosynthesis, CD8 Antigens genetics, Cell Communication genetics, Cell Communication immunology, Cell Line, Tumor, Cell Membrane genetics, Cell Membrane immunology, Cell Membrane metabolism, Coculture Techniques, Epithelial Cells metabolism, Intestinal Mucosa cytology, Intestinal Mucosa metabolism, Lymphocyte Subsets metabolism, Membrane Glycoproteins biosynthesis, Membrane Glycoproteins genetics, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Knockout, Signal Transduction genetics, Signal Transduction immunology, Transfection, Antigens, Neoplasm metabolism, CD8 Antigens physiology, Epithelial Cells immunology, H-2 Antigens metabolism, Intestinal Mucosa immunology, Lymphocyte Subsets immunology, Membrane Glycoproteins metabolism
- Abstract
Thymic leukemia (TL) is a MHC class Ib molecule that interacts with CD8alphaalpha homodimers. CD8alphaalpha is abundantly expressed by intraepithelial T lymphocytes (IELs) located in close proximity to TL-expressing intestinal epithelial cells. In this study, we show that CD8alphaalpha(+) IELs "snatch" TL from the plasma membrane of TL-expressing cells and express TL in its proper orientation on their own cell surface. TL snatching is enhanced by cross-linking of IEL TCRs in a phosphatidylinositol kinase-dependent manner, and results in overall alterations to the IEL cell surface detected by enhanced binding of peanut agglutinin lectin. Induction of bowel inflammation results in the presence of TL on IELs, probably via in vivo snatching, providing the initial evidence for the interaction of CD8alphaalpha IELs with intestinal cells.
- Published
- 2006
- Full Text
- View/download PDF
28. Tight linkage between translation and MHC class I peptide ligand generation implies specialized antigen processing for defective ribosomal products.
- Author
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Qian SB, Reits E, Neefjes J, Deslich JM, Bennink JR, and Yewdell JW
- Subjects
- Animals, Antigen Presentation drug effects, Antigens, Ly biosynthesis, Antigens, Ly metabolism, Cell Line, Tumor, Cycloheximide pharmacology, Egg Proteins biosynthesis, Egg Proteins metabolism, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, H-2 Antigens biosynthesis, L Cells, Ligands, Membrane Proteins biosynthesis, Membrane Proteins metabolism, Mice, Nucleoproteins genetics, Nucleoproteins metabolism, Ovalbumin biosynthesis, Ovalbumin metabolism, Peptide Biosynthesis drug effects, Peptide Biosynthesis genetics, Peptide Fragments, Peptides antagonists & inhibitors, Peptides immunology, Proteasome Endopeptidase Complex biosynthesis, Proteasome Endopeptidase Complex genetics, Proteasome Endopeptidase Complex metabolism, Protein Synthesis Inhibitors pharmacology, Protein Transport genetics, Protein Transport immunology, Recombinant Proteins genetics, Recombinant Proteins metabolism, Ribosomal Proteins antagonists & inhibitors, Ribosomal Proteins biosynthesis, Ribosomal Proteins genetics, Antigen Presentation genetics, H-2 Antigens genetics, H-2 Antigens metabolism, Peptide Biosynthesis immunology, Peptides genetics, Peptides metabolism, Protein Biosynthesis, Ribosomal Proteins metabolism
- Abstract
There is mounting evidence that MHC class I peptide ligands are predominantly generated from defective ribosomal products and other classes of polypeptides degraded rapidly (t1/2 < 10 min) following their synthesis. The most direct evidence supporting this conclusion is the rapid inhibition of peptide ligand generation following cycloheximide-mediated inhibition of protein synthesis. In this study, we show that this linkage is due to depleting the pool of rapidly degraded proteins, and not to interference with other protein synthesis-dependent processes. Our findings indicate that in the model systems used in this study, MHC class I peptides are preferentially generated from rapidly degraded polypeptides relative to slowly degraded proteins. This conclusion is supported by the properties of peptide presentation from slowly degraded (t1/2 = 4 h) defective ribosomal products generated artificially by incorporation of the amino acid analog canavanine into a model viral Ag. We propose that specialized machinery exists to link protein synthesis with class I peptide ligand generation to enable the rapid detection of viral gene expression.
- Published
- 2006
- Full Text
- View/download PDF
29. HLA-A*0201, HLA-A*1101, and HLA-B*0702 transgenic mice recognize numerous poxvirus determinants from a wide variety of viral gene products.
- Author
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Pasquetto V, Bui HH, Giannino R, Banh C, Mirza F, Sidney J, Oseroff C, Tscharke DC, Irvine K, Bennink JR, Peters B, Southwood S, Cerundolo V, Grey H, Yewdell JW, and Sette A
- Subjects
- Animals, Antigens, Viral genetics, Antigens, Viral immunology, Epitopes, T-Lymphocyte genetics, HLA-A Antigens immunology, HLA-A11 Antigen, HLA-A2 Antigen, HLA-B Antigens immunology, HLA-B7 Antigen, Humans, Mice, Mice, Inbred BALB C, Mice, Transgenic, Predictive Value of Tests, Vaccinia virus genetics, Viral Proteins genetics, Viral Proteins immunology, Epitopes, T-Lymphocyte immunology, HLA-A Antigens genetics, HLA-B Antigens genetics, Vaccinia virus immunology
- Abstract
In virus models explored in detail in mice, CTL typically focus on a few immunodominant determinants. In this study we use a multipronged approach to understand the diversity of CTL responses to vaccinia virus, a prototypic poxvirus with a genome approximately 20-fold larger than that of the model RNA viruses typically studied in mice. Based on predictive computational algorithms for peptide binding to HLA supertypes, we synthesized a panel of 2889 peptides to begin to create an immunomic map of human CTL responses to poxviruses. Using this panel in conjunction with CTLs from vaccinia virus-infected HLA transgenic mice, we identified 14 HLA-A*0201-, 4 HLA-A*1101-, and 3 HLA-B*0702-restricted CD8(+) T cell determinants distributed over 20 distinct proteins. These peptides were capable of binding one or multiple A2, A3, and B7 supertype molecules with affinities typical of viral determinants. Surprisingly, many of the viral proteins recognized are predicted to be late gene products, in addition to the early intermediate gene products expected. Nearly all of the determinants identified have identical counterparts encoded by modified vaccinia virus Ankara as well as variola virus, the agent of smallpox. These findings have implications for the design of new smallpox vaccines and the understanding of immune responses to large DNA viruses in general.
- Published
- 2005
- Full Text
- View/download PDF
30. Regulatory T cells suppress CD8+ T cell responses induced by direct priming and cross-priming and moderate immunodominance disparities.
- Author
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Haeryfar SM, DiPaolo RJ, Tscharke DC, Bennink JR, and Yewdell JW
- Subjects
- Amino Acid Sequence, Animals, Antigens, Polyomavirus Transforming genetics, Antigens, Viral genetics, CD4 Antigens genetics, CD4 Antigens metabolism, Cytotoxicity, Immunologic, Female, Immune Tolerance, Immunodominant Epitopes genetics, Influenza A virus genetics, Influenza A virus immunology, Lymphocyte Depletion, Mice, Mice, Inbred C57BL, Mice, Knockout, Peptide Fragments genetics, Peptide Fragments immunology, Vaccinia virus genetics, Vaccinia virus immunology, CD4-Positive T-Lymphocytes immunology, T-Lymphocytes immunology
- Abstract
Little is known regarding the participation of CD4+ CD25+ regulatory T cells (Treg) in TCD8+ responses. In this study, we show that Treg depletion via treatment with anti-CD25 mAb (PC61) significantly enhances TCD8+ responses to influenza A virus, vaccinia virus, and SV40-transformed cells induced by either direct priming or cross-priming. PC61 did not enhance TCD8+ responses in CD4-deficient mice, providing the initial demonstration that PC61 acts on a subset of TCD4+, and not on other cells that express either CD25 or a fortuitously cross-reactive Ag. We further show that Treg selectively suppress responses to the most immunodominant TCD8+ determinants in the three systems examined. Therefore, Treg influence TCD8 immunodominance hierarchies by moderating disparities in responses to different determinants.
- Published
- 2005
- Full Text
- View/download PDF
31. Reversal in the immunodominance hierarchy in secondary CD8+ T cell responses to influenza A virus: roles for cross-presentation and lysis-independent immunodomination.
- Author
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Chen W, Pang K, Masterman KA, Kennedy G, Basta S, Dimopoulos N, Hornung F, Smyth M, Bennink JR, and Yewdell JW
- Subjects
- Adoptive Transfer, Animals, Cell Line, Tumor, Immunologic Memory, Mice, Mice, Inbred C57BL, CD8-Positive T-Lymphocytes immunology, Cross-Priming, Cytotoxicity, Immunologic, Influenza A virus immunology
- Abstract
Immunodominance is a central feature of CD8+ T cell (TCD8+) responses to pathogens, transplants, and tumors. Determinants occupy a stable position in an immunodominance hierarchy (alpha-, beta-, etc.) defined by the frequencies of responding TCD8+. In this paper, we study the mechanistic basis for place-swapping between alpha- (acid polymerase (PA)(224-233)) and beta-determinants (nuclear protein 366-374) in primary vs secondary anti-influenza A virus (IAV) responses in mice. This phenomena was recently correlated with the inability of IAV-infected nondendritic cells (DCs) to generate PA(224-233), and it was proposed that secondary TCD8+ are principally activated by IAV-infected epithelial cells, while primary TCD8+ are activated by IAV-infected DCs. In this study, we show that the inability of non-DCs to generate PA(224-232) is relative rather than absolute, and that the preferential use of cross-priming in secondary anti-IAV responses can also account for the revised hierarchy. We further show that immunodomination of PA(224-233)-specific TCD8+ by nucleoprotein 366-374-specific TCD8+ plays a critical role in the phenomena, and that this is unlikely to be mediated by TCD8+ lysis of APCs or other cells.
- Published
- 2004
- Full Text
- View/download PDF
32. Inhibitory effects of cytomegalovirus proteins US2 and US11 point to contributions from direct priming and cross-priming in induction of vaccinia virus-specific CD8(+) T cells.
- Author
-
Basta S, Chen W, Bennink JR, and Yewdell JW
- Subjects
- Animals, CD4-Positive T-Lymphocytes physiology, Female, H-2 Antigens analysis, H-2 Antigens physiology, Histocompatibility Antigen H-2D, Interferon-gamma biosynthesis, Lymphocyte Activation, Mice, Mice, Inbred C57BL, Tumor Necrosis Factor-alpha biosynthesis, CD8-Positive T-Lymphocytes immunology, RNA-Binding Proteins physiology, Vaccinia virus immunology, Viral Envelope Proteins physiology, Viral Proteins physiology
- Abstract
The extent to which naive CD8(+) CTLs (T(CD8)(+)) are primed by APCs presenting endogenous Ags (direct priming) or Ags acquired from other infected cells (cross-priming) is a critical topic in basic and applied immunology. To examine the contribution of direct priming in the induction of VV-specific T(CD8)(+), we generated recombinant vaccinia viruses that express human CMV proteins (US2 and US11) that induce the destruction of newly synthesized MHC class I molecules. Expression of US2 or US11 was associated with a 24-63% decrease in numbers of primary or secondary VV-specific T(CD8)(+) responding to i.p. infection. Using HPLC-isolated peptides from VV-infected cells, we show that US2 and US11 selectively inhibit T(CD8)(+) responses to a subset of immunogenic VV determinants. Moreover, VV-US2 and lysates from VV-infected histoincompatible cells elicit T(CD8)(+) specific for a similar subset of VV determinants. These findings indicate that US2 and US11 can function in vivo to interfere with the activation of virus-specific T(CD8)(+). Furthermore, they suggest that 1) both cross-priming and direct priming contribute significantly to the generation of VV-specific T(CD8)(+), 2) the sets of immunogenic vaccinia virus determinants generated by cross-priming and direct priming are not completely overlapping, and 3) cross-priming overrides the effects of cis-acting viral interference with the class I Ag presentation pathway.
- Published
- 2002
- Full Text
- View/download PDF
33. Recycling CD1d1 molecules present endogenous antigens processed in an endocytic compartment to NKT cells.
- Author
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Roberts TJ, Sriram V, Spence PM, Gui M, Hayakawa K, Bacik I, Bennink JR, Yewdell JW, and Brutkiewicz RR
- Subjects
- Animals, Antigens, CD1d, Cell Line, Glycosylphosphatidylinositols physiology, Killer Cells, Natural immunology, Mice, Primaquine pharmacology, Rats, Antigen Presentation, Antigens, CD1 physiology, Endocytosis, Killer Cells, Natural metabolism
- Abstract
Mouse CD1d1 molecules present endogenous glycolipids to NKT cells. Although glycolipid presentation requires CD1d1 transport through the endocytic pathway, the processing requirements for such endogenous Ag presentation by CD1d1 molecules are undefined. We examined CD1d1 Ag presentation to NKT cells by disrupting endocytic trafficking and function in cells expressing normal and mutated CD1d1 expressed by recombinant vaccinia viruses. Consistent with previous studies, we found that preventing CD1d1 localization to endosomes by altering its cytoplasmic targeting sequences abrogated recognition by Valpha14Jalpha281(+) NKT cells without affecting recognition by Valpha14(-) NKT cells. Increasing the pH of acidic compartments by incubating cells with chloroquine or bafilomycin A1 blocked CD1d1 recognition by Valpha14(+) (but not Valpha14(-)) NKT cells without reducing levels of cell surface CD1d1. Similar results were obtained with primaquine, which interferes with the recycling of cell surface glycoproteins. These results suggest that the loading of a subset of glycolipid ligands onto CD1d1 molecules entails the delivery of cell surface CD1d1 molecules and an acidic environment in the endocytic pathway.
- Published
- 2002
- Full Text
- View/download PDF
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