Progress 11/01/07 to 10/31/12
Outputs
OUTPUTS: NON-TECHNICAL SUMMARY: VIruses and cancers are normally prevented or eliminated by the immune system. Nevertheless, both viral diseases and tumors are common problems in animals and man. In many cases this reflects adaptation of the pathogen or tumor to the immune system. The goal of this project is, first, to identify features of the immune system, and of pathogens, that allow pathogens to evade immune responses; and second, to identify ways of altering the immune response or the pathogen's life cycle in order to enhance the immune response and reduce the virulence of the pathogen. OBJECTIVES: Virus infections and cancerous cells are normally recognized and eliminated by the immune system. Nevertheless, viruses and tumors remain common problems in man and other animals. The goal of this project is to understand (1) the mechanisms by which the immune system normally identifies abnormal cells, (2) why this recognition sometimes fails, (3) how viruses evade recognition by the immune system, and (3) how to improve immune recognition in order to protect against, or eliminate, viruses and cancers. APPROACH: Previous and ongoing research has shown that ER-localized aminopeptidases help determine which epitopes in a virus are immunodominant. I will use cell culture models and knockout mice to determine which aminopeptidases are most important, and which subsets of epitopes are most dependent on individual aminopeptidases. I will then test the effect of altering levels of these aminopeptidases on antiviral immune responses, and on viral pathogenesis. As well I will identify viral molecules that inhibit immune recognition. I will test the effect of these molecules in cell culture and in animals, with the goal of enhancing antiviral immunity and preventive vaccines. KEYWORDS: immunity; antigen presentation; immune evasion; vaccine; virus; tumor; PARTICIPANTS: PARTICIPANTS: Individuals: Ian York: Principle investigator Barbara Christian: Research technician Training/Professional development: Jerome Moulden: Graduate student Ram Kamal: Graduate student. Collaborators: Roger Maes (DCPAH, MSU) Henry Hunt (ADOL, MSU) TARGET AUDIENCES: EFFORTS include mentoring two full-time Ph.D. students (Jerome Moulden and Ram Kamal), as well as one undergraduate student Eric Collins (as a Professorial Assistant, Spring '10). TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: This project is ending due to the departure of Ian York from MSU in Summer 2010.
Impacts
PROGRESS: 2010/01 TO 2010/12 OUTPUTS: ACTIVITIES in this project include: (1) Construction of allelic variants of the ER aminopeptidase ERAP1 that are associated with increased and reduced risk of ankylosing spondylitis; (2) Analysis of the effect of ERAP1 variants on MHC class I peptide presentation; (3) Construction of a mutant feline herpesvirus 1 bacterial artififial chromosome and of a mutant FHV-1 virus. lacking the immune evasion gene UL49.5; (4) Identification of the MHC class I immune evasion molecule of MDV, and of its mechanism of action; (5) Beginning construction of a mutant MDV lacking its immune evasion gene. (3) I am mentoring two full-time Ph.D. students (Jerome Moulden and Ram Kamal), as well as two undergraduate students, Cari Hearn (in a CVM Summer Research Program, Summer) and Eric Collins (as a Professorial Assistant, Spring '10). (4) Services include teaching classes for MMG813 (Graduate Virology), MMG101 (Preview of Microbiology), MMG103 (Frontiers of Microbiology and Molecular Genetics), MMG851 (Graduate Immunology), and MMG559 (Veterinary Microbiology & Immunology) IMPACT: 2010/01 TO 2010/12 1) We have identified effects of ERAP1 allelic variants on peptides associated with ankylosing spondylitis. (2) We have constructed a knockout FHV-1 lacking its immune evasion gene in order to understand the function of this gene in viral pathogenesis and immunity to viruses. We will test the efficacy of this knockout virus as a backbone for vaccines against FHV-1, predicting that reducing immune evasion will result in a highly antigenic, attenuated virus. (3) We have identified an MHC class I immune evasion gene of Marek's Disease Virus (MDV) of chickens and have clonied and expressed this gene in cultured cells. We are constructing a knockout virus lacking this gene in order to understand the function of this gene in viral pathogenesis and immunity to viruses. We will test the efficacy of this knockout virus as a backbone for vaccines against MDV, predicting that reducing immune evasion will result in a highly antigenic, attenuated virus.
Publications
- 1. Characterizing the Specificity and Co-operation of Aminopeptidases in the Cytosol and ER During MHC Class I antigen Presentation. (2010) Arron Hearn, Ian A. York, Courtney Bishop, and Kenneth L. Rock (Submitted, In revision) 2. An MHC class I immune evasion gene of Marek's Disease Virus. (2010) Cari Hearn, Likit Preeyanon, Henry D. Hunt, Ian A. York (In preparation) 3. Herpesvirus UL49.5 function and evolution as an immune evasion molecule. (2010) Jerome Moulden III, Roger Maes, Sheldon Tsai, Cari Hearn, and Ian York (In preparation)
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: OUTPUTS: ACTIVITIES in this project include: (1) Construction of allelic variants of the ER aminopeptidase ERAP1 that are associated with increased and reduced risk of ankylosing spondylitis; (2) Analysis of the effect of ERAP1 variants on MHC class I peptide presentation; (3) Construction of a mutant feline herpesvirus 1 bacterial artififial chromosome and of a mutant FHV-1 virus. lacking the immune evasion gene UL49.5; (4) Identification of the MHC class I immune evasion molecule of MDV, and of its mechanism of action; (5) Beginning construction of a mutant MDV lacking its immune evasion gene. (3) I am mentoring two full-time Ph.D. students (Jerome Moulden and Ram Kamal), as well as two undergraduate students, Cari Hearn (in a CVM Summer Research Program, Summer '09) and Eric Collins (as a Professorial Assistant, Fall '09 - Spring '10). (4) Events include students Ram Kamal and Jerome Moulden attending the Chicago Autumn Immunology Conference (Nov '09). (5) Services include teaching classes for MMG813 (Graduate Virology), MMG101 (Preview of Microbiology), MMG103 (Frontiers of Microbiology and Molecular Genetics), MMG851 (Graduate Immunology), and MMG559 (Veterinary Microbiology & Immunology) PARTICIPANTS: PARTICIPANTS: Individuals: Ian York: Principle investigator Barbara Christian: Research technician Training/Professional development: Jerome Moulden: Graduate student Ram Kamal: Graduate student. Collaborators: Roger Maes (DCPAH, MSU) Henry Hunt (ADOL, MSU) TARGET AUDIENCES: EFFORTS include mentoring two full-time Ph.D. students (Jerome Moulden and Ram Kamal), as well as two undergraduate students, Cari Hearn (in a CVM Summer Research Program, Summer '09) and Eric Collins (as a Professorial Assistant, Fall '09 - Spring '10). I have also taught classes for MMG813 (Graduate Virology, Spring '09), MMG101 (Preview of Microbiology (Fall '09)), MMG103 (Frontiers of Microbiology and Molecular Genetics (Spring '09)), MMG851 (Graduate Immunology (Fall '09)), and MMG559 (Veterinary Microbiology & Immunology (Fall '09)) TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
IMPACTS (1) We have identified effects of ERAP1 allelic variants on peptides associated with ankylosing spondylitis. (2) We have constructed a knockout FHV-1 lacking its immune evasion gene in order to understand the function of this gene in viral pathogenesis and immunity to viruses. We will test the efficacy of this knockout virus as a backbone for vaccines against FHV-1, predicting that reducing immune evasion will result in a highly antigenic, attenuated virus. (3) We have identified an MHC class I immune evasion gene of Marek's Disease Virus (MDV) of chickens and have clonied and expressed this gene in cultured cells. We are constructing a knockout virus lacking this gene in order to understand the function of this gene in viral pathogenesis and immunity to viruses. We will test the efficacy of this knockout virus as a backbone for vaccines against MDV, predicting that reducing immune evasion will result in a highly antigenic, attenuated virus.
Publications
- Kawahara, M., I.A. York, A. Hearn, D. Farfan, and K.L. Rock. (2009) Analysis of the role of tripeptidyl peptidase II in MHC Class I antigen presentation in vivo. J Immunol 183:6069-6077.
- Characterizing the Specificity and Co-operation of Aminopeptidases in the Cytosol and ER During MHC Class I antigen Presentation. (2010) Arron Hearn, Ian A. York, Courtney Bishop, and Kenneth L. Rock (Submitted, In revision)
- An MHC class I immune evasion gene of Marek's Disease Virus. (2010) Cari Hearn, Likit Preeyanon, Henry D. Hunt, Ian A. York (In preparation)
- Herpesvirus UL49.5 function and evolution as an immune evasion molecule. (2010) Jerome Moulden III, Roger Maes, Sheldon Tsai, Cari Hearn, and Ian York (In preparation)
- Dang, Y., X. Wang, T. Zhou, I.A. York, and Y.H. Zheng. (2009) Identification of a novel WxSLVK motif in the N terminus of human immunodeficiency virus and simian immunodeficiency virus Vif that is critical for APOBEC3G and APOBEC3F neutralization. J Virol 83:8544-8552
- Hearn, A., I.A. York, and K.L. Rock. (2009) The specificity of trimming of MHC Class I-presented peptides in the endoplasmic reticulum. J Immunol 183:5526-5536.
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: ACTIVITIES in this project include: (1) Analysis of peptide substrates for ER-localized aminopeptides and impact of aminopeptidases on antigen presentation. We are extending our research to assess the effect of aminopeptidase allelic variants on susceptibility to autoimmune disease, specifically ankylosing spondylitis, as a model for immunodominance in antigen processing and presentation. (2) Analysis of immune evasion molecules in feline herpesvirus-1 (FHV-1) and in Marek's Disease Virus (MDV) of chickens. We have identified the MHC class I immune evasion molecule in FHV-1 and have cloned and analysed this gene, and are constructing mutant viruses that lack the immune evasion molecule. We have also tentatively identified the immune evasion molecule of MDV and are cloning and analyzing this gene as well. (3) I am mentoring two full-time Ph.D. students (Jerome Moulden and Ram Kamal), as well as a Ph.D. rotation student (Likit Preeyanon, Fall '08) and two undergraduate students, Cari Hearn (in a CVM Summer Research Program, Summer '08) and Eric Collins (as a Professorial Assistant, Fall '08 - Spring '09). (4) Events include attending the CMB/GEN Retreat at the Kellogg Biological Station (Aug '08) and the Chicago Autumn Immunology Conference (Nov '08). (5) Services include teaching classes for MMG813 (Graduate Virology), MMG101 (Preview of Microbiology), MMG103 (Frontiers of Microbiology and Molecular Genetics), and MMG559 (Veterinary Microbiology & Immunology) PARTICIPANTS: Individuals: Ian York: Principle investigator Barbara Christian: Research technician Training/Professional development: Jerome Moulden: Graduate student Ram Kamal: Graduate student. Collaborators: Roger Maes (DCPAH, MSU) Henry Hunt (ADOL, MSU) TARGET AUDIENCES: EFFORTS include mentoring two full-time Ph.D. students (Jerome Moulden and Ram Kamal), as well as a Ph.D. rotation student (Likit Preeyanon, Fall '08) and two undergraduate students, Cari Hearn (in a CVM Summer Research Program, Summer '08) and Eric Collins (as a Professorial Assistant, Fall '08 - Spring '09). I have also taught classes for MMG813 (Graduate Virology, Spring '08), MMG101 (Preview of Microbiology (Fall '07, Fall '08)), MMG103 (Frontiers of Microbiology and Molecular Genetics (Spring '07, Spring '08)), and MMG559 (Veterinary Microbiology & Immunology (Fall '08)) PROJECT MODIFICATIONS: (1) We have focused on the autoimmune disease akylosing spondylitis as a model for aminopeptidase effects on immunodominance; this change was based on new published research indicating an important role for these aminopeptidases in pathogenesis of disease and in susceptibility to disease. As part of this new focus we have set aside research on alternate ER-localized aminopeptidases and focused on the previously described aminopeptidase ERAP1. (2) In viral pathogenesis we have added the virus Marek's Disease Virus as a well-characterized virus that inhibits MHC class I antigen presentation. This will enable us to use chickens as a model for viral pathogenesis and vaccine development. We also plan to use infectious laryngotracheitis virus (ILTV), also a pathogen of chickens, in order to use the efficient chicken model. As well as acting as models for herpesvirus infections of other domestic animals and of humans, both MDV and ILTV are important pathogens of chickens.
Impacts
IMPACTS (1) We have learned that aminopeptidase allelic variants have different peptide substrate specificities, potentially leading to differences in antigen presentation and T cell recognition, resulting in changes in immunodominance in the presence of different aminopeptides alleles. We will continue to analyze this in the context of susceptibility to autoimmune diseases, especially ankylosing spondylitis. (2) We have identified an MHC class I immune evasion gene of feline herpesvirus 1 (FHV-1) and characterized the gene by cloning and transfection in cultured cells. We are in the process of constructing a knockout virus lacking this gene in order to understand the function of this gene in viral pathogenesis and immunity to viruses. We will test the efficacy of this knockout virus as a backbone for vaccines against FHV-1, predicting that reducing immune evasion will result in a highly antigenic, attenuated virus. (3) We have identified an MHC class I immune evasion gene of Marek's Disease Virus (MDV) of chickens and are in the process of characterizing the gene by cloning and transfection in cultured cells. We will construct a knockout virus lacking this gene in order to understand the function of this gene in viral pathogenesis and immunity to viruses. We will test the efficacy of this knockout virus as a backbone for vaccines against MDV, predicting that reducing immune evasion will result in a highly antigenic, attenuated virus.
Publications
- 3. Hearn, A., York, I.A., and Rock, K.L. 2008. The Specificity of Trimming of MHC Class I-Presented Peptides in the Endoplasmic Reticulum. J. Immunol. (Submitted, in revision)
- 1. Towne, C. F., York, I. A., Watkin, L. B., Lazo, J. S., and Rock, K. L. (2007). Analysis of the role of bleomycin hydrolase in antigen presentation and the generation of CD8 T cell responses. J Immunol 178, 6923-6930.
- 2. Towne, C., F., York, I., A., Neijssen, J., Karow, M., L., Murphy, A., J., Valenzuela, D., M., Yancopoulos, G., D., Neefjes, J., J., and Rock, K., L. (2008). Puromycin-Sensitive Aminopeptidase Limits MHC Class I Presentation in Dendritic Cells but Does Not Affect CD8 T Cell Responses during Viral Infections. J Immunol 180, 1704-1712.
- 4. Kawahara, M., York, I.A., Hearn, A., and Rock, K.L. 2008. Analysis of the role of tripeptidyl peptidase II in MHC class I antigen presentation in vivo. J. Immunol. (Submitted, in revision)
- 5. Hearn, A., York, I.A., and Rock, K.L. 2008. Characterizing the Specificity and Co-operation of Aminopeptidases in the Cytosol and ER in MHC Class I antigen Presentation. (In preparation)
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