Most clinically relevant blood samples are characterized by a limited number of cells available for analysis combined with a low frequency of the T cell populations of interest. Current methodologies for analysis of these samples often require extensive in vitro manipulation and/or assumptions about the antigen specificity and function of the cell populations of interest. The overall goal of this project is to develop new technology that will improve ex vivo analysis of T cell specificity and function, by taking advantage of advances in bioinformatics, proteomics, protein engineering, and array technology. There are five specific aims.
Aim 1 is to improve epitope discovery practice, by developing bioinformatics-based epitope prediction algorithms, by applying recent advances in mass spectrometry to identify naturally processed MHC-bound peptides, and by measuring MHC-peptide kinetic lifetimes rather than equilibrium binding affinities.
Aim 2 is to develop novel MHC oligomers to extend tetramer staining technology to characterization of heterologous immune responses and characterization of moderate-affinity and low-avidity T cells.
Aim 3 is to develop MHCpeptide arrays and ARC arrays, The arrays will be used for functional characterization of T cells after antigen-specific capture and/or activation.
Aim 4 is to optimize ex vivo expansion of T celts using nonspecific expansion and antigen-specific stimulation protocols, and to develop new methods for antigenspecific enrichment, expansion, immortalization of T cell populations.
Aim 5 is to develop methodology for high-throughput T cell cloning and analysis, including development of microscale culture methods and application of high-throughput screening methodology to T cell characterization. Once developed and validated, these technologies will be applied to T cell identification and analysis experiments in the associated Research Projects.

Public Health Relevance

An important component of the immune system's response to viruses and other pathogens involves T lymphocytes, a kind of white blood cell. Much is known about T lymphocyte function from animal studies and from model systems, but in many cases of human disease current methodology is not suitable for the detailed mechanistic studies needed to develop new therapeutic strategies. The goal of this project is to develop new experimental tools for characterizing T lymphocytes and their responses to viral infection.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Program--Cooperative Agreements (U19)
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Special Emphasis Panel (ZAI1-KS-I)
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University of Massachusetts Medical School Worcester
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Canetta, Sarah E; Bao, Yuanyuan; Co, Mary Dawn T et al. (2014) Serological documentation of maternal influenza exposure and bipolar disorder in adult offspring. Am J Psychiatry 171:557-63
Thompson, Mikayla R; Sharma, Shruti; Atianand, Maninjay et al. (2014) Interferon ?-inducible protein (IFI) 16 transcriptionally regulates type i interferons and other interferon-stimulated genes and controls the interferon response to both DNA and RNA viruses. J Biol Chem 289:23568-81
Mathew, Anuja; Townsley, Elizabeth; Ennis, Francis A (2014) Elucidating the role of T cells in protection against and pathogenesis of dengue virus infections. Future Microbiol 9:411-25
Yin, Liusong; Trenh, Peter; Guce, Abigail et al. (2014) Susceptibility to HLA-DM protein is determined by a dynamic conformation of major histocompatibility complex class II molecule bound with peptide. J Biol Chem 289:23449-64
Parra, Miguel; Herrera, Daniel; Calvo-Calle, J Mauricio et al. (2014) Circulating human rotavirus specific CD4 T cells identified with a class II tetramer express the intestinal homing receptors ?4?7 and CCR9. Virology 452-453:191-201
Yin, Liusong; Stern, Lawrence J (2014) A novel method to measure HLA-DM-susceptibility of peptides bound to MHC class II molecules based on peptide binding competition assay and differential IC(50) determination. J Immunol Methods 406:21-33
Schmidt, Madelyn R; McGinnes-Cullen, Lori W; Kenward, Sarah A et al. (2014) Modification of the respiratory syncytial virus f protein in virus-like particles impacts generation of B cell memory. J Virol 88:10165-76
Terajima, Masanori; Co, Mary Dawn T; Ennis, Francis A (2014) Age and different influenza viruses. Lancet Infect Dis 14:101
Outinen, T K; Mäkelä, S; Huttunen, R et al. (2014) Urine soluble urokinase-type plasminogen activator receptor levels correlate with proteinuria in Puumala hantavirus infection. J Intern Med 276:387-95
Co, Mary Dawn T; Terajima, Masanori; Thomas, Stephen J et al. (2014) Relationship of preexisting influenza hemagglutination inhibition, complement-dependent lytic, and antibody-dependent cellular cytotoxicity antibodies to the development of clinical illness in a prospective study of A(H1N1)pdm09 Influenza in children. Viral Immunol 27:375-82

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