Peptide-MHC Cellular Microarray with Innovative Data Analysis System for Simultaneously Detecting Multiple CD4 T-Cell Responses

• Published in: PloS one Vol. 5; no. 6; p. e11355
• Main Authors: Ge, Xinhui, Gebe, John A., Bollyky, Paul L., James, Eddie A., Yang, Junbao, Stern, Lawrence J., Kwok, William W.
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 28.06.2010; Public Library of Science (PLoS)
• Subjects: Acids; Amino Acid Sequence; Antibodies; Antigenic determinants; Antigens; Antiviral agents; Autoimmunity; Batch processing; Beta cells; CD11a antigen; CD28 antigen; CD4 antigen; CD4-Positive T-Lymphocytes - immunology; CD4-Positive T-Lymphocytes - metabolism; Correlation analysis; Cytokines; Data analysis; Data processing; Diabetes; Diabetes and Endocrinology/Type 1 Diabetes; Diabetes mellitus; Diabetics; Epitopes; Epitopes, T-Lymphocyte - immunology; Fabrication; Flow cytometry; Functional testing; Heterogeneity; HLA Antigens - immunology; HLA Antigens - metabolism; Human subjects; Humans; Immune response; Immunologic factors; Immunology/Immune Response; Immunology/Leukocyte Activation; Influenza A; Influenza A virus - immunology; Information management; Insulin; Interferon-gamma - biosynthesis; Lymphocytes; Lymphocytes T; Major histocompatibility complex; Molecular Sequence Data; Pandemics; Peptides; Peptides - chemistry; Peptides - metabolism; Phosphatase; Populations; Protein Array Analysis; Proteins; Statistical analysis; Statistical methods; Swine flu; T cell receptors; T cells; Viral antigens; United States > US; Virginia
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0011355

Peptide:MHC cellular microarrays have been proposed to simultaneously characterize multiple Ag-specific populations of T cells. The practice of studying immune responses to complicated pathogens with this tool demands extensive knowledge of T cell epitopes and the availability of peptide:MHC complexes for array fabrication as well as a specialized data analysis approach for result interpretation. We co-immobilized peptide:DR0401 complexes, anti-CD28, anti-CD11a and cytokine capture antibodies on the surface of chamber slides to generate a functional array that was able to detect rare Ag-specific T cell populations from previously primed in vitro T cell cultures. A novel statistical methodology was also developed to facilitate batch processing of raw array-like data into standardized endpoint scores, which linearly correlated with total Ag-specific T cell inputs. Applying these methods to analyze Influenza A viral antigen-specific T cell responses, we not only revealed the most prominent viral epitopes, but also demonstrated the heterogeneity of anti-viral cellular responses in healthy individuals. Applying these methods to examine the insulin producing beta-cell autoantigen specific T cell responses, we observed little difference between autoimmune diabetic patients and healthy individuals, suggesting a more subtle association between diabetes status and peripheral autoreactive T cells. The data analysis system is reliable for T cell specificity and functional testing. Peptide:MHC cellular microarrays can be used to obtain multi-parametric results using limited blood samples in a variety of translational settings.