A dendritic cell-based assay for measuring memory T cells specific to dengue envelope proteins in human peripheral blood

• Published in: Journal of virological methods Vol. 173; no. 2; pp. 175 - 181
• Main Authors: Sun, Peifang, Beckett, Charmagne, Danko, Janine, Burgess, Timothy, Liang, Zhaodong, Kochel, Tadeusz, Porter, Kevin
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 01.05.2011; Elsevier
• Subjects: Biological and medical sciences; Blood - immunology; CD4-Positive T-Lymphocytes - chemistry; CD4-Positive T-Lymphocytes - immunology; CD8-positive T-lymphocytes; CD8-Positive T-Lymphocytes - chemistry; CD8-Positive T-Lymphocytes - immunology; Cells, Cultured; Clinical Laboratory Techniques - methods; dendritic cells; Dendritic Cells - immunology; Dengue; Dengue E protein; Dengue Vaccines - immunology; Enzyme-Linked Immunospot Assay - methods; Fundamental and applied biological sciences. Psychology; Human dendritic cell; Human T cell; Humans; immune response; Immunologic Memory; Interferon ELISPOT; interferon-gamma; Interferon-gamma - secretion; Leukocyte Common Antigens - analysis; memory; Microbiology; Primates; proteins; T-Lymphocyte Subsets - immunology; Techniques used in virology; vaccines; Viral Envelope Proteins - immunology; Virology; viruses; Human T cell; Human dendritic cell; Dengue E protein; Dengue; Interferon ELISPOT; Human; Dendritic cell; Peripheral blood circulation; Method; Protein; Infection; Viral disease; Arbovirus disease; T-Lymphocyte; Interferon; ELISPOT assay
• ISSN: 0166-0934; 1879-0984; 1879-0984
• DOI: 10.1016/j.jviromet.2011.01.023

Dengue envelope (E) protein is a dominant immune inducer and E protein-based vaccines elicited partial to complete protection in non-human primates. To study the immunogenicity of these vaccines in humans, an enzyme linked immunospot (ELISPOT) assay for measuring interferon gamma (IFN-γ) production was developed. Cells from two subject groups, based on dengue-exposure, were selected for assay development. The unique feature of the IFN-γ ELISPOT assay is the utilization of dendritic cells pulsed with E proteins as antigen presenting cells. IFN-γ production, ranging from 53–513 spot forming units per million peripheral blood mononuclear cells (PBMCs), was observed in dengue-exposed subjects as compared to 0–45 IFN-γ spot forming units in dengue-unexposed subjects. Further, both CD4+ and CD8+ T cells, and cells bearing CD45RO memory marker, were the major sources of IFN-γ production. The assay allowed quantification of E-specific IFN-γ-secreting memory T cells in subjects 9 years after exposure to a live-attenuated virus vaccine and live-virus challenge. Results suggested that the dendritic cell-based IFN-γ assay is a useful tool for assessing immunological memory for clinical research.