Measurement of CD8+ and CD4+ T Cell Frequencies Specific for EBV LMP1 and LMP2a Using mRNA-Transfected DCs

• Published in: PloS one Vol. 10; no. 5; p. e0127899
• Main Authors: Sohn, Dae-Hee, Sohn, Hyun-Jung, Lee, Hyun-Joo, Lee, Seon-Duk, Kim, Sueon, Hyun, Seung-Joo, Cho, Hyun-Il, Cho, Seok-Goo, Lee, Suk-Kyeong, Kim, Tai-Gyu
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 29.05.2015; Public Library of Science (PLoS)
• Subjects: Antigen presentation; Cancer; CD4 antigen; CD4-Positive T-Lymphocytes - immunology; CD8 antigen; CD8-Positive T-Lymphocytes - immunology; Cells, Cultured; Cytokines; Cytokines - immunology; Cytokines - metabolism; Cytotoxicity; Dendritic cells; Dendritic Cells - immunology; Dendritic Cells - virology; Enzyme-Linked Immunosorbent Assay; Epitopes; Epstein-Barr Virus Infections - immunology; Helper cells; Herpesvirus 4, Human - genetics; Herpesvirus 4, Human - pathogenicity; Humans; Immune response; Immune response (cell-mediated); Immune system; Immunity, Cellular; Immunology; Immunoproliferative diseases; Infections; Interferon; Interferon-gamma - metabolism; Interleukin 2; Interleukin 4; Latent membrane protein 1; Leukocytes (mononuclear); Leukocytes, Mononuclear - immunology; LMP2A protein; Lymphoblastoid cell lines; Lymphocytes; Lymphocytes B; Lymphocytes T; Lymphoma; Lymphomas; Medical research; Medicine; Membrane proteins; mRNA; Pathogenesis; Peptides; Peripheral blood mononuclear cells; RNA, Messenger; Stem cells; Stimulators; Studies; T cell receptors; Transfection; Tumor necrosis factor-α; Viral Matrix Proteins - genetics; Viral Matrix Proteins - immunology; Virology; γ-Interferon
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0127899

An EBV-specific cellular immune response is associated with the control of EBV-associated malignancies and lymphoproliferative diseases, some of which have been successfully treated by adoptive T cell therapy. Therefore, many methods have been used to measure EBV-specific cellular immune responses. Previous studies have mainly used autologous EBV-transformed B-lymphoblastoid cell lines (B-LCLs), recombinant viral vectors transfected or peptide pulsed dendritic cells (DCs) as stimulators of CD8(+) and CD4(+) T lymphocytes. In the present study, we used an interferon-γ (IFN-γ) enzyme-linked immunospot (ELISPOT) assay by using isolated CD8(+) and CD4(+) T cells stimulated with mRNA-transfected DCs. The frequency of latent membrane protein 1 (LMP1)-specific IFN-γ producing CD4(+) T cells was significantly higher than that of LMP2a. The frequency of IFN-γ producing CD4(+) T cells was significantly correlated with that of CD8(+) T cells in LMP1-specific immune responses (r = 0.7187, Pc < 0.0001). To determine whether there were changes in LMP1- or LMP2a-specific immune responses, subsequent peripheral blood mononuclear cells (PBMCs) samples were analyzed. Significant changes were observed in 5 of the 10 donors examined, and CD4(+) T cell responses showed more significant changes than CD8(+) T cell responses. CD8(+) and CD4(+) T cells from EBV-seropositive donors secreted only the Th1 cytokines IFN-γ, TNF-α, and IL-2, while Th2 (IL-4) and Th17 (IL-17a) cytokines were not detected. CD4(+) T cells secreted significantly higher cytokine levels than did CD8(+) T cells. Analysis of EBV-specific T cell responses using autologous DCs transfected with mRNA might provide a comprehensive tool for monitoring EBV infection and new insights into the pathogenesis of EBV-associated diseases.