title: Measurement of CD8+ and CD4+ T Cell Frequencies Specific for EBV LMP1 and LMP2a Using mRNA-Transfected DCs.
abstract: 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.
authors.affiliation: Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Dae-Hee
authors.initials: DH
authors.lastname: Sohn
authors.affiliation: Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Hyun-Jung
authors.initials: HJ
authors.lastname: Sohn
authors.affiliation: Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Hyun-Joo
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Seon-Duk
authors.initials: SD
authors.lastname: Lee
authors.affiliation: Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Sueon
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Seung-Joo
authors.initials: SJ
authors.lastname: Hyun
authors.affiliation: Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea, Seoul, Korea; Cancer Research Institute, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Hyun-Il
authors.initials: HI
authors.lastname: Cho
authors.affiliation: Department of Hematology, Department of Internal medicine, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Seok-Goo
authors.initials: SG
authors.lastname: Cho
authors.affiliation: Research Institute of Immunobiology, Department of Medical Lifescience, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Suk-Kyeong
authors.initials: SK
authors.lastname: Lee
authors.affiliation: Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul, Korea; Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea, Seoul, Korea; Cancer Research Institute, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Tai-Gyu
authors.initials: TG
authors.lastname: Kim
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D020031:Epstein-Barr Virus Infections / Q000276:immunology*; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000472:pathogenicity*; D006801:Humans; D007111:Immunity, Cellular; D007371:Interferon-gamma / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000276:immunology; D012333:RNA, Messenger; D014162:Transfection; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D016207:Cytokines; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D000939:Epitopes; D012333:RNA, Messenger; D014763:Viral Matrix Proteins; D007371:Interferon-gamma