Open AccessMeasurement of CD8+ and CD4+ T Cell Frequencies Specific for EBV LMP1 and LMP2a Using mRNA-Transfected DCs
Author(s) -
Dae-Hee Sohn,
Honglae Sohn,
Hyun-Joo Lee,
Seon-Duk Lee,
Sueon Kim,
SeungJoo Hyun,
Hyun-Il Cho,
Seok-Goo Cho,
Suk-Kyeong Lee,
TaiGyu Kim
Publication year - 2015
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0127899
Subject(s) - elispot , cd8 , immune system , t cell , transfection , cytotoxic t cell , biology , immunology , cytokine , peripheral blood mononuclear cell , microbiology and biotechnology , antigen presenting cell , cell culture , in vitro , biochemistry , genetics
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.