title: DNA immunization of Mycobacterium tuberculosis resuscitation-promoting factor B elicits polyfunctional CD8(+) T cell responses.
abstract: OBJECTIVE
  T cell-mediated immune responses, and particularly activation of polyfunctional T cells that simultaneously produce multiple cytokines, are necessary for the control of Mycobacterium tuberculosis. In the present study, we examined if DNA immunization of Mycobacterium tuberculosis resuscitation-promoting factor B (RpfB) elicits polyfunctional T cell responses in mice.
  
  
  METHODS
  C57BL/6 mice were immunized intramuscularly three times, at 3-week intervals, with RpfB-expressing plasmid DNA. For comparison, protein immunization was performed with recombinant RpfB in control mice. After immunization, RpfB-specific T cell responses were assessed by interferon-γ (IFN-γ) enzyme-linked immunosorbent spot assay and intracellular cytokine staining (ICS), and T cell polyfunctionality was assessed from the ICS data.
  
  
  RESULTS
  RpfB DNA immunization induced not only humoral immune responses, but also CD8(+) and CD4(+) T cell responses. Immunodominant T-cell epitopes were identified within RpfB by assays with overlapping peptides. RpfB DNA immunization elicited a polyfunctional CD8(+) T cell response that was dominated by a functional phenotype of IFN-γ(+)/TNF-α(+)/IL-2(-)/CD107a(+).
  
  
  CONCLUSIONS
  RpfB DNA immunization elicits polyfunctional CD8(+) T cell responses, suggesting that RpfB DNA immunization might induce protective immunity against tuberculosis.
authors.affiliation: Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon, Korea.
authors.forename: Jino
authors.identifier: 0000-0003-2132-4963
authors.initials: J
authors.lastname: Lee
authors.affiliation: Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon, Korea.
authors.forename: Jihye
authors.identifier: 0000-0001-6297-4562
authors.initials: J
authors.lastname: Kim
authors.affiliation: Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon, Korea.
authors.forename: Jeewon
authors.identifier: 0000-0001-7457-2464
authors.initials: J
authors.lastname: Lee
authors.affiliation: Department of Microbiology and Institute of Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: Sung Jae
authors.identifier: 0000-0003-0854-4582
authors.initials: SJ
authors.lastname: Shin
authors.affiliation: Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon, Korea.
authors.forename: Eui-Cheol
authors.identifier: 0000-0002-6308-9503
authors.initials: EC
authors.lastname: Shin
keywords: DNA vaccines; Polyfunctional T cells; Resuscitation-promoting factor B; Tuberculosis