title: An additional CD28 costimulatory signal enhances proliferation and cytotoxicity of murine T cell-derived CIK cell.
abstract: OBJECTIVE
  Cytokine induced killer (CIK) cells are ex-vivo expanded T cells endowed with both T and Natural Killer cell properties. The standard protocol for generation of CIK cells is to culture peripheral blood mononuclear cells (PBMC) in the presence of interferon- gamma (IFN-γ), monoclonal antibody (mAb) against CD3 and interleukin-2 (IL-2). However, this protocol lacks costimulatory signal (CD28), crucial for T cell activation. Herein, the proliferation and functional properties of murine thymocytes derived CIK cells generated with or without costimulatory activation provided by anti-CD28 mAb were examined.
  
  
  METHODS
  The proportion of CIK (Thy1.2+NK1.1+ and CD8+NK1.1+) cells in culture and the expression of cytotoxic granules (granzyme B and perforin) and proinflammatory cytokines (IFN-γ and tumor necrosis factor-alpha (TNF-α)) were determined by flow cytometry. Additionally, CIK cell cytotoxicity against YAC-1 murine lymphoma cells was measured by a propidium iodide-based assay.
  
  
  RESULTS
  The addition of anti-CD28 to standard CIK culture conditions increased the number of Thy1.2+ NK1.1+ and CD8+ NK1.1+ (the major effector population) cells by almost 40% and 32%, respectively. Furthermore, the cytotoxic potential of CIK cells cultured with the addition of anti-CD28 mAb was also enhanced, with a corresponding increase in CIK cells expressing granzyme B, perforin, IFN-γ and TNF-α.
  
  
  CONCLUSIONS
  The addition of anti-CD28 mAb generated more effective murine T cell-derived CIK cells.
authors.affiliation: Department of Pharmacology, Faculty of Medicine Siriraj Hospital Mahidol University, Bangkok, Thailand.
authors.forename: Sunita
authors.initials: S
authors.lastname: Timalsena
authors.affiliation: Department of Pharmacology, Faculty of Medicine Siriraj Hospital Mahidol University, Bangkok, Thailand.
authors.forename: Panwadee
authors.initials: P
authors.lastname: Pluangnooch
authors.affiliation: Department of Pharmacology, Faculty of Medicine Siriraj Hospital Mahidol University, Bangkok, Thailand.
authors.forename: Adisak
authors.initials: A
authors.lastname: Wongkajornsilp
authors.affiliation: Department of Pharmacology, Faculty of Medicine Siriraj Hospital Mahidol University, Bangkok, Thailand.
authors.forename: Kitipong
authors.initials: K
authors.lastname: Soontrapa
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D018106:CD28 Antigens / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D055612:Cytokine-Induced Killer Cells / Q000276:immunology*; D003602:Cytotoxicity, Immunologic; D005434:Flow Cytometry; D053804:Granzymes / Q000378:metabolism; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000378:metabolism; D018655:Lymphocyte Count; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D054353:Perforin / Q000378:metabolism; D015398:Signal Transduction; D060168:Thymocytes / Q000166:cytology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D018106:CD28 Antigens; D014409:Tumor Necrosis Factor-alpha; D054353:Perforin; D007371:Interferon-gamma; D053804:Granzymes