An additional CD28 costimulatory signal enhances proliferation and cytotoxicity of murine T cell-derived CIK cells

作者：Timalsena Sunita; Pluangnooch Panwadee; Wongkajornsilp Adisak; Soontrapa Kitipong^*

来源：Asian Pacific Journal of Allergy and Immunology, 2017, 35(2): 67-74.

DOI：10.12932/AP0774

• CD28 monoclonal antibody
• CD3 monoclonal antibody
• Cytokine-induced killer cells
• Murine thymocytes
• Immunostimulation

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-gamma), 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.
Method: The proportion of CIK (Thyl.2(+)NKI.1(+) and CD8(+)NKI.1(+)) cells in culture and the expression of cytotoxic granules (granzyme B and perforin) and proinflammatory cytokines (IFN-gamma and tumor necrosis factor-alpha (TNF-alpha)) 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 Thyl.2(+)NK1.1(+) and CD8(+)NKI.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-gamma and TNF-alpha.
Conclusions: The addition of anti-CD28 mAb generated more effective murine T cell-derived CIK cells.

• 出版日期2017-6