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Archive > Volume 22 Issue 6 > 2015,22(6):710-715. DOI:10.3872/j.issn.1007-385X.2015.06.006 Prev Next
Basic Research
Cytotoxic effect of CTLs toward melanoma A-375 cells induced by cell penetrating peptide-fused NY-ESO-1155-163
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Abstract:
Objective:To investigate the cytotoxic effect of cytotoxic lymphocytes (CTLs) activated by Tat49-57-NY-ESO-1155-163 sensitized dendritic cells on melanoma A-375 cells. Methods:We collected the peripheral blood (about 50 ml) from healthy volunteers and isolated mononuclear cells by using lymphocyte separation medium. The cells were treated with cytokines to produce dendritic cells and T lymphocytes. After sensitized with the Tat49-57-NY-ESO-1155-163 peptide, the dendritic cells were co-cultured with the T lymphocyte cells to generate antigen-specific CTLs. Phenotypes of the dendritic cells were examined by flow cytometry. The antigen-specific cytotoxic activity of the CTLs against A-375 melanoma cells in vitro was assessed by the lactate dehydrogenase (LDH) method. Human lung cancer A549 cells and leukemia K562 cells were used as controls. Results: The expression rate of CD80/CD86 in dendritic cells sensitized with Tat49-57-NY-ESO-1155-163 was (54.9±3.3)% significantly higher than these sensitized with NY-ESO-1155-163 (\[43.8±5.7\]%，P<0.05). There were also significant increase of CD40 expression (\[42.1±1.9\]% vs \[23.7±2.8\]%，P<0.05) in Tat49-57-NY-ESO-1155-163 sensitized dendritic cells. These results indicated that the Tat fragment (49-57) significantly improved the cell penetrating ability of NY-ESO-1155-163. The T lymphocytes activated by the Tat49-57-NY-ESO-1155-163 sensitized DC were mainly CD3+CD8+cells. The cytotoxic activity of the CTLs induced by Tat49-57-NY-ESO-1155-163-sensitized dendritic cells toward A-375 melanoma cells was significantly higher than these induced with NY-ESO-1155-163-sensitized dendritic cells (P<0.05). The CTLs were also specific as they killed NY-ESO-1-expressing A375 more efficient than A549 cells and K562 cells (P<0.05). Conclusion: Tat49-57 cell penetrating peptide can enhance the immunogenicity of the peptide NY-ESO-1155-163. Tat49-57-NY-ESO-1155-163 polypeptide-sensitized dendritic cells can effectively induce the specific immune response against melanoma A-375 cells.
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Project Supported:
Project supported by the Biological Treatment Demonstration Platform Project of Shanxi Province (No. 2014091105-0101)
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