Alpha-galactosylceramide loaded tumor cells in combination with TLR9 agonists induce potent anti-tumor responses in a murine model of colon cancer
10.3760/cma.j.issn.0254-5101.2014.07.001
- VernacularTitle:肿瘤细胞／糖脂类分子联合 TLR 9配体治疗小鼠结肠癌的效果研究
- Author:
Tiangeng DONG
;
Tuo YI
;
Xinqiang HONG
;
Mengxuan YANG
;
Shengli LIN
;
Xingyuan XU
;
Wenxiang LI
;
Weixin NIU
- Publication Type:Journal Article
- Keywords:
iNKT cells;
Dendritic cells;
α-Galcer;
Cancer vaccine
- From:
Chinese Journal of Microbiology and Immunology
2014;(7):491-497
- CountryChina
- Language:Chinese
-
Abstract:
Objective To design a new cancer vaccine by using alpha-galactosylceramide (α-Galcer,α-GC) loaded tumor cells in combination with TLR 9 ligand and to evaluate its therapeutic effects on colon canc-er in mice.Methods MC38 cells were transfected with lentivirus (GFP-CD1d) to prepare CD1d-MC38 cells. The expression of CD1d molecules in CD1d-MC38 cells was detected by fluorescence microscopy , RT-PCR and flow cytometry.The sorted CD1d-MC38 cells were loaded with α-Galcer to prepare CD1d-MC38/α-GC complex. Flow cytometry was performed to evaluate the efficiency of combination .A mouse model of colon cancer was es-tablished to investigate the therapeutic effects of α-Galcer loaded tumor cells in combination with TLR 9 ligand ( CD1d-MC38/α-GC+CpG1826) on colon cancer in mice by analyzing tumor growth and mice survival time .Im-munohistochemical staining was used to detect CD 4+T and CD8+T infiltrating lymphocytes in tumor tissues .Re-sults The MC38 cancer cells that expressed CD 1d and GFP were successfully constructed , among which 98.10%±2.53%were positive for CD1d.Moreover, the CD1d-MC38 cells could combine with α-Galcer effec-tively in a dose and time dependent manner .Compared with PBS treated group ,α-GC treated group and TLR9 ligand treated group , the experimental vaccine strategy was sufficient to inhibit the growth of established tumors and prolong survival of tumor-bearing mice (P<0.01).Immunohistochemistry analysis revealed that levels of CD4+T cells and CD8+T cells in experiment group were significantly higher than those in groups treated with PBS,α-GC and TLR9 ligand (P<0.01).Conclusion CD1d-MC38/α-GC in combination with CpG1826 could efficiently inhibit the growth of established tumors and prolong survival of tumor-bearing mice .Immunohisto-chemistry analysis revealed that CD 4+T cells and CD8+T cells played important roles in anti-tumor immunity.
