Experimental study on targeted of multiple glioma-associated antigens sensitized dentritic cell activated cytotoxic T lymphocytes targeting on malignant glioma
10.13699/j.cnki.1001-6821.2017.05.017
- VernacularTitle:恶性胶质瘤相关抗原表位肽激活的树突状细胞致敏的细胞毒性T淋巴细胞靶向治疗恶性胶质瘤的实验研究
- Author:
Yun ZHANG
1
;
Xi-Luan JI
;
Zhao-Xia LUO
;
Shun YANG
;
Liang XIE
;
Wen-Wen ZOU
;
Bing-Feng LIU
;
Shu JIANG
Author Information
1. 深圳市茵冠生物科技有限公司
- Keywords:
dentritic cell;
cytotoxic T lymphocyte;
glioma
- From:
The Chinese Journal of Clinical Pharmacology
2017;33(5):448-451,455
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the cytotoxicity of multiple gliomaassociated antigens sensitized dentritic cell activated cytotoxic T lymphocytes (GDC-CTL) on the human glioma cell line U87 in vitro and the anti-tumor effect of GDC-CTL on the BALB/c nude mouse model of malignant glioma in vivo.Methods Multiple glioma-associated antigens sensitized dentritic cell (GDC) and GDC-CTL were prepared and then analyzed with the phenotypes by flow cytometry.Cytotoxicity of GDC-CTL on U87 cells was determined by CCK8 assay and the level of interferon-γ (IFN-γ) secreted from GDC-CTL co-culturing with U87 cells for 48 h was detected by ELISA at different effect/target ratios (5∶ 1,10∶1,20∶1).The T lymphocytes without activation with GDC were evaluated as the control group.The BALB/c Nude mice tumor model established by the subcutaneous injection of U87 cells was adopted to assess the anti-tumor effect.The mice were randomly divided into four groups:the control group receiving subcutaneous injection with 0.9％ NaCl 0.2 mL,the model,intravenous treatment and local treatment groups receiving subcutaneous injection with 1 × 107 U87 cells in 0.2 mL Dulbecco's Modified Eagle Medium (DMEM).When the diameter of tumor tissue reached 3 mm,the model group was subcutaneously injected with 0.9％ NaC1 0.2 mL surrounding the tumor,while the intravenous treatment group and local treatment group were injected with 0.2 × 107 GDC-CTL in 0.2 mL phosphate buffer saline (PBS) through the tail vein and subcutaneous injection into the surrounding area of the tumor respectively,3 times a week for 2 weeks.The tumor volume was calculated and the pathological changes in the tumor tissues were observed for comparison.Results Matured GDC expressing the high levels of CD83,CD1a and HLA-DR successfully activated GDC-CTL in which 93.00％ of CD3 + T lymphocytes and 69.00％ of CD3 + CD8 + T lymphocytes were detected.In vitro experiments proved that the killing rates of GDC-CTL and T lymphocytes on U87 cells were (24.35 ±1.12)％ vs (15.21 ±0.91)％,(38.57±2.10)％ vs (23.35 ±1.30)％,(59.44±3.79)％ vs (35.23 ± 2.33) ％,and the IFN-γlevels secreted from GDC-CTL and T lymphocytes co-culturing with U87 cells were (405.36±27.65) vs (371.11 ±23.23) pg · mL-1,(1509.22 ±97.16) vs (913.54 ±48.35) pg · mL-1,(2429.57 ±183.18) vs (1814.97 ± 123.24) pg · mL-1,at the different effect/target ratios of 5∶1,10∶1 and 20∶1 respectively.There were significant differences between this two groups at the effect/target ratios of 10∶1 and 20∶1 (P ＜0.05).The results obtained from the in vivo experiments showed that the tumor volumes in the intravenous treatment group and local treatment group shrank 34.83％ and 45.37％ respectively,when comparing with the model group (100.00％,P ＜ 0.05).The pathological changes of tumor tissues showed that the tumor cells in the local treatment group and intravenous treatment group were significandy decreased.Conclusion The experimental results that GDC-CTL can significantly inhibit the growth of ghoma provide more evidences to further study the effective targeting therapy on glioma.
