Anti-tumor efficiency of cytotoxic T-lymphocyte induced by activated B lymphocyte after hepatocellular carcinoma alpha fetoprotein mRNA transfection
10.3760/cma.j.issn.1673-9752.2013.01.013
- VernacularTitle:肝癌细胞甲胎蛋白mRNA转染活化的B淋巴细胞诱导细胞毒性T淋巴细胞的抗肿瘤效应
- Author:
Tao HE
;
Ling ZHANG
;
Changshan HUANG
;
Hong CUI
;
Yunjian WANG
;
Feng HAN
- Publication Type:Journal Article
- Keywords:
Liver neoplasms;
Alpha fetoprotein;
Lymphocytes
- From:
Chinese Journal of Digestive Surgery
2013;(1):53-56
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the anti-tumor efficiency of cytotoxic T-lymphocyte (CTL) induced by activated B lymphocyte after hepatocellular carcinoma (HCC) alpha fetoprotein (AFP) mRNA transfection.Methods B lymphocytes were fractionated,purified and activated by recombinant human soluble sCD40L.PGEM4Z/AFP/A64-EGFP plasmid was established in vitro,mixed with polymerase T7RNA,and then transcribed into AFP mRNA with Poly (A) sequence.B lymphocytes electrotransfected with AFP mRNA were in the experimental group,B lymphocytes electrotransfected with GAPDH mRNA were in the negative control group,and untreated B lymphocytes were in the blank control group.The expressions of antigen-presenting cell (APC)markers (CD19,CD20,CD21,CD40,CD80,CD83) and major histocompatibility complex (MHC) of the 3 groups were detected.B lymphocytes of the 3 groups were cultured with T lymphocytes at ratios of 1∶40,1 ∶ 20,1∶10 and 1∶5 to induce and ampify CTL,and then the absorbance values were detected to evaluate the proliferation ability of T lymphocytes.The killing activity of CTL was investigated with HCC cell line SMMC7721 as the target cells.All data were analyzed using the paired t test,one-way analysis of variance or Tamhane's T2 test.Results The expressions of CD19,CD20,CD21,CD40,CD80 and CD83 of the experimental group were 74 ± 11,78 ±8,80 ± 10,90 ± 11,82 ± 6,56 ± 5,which were significantly higher than 51 ± 5,60 ± 7,53 ± 5,73 ± 8,50 ± 5 and 49 ± 6 of the negative control group,and 46 ± 3,54 ± 5,41 ± 3,56 ± 5,52 ± 6 and 21 ± 4 of the blank control group (t =5.302,4.812,7.627,5.932,9.142,7.813; 11.581,7.036,13.592,12.873,9.235,14.619,P ＜ 0.01).The proliferation of CTL of the experimental group was significantly higher than that in the negative control group and blank control group (t =18.203,23.714,15.062,9.417 ; 16.833,19.392,13.871,6.592,P ＜0.01).When the T lymphocytes were mixed with the HCC cell line SMMC7721 at the ratios of 40∶ 1,20∶1and 10∶1,the killing rates of HCC cells by CTL of the exprimental group were 43％ 4％,32％ ± 4％ and 22％ ±3％,which were significantly higher than 15％ ± 5％,7％ ± 3％ and 6％ ±2％ of the negative control group,and 7％±3％,8％±3％ and 9％±4％ of the blank control group (t =9.141,13.272,11.901; 14.372,12.835,9.507,P ＜ 0.01).Conclusion Activated B lymphocytes after HCC AFP mRNA transfection may effectively induce CTL to kill HCC cells.
