Research on allogeneic mouse spleen T lymphocyte apoptosis induced by FasL transfected dendritic cells
- VernacularTitle:转染FasL基因树突状细胞诱导异基因小鼠脾脏T细胞凋亡的研究
- Author:
Weihua FU
;
Na ZHAO
;
Yujie QIU
;
Liwei ZHU
- Publication Type:Journal Article
- Keywords:
Dendritic cell;
T lymphocyte;
Apoptosis
- From:
Chinese Journal of Microbiology and Immunology
2008;28(2):97-101
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish the mouse bone marrow-derived dendritic cells expressing FasL protein and explore the mechanism of inducing allogeneic mouse spleen T lymphocyte apoptosis. MethodsMouse myeloid DCs were cultured in selective medium zontaining essential cytokines for DC growth in vitro. The mouse DCs were transfected with liposome-mediated FasL gene. The levels of FasL mRNA before and after transfection were assayed by real-time quantitative PCR. The expression levels of FasL protein were assayed by flow cytometry(FCM)and Western blot. Non-transfected DC,empty plasmid transfected DC and FasL transfected DC were infused intravenouslY into allogeneic mouse. After 7 days, the apoptosis in spleen T lymphocytes was evaluated by TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling)method and FCM. ResultsCultured in vitro, the mature myeloid DCs from mouse could be obtained.The expressions of FasL mRNA and protein in FasL transfected DCs were significantly higher. Through the detection of spleen T lymphocyte apoptosis with TUNEL,the apoptosis index(AI)was higher in FasL transfected DC(11.67±1.53),compared with non-transfected DC(2.67±0.58)and empty plasmid transfected DC(3.33±0.58),P<0.01. ConclusionA large quantity of myeloid DCs can be obtained through in vitro culture in selective medium. The liposome-mediated FasL gene transfected DCs could successfully express high levels of FasL protein. Intravenous infusion of FasL gene transfected DCs could induce apoptosis of allogeneic mouse spleen T lymphocytes.