Induction of immunotolerance in mouse pancreatic islet transplantation through PD-1/PD-L1 costimulatory pathway
10.3760/cma.j.issn.1007-631X.2009.01.017
- VernacularTitle:通过PD-1/PD-L1共刺激通路诱导小鼠胰岛移植免疫耐受的研究
- Author:
Tao LI
;
Jiye ZHU
;
Xisheng LENG
- Publication Type:Journal Article
- Keywords:
Islets of Langerhans transplantation;
Immune tolerance;
Programmed death ligand-1;
Programmed death-1;
Costimulatory pathway;
Adenovirus vector
- From:
Chinese Journal of General Surgery
2009;24(1):45-48
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the role of recombinant adenovirus Ad-PD-L1 on immunotolerance induction in mouse pancreatic islet transplantation. Methods Full-length mouse PD-L1 cDNA linked with an internal ribosome entry site (IRES)-GFP cassette was subcloned into pShuttle-GFP-CMV( - ) shuttle plasmid. The product was cut by certain restriction endonuclease and ligate with pAdxsi vector. The adenovirus bone plasmid was transformed into DH5α competent bacteria. The positive clone was identified by restriction endonuclease digestion and further confirmed by sequencing. After linearization, the recombined adenovirus DNA was transfected into 293 cells by liposome for package and amplification, which was purified by CsC1 density gradient centrifugation. Streptozocin was injected i.p. into C57BL/6 (H-2b) mouse to induce diabetic model recipient. Recipients were randomly divided into three groups, Group A was the control. Group B and group C were injected of Ad-EGFP and Ad-PD-L1 through tail vein respectively 1 day before islet transplantation. 300 to 400 islets of DBA/2 (H-2d) were transplanted into the renal subcapsular space of the diabetic model recipient. The level of blood sugar and the graft survival time were monitored. Results Recombinant adenovirns Ad-PD-L1 have high efficiency expression of PD-L1 in recipient mouse. The survival time of grafts of Ad-PD-L1 group (27.63 ± 3. 51 ) d was significantly longer than that of the control ( 7. 85 ± 0. 33 ) d and Ad-EGFP group ( 7. 67 ± 0. 59 ) d ( P < 0. 01 ). Mixed lymphocyte response showed a specific decrease reaction of recipient lymphocyte toward donor lymphocytes. Conclusion Recombinant adenovirus Ad-PD-L1 was successfully constructed. In mouse pancreatic islet transplantation, it can suppresses the activation of recipient T lymphocyte through PD-1/PD-L1 co-stimulatory pathway, and significantly prolong the survival time of grafts.
