PD-1/PD-L1 costimulatory pathway-induced mouse islet transplantation immune tolerance.

BACKGROUND

Programmed death-1/PD-1 ligand-1 (PD-1/PD-L1) costimulatory signals may play an important role in T-cell-induced immune response. The aim of this study is to investigate the role of the PD-1/PD-L1 costimulatory pathway on immunotolerance induction in mouse pancreatic islet transplantation.

METHODS

Full-length mouse PD-L1 cDNA was subcloned into pShuttle-GFP-CMV(-) shuttle plasmid. The product was cut by certain restriction endonuclease and ligated with pAdxsi vector. The adenovirus bone plasmid was transformed into DH5α-competent bacteria. After linearization, the recombined adenovirus DNA was transfected into 293 cells for package and amplification. Streptozotocin was injected intraperitoneally into C57BL/6 (H-2(b)) mouse to induce diabetic model recipient. Recipients were randomly divided into 3 groups. Group A was the control. Group B and group C were injected with Ad-EGFP and Ad-PD-L1 through the tail vein, respectively, 1 day before islet transplantation. The 300 to 400 islets of DBA/2 (H-2(d)) were transplanted into the renal subcapsular space of the diabetic model recipient. We monitored and analyzed the blood glucose levels and the survival time of grafts after transplantation.

RESULTS

Recombinant adenovirus Ad-PD-L1 had high efficiency expression of PD-L1 in recipient mouse. The blood glucose concentration of mice in the Ad-PD-L1 gene treatment group was obviously lower than that of the control and Ad-EGFP treatment groups and was stable and kept within the normal range at post-transplant 21 days. The survival time of grafts in the Ad-PD-L1 group (27.6 ± 3.5 days) was significantly longer than in the control (7.8 ± 0.33 days) and Ad-EGFP groups (7.6 ± 0.59 days), P < .01. Mixed lymphocyte response showed a specific decrease reaction of recipient lymphocyte vs donor lymphocyte. Flow cytometry detection showed that unsplit cells occupied 90% of recipient mouse lymphocytes, but unsplit cells among normal C57BL/6 mouse lymphocytes without Ad-PD-L1 gene treatment were 51 in the control group. The differences between them were significant (P < .01).

CONCLUSION

Recombinant adenovirus Ad-PD-L1 has been successfully constructed. In mouse pancreatic islet transplantation, it can suppress the activation of recipient T lymphocyte through the PD-1/PD-L1 costimulatory pathway, and significantly prolong the survival time of grafts.