Key Laboratory for Experimental Teratology of the Ministry of Education and Biomedical Isotope Research Center, School of Basic Medical Sciences, Shandong University, Jinan 250012, China.
Int J Mol Sci. 2020 Feb 15;21(4):1315. doi: 10.3390/ijms21041315.
Non-invasively monitoring allogeneic graft rejection with a specific marker is of great importance for prognosis of patients. Recently, data revealed that IL-27Rα was up-regulated in alloreactive CD4 T cells and participated in inflammatory diseases. Here, we evaluated whether IL-27Rα could be used in monitoring allogeneic graft rejection both in vitro and in vivo. Allogeneic (C57BL/6 donor to BALB/c recipient) and syngeneic (BALB/c both as donor and recipient) skin grafted mouse models were established. The expression of IL-27Rα in grafts was detected. The radio-probe, I-anti-IL-27Rα mAb, was prepared. Dynamic whole-body phosphor-autoradiography, ex vivo biodistribution and immunofluorescence staining were performed. The results showed that the highest expression of IL-27Rα was detected in allogeneic grafts on day 10 post transplantation (top period of allorejection). I-anti-IL-27Rα mAb was successfully prepared with higher specificity and affinity. Whole-body phosphor-autoradiography showed higher radioactivity accumulation in allogeneic grafts than syngeneic grafts on day 10. The uptake of I-anti-IL-27Rα mAb in allogeneic grafts could be almost totally blocked by pre-injection with excess unlabeled anti-IL-27Rα mAb. Interestingly, we found that I-anti-IL-27Rα mAb accumulated in allogeneic grafts, along with weaker inflammation earlier on day 6. The high uptake of I-anti-IL-27Rα mAb was correlated with the higher infiltrated IL-27Rα positive cells (CD3/CD68) in allogeneic grafts. In conclusion, IL-27Rα may be a novel molecular imaging marker to predict allorejection.
用特异性标志物无创性监测同种异体移植物排斥反应对患者的预后具有重要意义。最近的数据表明,IL-27Rα在同种反应性 CD4 T 细胞中上调,并参与炎症性疾病。在这里,我们评估了 IL-27Rα是否可用于体外和体内监测同种异体移植物排斥反应。建立了同种异体(C57BL/6 供体至 BALB/c 受体)和同基因(BALB/c 均为供体和受体)皮肤移植小鼠模型。检测移植物中 IL-27Rα的表达。制备放射性探针 I-抗 IL-27Rα mAb。进行动态全身磷自动放射性照相术、离体生物分布和免疫荧光染色。结果显示,移植后第 10 天(同种异体排斥的高峰期)同种异体移植物中检测到 IL-27Rα的最高表达。成功制备了具有更高特异性和亲和力的 I-抗 IL-27Rα mAb。全身磷自动放射性照相术显示,移植后第 10 天,同种异体移植物的放射性摄取高于同基因移植物。用过量未标记的抗 IL-27Rα mAb 预先注射可几乎完全阻断 I-抗 IL-27Rα mAb 在同种异体移植物中的摄取。有趣的是,我们发现 I-抗 IL-27Rα mAb 更早于第 6 天在同种异体移植物中积累,同时炎症较弱。I-抗 IL-27Rα mAb 的高摄取与同种异体移植物中浸润的 IL-27Rα 阳性细胞(CD3/CD68)较高有关。总之,IL-27Rα 可能是预测同种异体排斥反应的新型分子成像标志物。