间充质基质细胞中NLRP3炎症小体调控红细胞血型抗体产生的机制研究

Red blood cells transfusion is a common and basic therapy in clinic. But the transfusion can unexpectedly bring about abnormal antibody against blood type antigen in many patients, which leads to annoyance to the safety and efficacy, and consequently, a waste of resources. Inflammation has been reported were involved with blood type antibody produce in recipients, however, more details in cellular and molecular mechanism on that scenario are far from totally understood. Mesenchymal stromal cells(MSCs) have been proved a vital player in inflammation response, while NOD-like receptor PYRIN domain protein 3 (NLRP3) inflammsome is a key signal trigger in the mechanism. We confirmed that NLRP3 inflammsome could be activated within bone-associated MSCs in our previously investigation. Recently we observed that NLRP3 inflammsome activated MSCs could alter the reaction between blood type antigen and antibody, thus hypothetically we believe that NLRP3 inflammsome from MSCs can impose effects on the blood type antibody produce. To validate this hypothesis, we will employ gene mutated mice to establish inflammation models to probe how NLRP3 inflammsome is involved with blood type antibody produce. Red blood cells transfusion, tack and survival, morphological analysis and chemical synthesis will be administrated in the current study. Hopefully, the present project will throw more light on how MSCs participate into blood type antibody produce through NLRP3 inflammasome. Consequently a novel small molecular target on NLRP3 inflammasome will also be synthesized to inhibit abnormal blood type antibody produce. Hopefully, after the current investigation finished, we will have novel strategy to handle with abnormal blood type antibody results from red blood cells transfusion.

红细胞输注是临床常用的基础性治疗手段，但输注后可能产生异常血型抗体，既给患者安全和疗效带来严重不利影响，也浪费血液资源。受血者机体炎症状态可调控血型抗体产生，而细胞和分子机理并未完全阐明。间充质基质细胞（Mesenchymal stromal cells, MSCs）可调控炎症免疫，NLRP3炎症小体是炎症反应的关键信号之一。我们已证实骨相关间充质基质细胞（BA-MSCs）中存在NLRP3炎症小体，发现BA-MSCs中NLRP3炎症小体活化后，可改变红细胞血型抗原反应性。故本项目拟在此基础上，以红细胞血型为研究对象，综合运用动物炎症模型的红细胞输注与功能分析、形态学以及化学合成等技术，结合体外技术手段和多种基因修饰小鼠，解析MSCs内NLRP3炎症小体在血型抗体产生中的作用，探索抑制NLRP3炎症小体来控制异常红细胞血型抗体产生的新策略，为防治红细胞输注后异常血型抗体产生奠定新的理论基础.

红细胞输注是临床常用的基础性治疗手段，但输注后可能产生异常血型抗体，既给患者安全和疗效带来严重不利影响，也浪费血液资源。受血者机体炎症状态可调控血型抗体产生，而细胞和分子机理并未完全阐明。间充质基质细胞（Mesenchymal stromal cells, MSCs）可调控炎症免疫应答。NLRP3炎症小体是炎症反应的关键信号之一，既往研究多集中在巨噬细胞、中性粒细胞等免疫细胞中。通过实施本项目研究，体外确认了MSCs中NLRP3炎症小体信号可被LPS活化；在细菌感染、炎性肠病两种小鼠模型的体内，确认了MSCs中NLRP3炎症小体信号可被活化；发现MSCs可在体内外改善体外红细胞的理化参数；观察到MSCs中NLRP3炎症小体活化后，血型抗原/抗体的反应性发生改变。合成了小分子化合物66PR，鉴定和确认了其结构和组成；证实66PR能再体外抑制MSCs细胞中NLRP3炎症小体的活化；观察到了66PR对血型抗原抗体所致红细胞破坏的抑制效应。这些研究结果，为探索抑制NLRP3炎症小体来控制异常红细胞血型抗体产生奠定了新的研究基础。