中文摘要：

目的：探讨35-37 k Da形式的可溶性MHC I释放机制,为开展造血系统恶性肿瘤免疫干预治疗研究奠定理论基础。方法：以细胞表面标记、免疫沉淀、免疫印迹和增强化学发光法探讨EGTA和蔗糖对THP1细胞释放43和35-37 k Da可溶性MHC I的影响;以超速离心法纯化外排小体,并用免疫沉淀、免疫印迹和增强化学发光法检测43和35-37 k Da可溶性MHC I;用Quantity One软件对43和35-37 k Da可溶性MHC I进行相对定量分析。结果：EGTA同时显著抑制43和35-37 k Da可溶性MHC I产生;蔗糖同时显著促进43和35-37 k Da可溶性MHC I产生;43 k Da可溶性MHC I存在于外排小体上,而35-37 k Da可溶性MHC I在外排小体上检测不到。结论：35-37 k Da可溶性MHC I与外排小体都来源于细胞内多泡小体同质膜的溶合后释放,但35-37 k Da可溶性MHC I并不包含在外排小体的泡囊中,而是独立于外排小体释放。

英文摘要：

Objective： To investigate the release mechanism of 35-37 k Da of soluble MHC I and to lay a foundation for the immune intervention of hematopoietic malignancies. Methods： Cell surface labeling, immunoprecipitation, Western blot and enhanced chemiluminescence methods were used to investigate the effect of EGTA and sucrose on the release of 43 and 35-37 k Da of soluble MHC I in THP1 cells respectively. Exosomes were separated from the supernatant of cell culture by ultracentrifugation, and then soluble MHC I was detected with immunoprecipitation, Western blot and enhanced chemiluminescence methods. The quantity of soluble MHC I release was analyzed with Quantity One software after chemiluminescence. Results： EGTA could inhibit the release of 43 and 35-37 k Da of soluble MHC I simultaneously, and sucrose could increase the release of 43 and 35-37 k Da of soluble MHC I simultaneously. No35-37 k Da of soluble MHC I was found in exosomes, contrary to 43 k Da of soluble MHC I. Conclusions： Both 35-37 k Da of soluble MHC I and exosomes were released by fusing of multivesicular bodies with plasma membrane, but while being released, 35-37 k Da of soluble MHC I was independent of exosomes, while 43 k Da of soluble MHC I was located in exosomes and released with exosomes.