| 赵锦荣 白海燕 郭晏海 刘永兰 颜真.35-37 kDa形式可溶性MHC I 释放机制研究[J].,2015,15(18):3415-3417 |
| 35-37 kDa形式可溶性MHC I 释放机制研究 |
| Release Mechanismof 35-37 kDa of Soluble MHC I |
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| DOI： |
| 中文关键词: 主要组织相容性复合体I 金属蛋白酶 外排小体 多泡小体 |
| 英文关键词: Major histocompatibility complex class I Metalloproteinase Exosome Multivesicular bodies |
| 基金项目:国家自然科学基金项目（31100547） |
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| 中文摘要: |
| 目的：探讨35-37 kDa 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 kDa 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 kDa 可溶性MHC I；用Quantity One
软件对43 和35-37 kDa 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 kDa 可溶性MHC I产生；蔗糖
同时显著促进43 和35-37 kDa 可溶性MHC I产生；43 kDa 可溶性MHC I存在于外排小体上，而35-37 kDa 可溶性MHC I在外
排小体上检测不到。结论：35-37 kDa 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 kDa 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 kDa 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 kDa 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 kDa
of soluble MHC I simultaneously, and sucrose could increase the release of 43 and 35-37 kDa of soluble MHC I simultaneously. No
35-37 kDa of soluble MHC I was found in exosomes, contrary to 43 kDa of soluble MHC I.Conclusion:Both 35-37 kDa of soluble
MHC I and exosomes were released by fusing of multivesicular bodies with plasma membrane, but while being released, 35-37 kDa of
soluble MHC I was independent of exosomes, while 43 kDa of soluble MHC I was located in exosomes and released with exosomes. |
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