Zhou Hao, Ran Yali, Da Qi, Shaw Tanner S, Shang Dan, Reddy Anilkumar K, López José A, Ballantyne Christie M, Ware Jerry, Wu Huaizhu, Peng Yuandong
Department of Hospital Infection Management of Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Cardiovascular Research Section, Department of Medicine, Baylor College of Medicine, Houston, TX 77030;
Cardiovascular Research Section, Department of Medicine, Baylor College of Medicine, Houston, TX 77030;
J Immunol. 2016 Jul 1;197(1):288-95. doi: 10.4049/jimmunol.1501946. Epub 2016 May 20.
Localization of the platelet glycoprotein Ib-IX complex to the membrane lipid domain is essential for platelet adhesion to von Willebrand factor and subsequent platelet activation in vitro. Yet, the in vivo importance of this localization has never been addressed. We recently found that the disulfide linkage between Ibα and Ibβ is critical for the association of Ibα with the glycosphingolipid-enriched membrane domain; in this study, we established a transgenic mouse model expressing this mutant human Ibα that is also devoid of endogenous Ibα (HαSSMα(-/-)). Characterization of this model demonstrated a similar dissociation of Ibα from murine platelet glycosphingolipid-enriched membrane to that expressed in Chinese hamster ovary cells, which correlates well with the impaired adhesion of the transgenic platelets to von Willebrand factor ex vivo and in vivo. Furthermore, we bred our transgenic mice into an atherosclerosis-prone background (HαSSMα(-/-)ApoE(-/-) and HαWTMα(-/-)ApoE(-/-)). We observed that atheroma formation was significantly inhibited in mutant mice where fewer platelet-bound CD11c(+) leukocytes were circulating (CD45(+)/CD11c(+)/CD41(+)) and residing in atherosclerotic lesions (CD45(+)/CD11c(+)), suggesting that platelet-mediated adhesion and infiltration of CD11c(+) leukocytes may be one of the mechanisms. To our knowledge, these observations provide the first in vivo evidence showing that the membrane GEM is physiologically and pathophysiologically critical in the function of the glycoprotein Ib-IX complex.
血小板糖蛋白Ib-IX复合物定位于膜脂域对于血小板在体外黏附于血管性血友病因子及随后的血小板激活至关重要。然而,这种定位在体内的重要性从未得到探讨。我们最近发现,Ibα与Ibβ之间的二硫键对于Ibα与富含糖鞘脂的膜域的结合至关重要;在本研究中,我们建立了一种转基因小鼠模型,该模型表达这种突变的人Ibα且缺乏内源性Ibα(HαSSMα(-/-))。对该模型的表征表明,Ibα与小鼠富含糖鞘脂的血小板膜的解离与在中国仓鼠卵巢细胞中表达的情况相似,这与转基因血小板在体内外对血管性血友病因子的黏附受损密切相关。此外,我们将我们的转基因小鼠培育成易患动脉粥样硬化的背景(HαSSMα(-/-)ApoE(-/-)和HαWTMα(-/-)ApoE(-/-))。我们观察到,在突变小鼠中动脉粥样硬化形成受到显著抑制,循环中(CD45(+)/CD11c(+)/CD41(+))和驻留在动脉粥样硬化病变中(CD45(+)/CD11c(+))的血小板结合的CD11c(+)白细胞较少,这表明血小板介导的CD11c(+)白细胞的黏附和浸润可能是其中一种机制。据我们所知,这些观察结果提供了首个体内证据,表明膜糖脂筏在糖蛋白Ib-IX复合物的功能中在生理和病理生理方面都至关重要。