Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada.
PLoS Pathog. 2013;9(8):e1003590. doi: 10.1371/journal.ppat.1003590. Epub 2013 Aug 29.
The adhesion of Plasmodium falciparum-infected erythrocytes (IRBC) to receptors on different host cells plays a divergent yet critical role in determining the progression and outcome of the infection. Based on our ex vivo studies with clinical parasite isolates from adult Thai patients, we have previously proposed a paradigm for IRBC cytoadherence under physiological shear stress that consists of a recruitment cascade mediated largely by P-selectin, ICAM-1 and CD36 on primary human dermal microvascular endothelium (HDMEC). In addition, we detected post-adhesion signaling events involving Src family kinases and the adaptor protein p130CAS in endothelial cells that lead to CD36 clustering and cytoskeletal rearrangement which enhance the magnitude of the adhesive strength, allowing adherent IRBC to withstand shear stress of up to 20 dynes/cm². In this study, we addressed whether CD36 supports IRBC adhesion as part of an assembly of membrane receptors. Using a combination of flow chamber assay, atomic force and confocal microscopy, we showed for the first time by loss- and gain-of function assays that in the resting state, the integrin α₅β₁ does not support adhesive interactions between IRBC and HDMEC. Upon IRBC adhesion to CD36, the integrin is recruited either passively as part of a molecular complex with CD36, or actively to the site of IRBC attachment through phosphorylation of Src family kinases, a process that is Ca²⁺-dependent. Clustering of β1 integrin is associated with an increase in IRBC recruitment as well as in adhesive strength after attachment (∼40% in both cases). The adhesion of IRBC to a multimolecular complex on the surface of endothelial cells could be of critical importance in enabling adherent IRBC to withstand the high shear stress in the microcirculations. Targeting integrins may provide a novel approach to decrease IRBC cytoadherence to microvascular endothelium.
疟原虫感染的红细胞(IRBC)与不同宿主细胞上的受体的黏附在决定感染的进展和结果方面起着不同但关键的作用。基于我们对来自成年泰国患者的临床寄生虫分离株的离体研究,我们之前提出了一个在生理切变应力下 IRBC 细胞黏附的范例,该范例由 P 选择素、ICAM-1 和 CD36 介导的募集级联组成,主要存在于原代人真皮微血管内皮细胞(HDMEC)上。此外,我们检测到内皮细胞中涉及Src 家族激酶和衔接蛋白 p130CAS 的黏附后信号事件,这些事件导致 CD36 聚集和细胞骨架重排,从而增强黏附强度,使黏附的 IRBC 能够承受高达 20 达因/平方厘米的切变应力。在这项研究中,我们探讨了 CD36 是否作为膜受体组装的一部分支持 IRBC 黏附。我们使用流动室测定、原子力和共聚焦显微镜的组合,首次通过功能丧失和获得功能测定表明,在静止状态下,整合素 α₅β₁不支持 IRBC 和 HDMEC 之间的黏附相互作用。在 IRBC 黏附到 CD36 后,整合素通过 Src 家族激酶的磷酸化被招募,要么作为与 CD36 结合的分子复合物的一部分被动招募,要么主动招募到 IRBC 附着的部位,这个过程是 Ca²⁺依赖性的。β1 整合素的聚类与 IRBC 募集的增加以及附着后的黏附强度增加(两种情况均增加约 40%)有关。IRBC 黏附到内皮细胞表面上的多分子复合物对于使黏附的 IRBC 能够承受微循环中的高切变应力可能至关重要。靶向整合素可能为减少 IRBC 对微血管内皮细胞的细胞黏附提供一种新方法。