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参与恶性疟原虫细胞黏附的松鼠猴脑微血管内皮细胞硫酸软骨素的特性分析。

Characterisation of the chondroitin sulphate of Saimiri brain microvascular endothelial cells involved in Plasmodium falciparum cytoadhesion.

作者信息

Fusai T, Parzy D, Spillmann D, Eustacchio F, Pouvelle B, Lépolard C, Scherf A, Gysin J

机构信息

Unité de Parasitologie, IMTSSA, Boulevard Charles Livon, Jardin du Pharo, 13007, Marseille, France.

出版信息

Mol Biochem Parasitol. 2000 Apr 30;108(1):25-37. doi: 10.1016/s0166-6851(00)00199-7.

Abstract

Cytoadhesion of Plasmodium falciparum-infected erythrocytes (IRBC) to chondroitin-4-sulphate (CSA) is inhibited by soluble CSA in vitro on Saimiri brain microvascular endothelial cells (SBEC) and in vivo in P. falciparum-infected Saimiri monkeys. We tested whether the SBEC model was appropriate for studying CSA-binding IRBC using four cell lines. All SBEC expressed a chondroitin sulphate (CS), with a composition of CSA. The mean sizes of these CSA were 20.5, 22, 23, 32.5 and 36 kDa for SBEC 3A and C2, CHO, SBEC 1D and 17, respectively. We found that cytoadhesion of the Palo-Alto (FUP)1 CSA-binding phenotype, selected by panning on SBEC 17, was specifically inhibited in a dose-dependent manner by all the purified CSA. The extent of inhibition depended on the cellular origin of the tested CSA. SBEC 17 CSA was 33 times more efficient than CHO-CSA and 21 times more efficient than the 50 kDa commercial bovine trachaea CSA. Dynabeads coated with a total extract of SBEC 1D CS-proteoglycans interacted with CSA- but not with CD36- or ICAM-1-binding IRBC. These Dynabeads also interacted specifically with the PfEMP1 DBL-3 domain, on the surface of CHO transfectants, but not with the CIDR-1 domain. Thrombomodulin was involved in IRBC adhesion to all SBEC whereas CD44 was only expressed by SBEC 1D and 17. These two CSA-proteoglycans have also been detected at the surface of human endothelial cells. Thus, the two homologous models, SBEC/Saimiri sciureus, are useful and reliable tools for the evaluation of new anti-CSA adhesion treatments and anti-disease vaccines for pregnant women.

摘要

恶性疟原虫感染的红细胞(IRBC)与硫酸软骨素-4(CSA)的细胞黏附在体外对松鼠猴脑微血管内皮细胞(SBEC)以及在体内对感染恶性疟原虫的松鼠猴中均受到可溶性CSA的抑制。我们使用四种细胞系测试了SBEC模型是否适合用于研究结合CSA的IRBC。所有SBEC均表达硫酸软骨素(CS),其组成成分为CSA。对于SBEC 3A和C2、CHO、SBEC 1D和17,这些CSA的平均大小分别为20.5、22、23、32.5和36 kDa。我们发现,通过在SBEC 17上淘选而选择的帕洛阿尔托(FUP)1结合CSA表型的IRBC的细胞黏附,受到所有纯化CSA的剂量依赖性特异性抑制。抑制程度取决于测试CSA的细胞来源。SBEC 17 CSA的效率比CHO-CSA高33倍,比50 kDa商业牛气管CSA高21倍。包被有SBEC 1D CS-蛋白聚糖总提取物的磁珠与结合CSA的IRBC相互作用,但不与结合CD36或ICAM-1的IRBC相互作用。这些磁珠还与CHO转染细胞表面的PfEMP1 DBL-3结构域特异性相互作用,但不与CIDR-1结构域相互作用。血栓调节蛋白参与IRBC与所有SBEC的黏附,而CD44仅由SBEC 1D和17表达。这两种CSA-蛋白聚糖也已在人内皮细胞表面检测到。因此,这两个同源模型,即SBEC/松鼠猴,是评估针对孕妇的新型抗CSA黏附治疗和抗病疫苗的有用且可靠的工具。

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