Navazo M D, Daviet L, Savill J, Ren Y, Leung L L, McGregor J L
INSERM Unit 331, Faculté de Médecine RTH Laènnec, 69372 Lyon, France.
J Biol Chem. 1996 Jun 28;271(26):15381-5. doi: 10.1074/jbc.271.26.15381.
Clearance of apoptotic neutrophils by macrophages is a crucial event following the resolution of acute inflammation. CD36, together with alphavbeta3, has been identified as one of the adhesion molecules on the surface of macrophages implicated in the clearance of polymorphonuclear leukocytes. The domain on CD36 implicated in the phagocytosis of aged neutrophils remains to be elucidated. In this study, COS cells transfected with human CD36 cDNA had a significantly higher capacity to phagocytose human apoptotic neutrophils compared with murine CD36 cDNA. Moreover, monoclonal antibodies 10/5 or OKM5 (epitopes identified on amino acids 155-183) but not monoclonal antibody 13/10 (epitope identified on amino acids 30-76) inhibited phagocytosis of apoptotic neutrophils by COS cells transfected by human CD36. Swapping the human CD36 155-183 domain from human to murine CD36 (human-murine CD36 chimera) imparted to murine CD36-transfected COS cells an increased capacity to phagocytose apoptotic neutrophils. Conversely, when the murine domain 155-183 was inserted in human CD36, a decreased phagocytic capacity was observed. In addition, a synthetic peptide(155-169) but not its scrambled form significantly inhibited phagocytosis. These results identify for the first time a functional domain encompassing amino acids 155-183 on human CD36 implicated in the recognition and phagocytosis of apoptotic neutrophils.
巨噬细胞清除凋亡的中性粒细胞是急性炎症消退后的一个关键事件。CD36与αvβ3一起,已被确定为巨噬细胞表面参与清除多形核白细胞的粘附分子之一。CD36上与衰老中性粒细胞吞噬作用相关的结构域仍有待阐明。在本研究中,与转染鼠CD36 cDNA的COS细胞相比,转染人CD36 cDNA的COS细胞吞噬人凋亡中性粒细胞的能力显著更高。此外,单克隆抗体10/5或OKM5(表位在氨基酸155 - 183上)而非单克隆抗体13/10(表位在氨基酸30 - 76上)抑制了转染人CD36的COS细胞对凋亡中性粒细胞的吞噬作用。将人CD36的155 - 183结构域从人CD36交换到鼠CD36(人 - 鼠CD36嵌合体)赋予转染鼠CD36的COS细胞吞噬凋亡中性粒细胞的能力增强。相反,当将鼠的155 - 183结构域插入人CD36时,观察到吞噬能力下降。此外,一种合成肽(155 - 169)而非其乱序形式显著抑制了吞噬作用。这些结果首次确定了人CD36上一个包含氨基酸155 - 183的功能结构域,该结构域与凋亡中性粒细胞的识别和吞噬作用有关。