Asialo GM1 is a receptor for Pseudomonas aeruginosa adherence to regenerating respiratory epithelial cells
Asialo GM1 is a receptor for Pseudomonas aeruginosa adherence to regenerating respiratory epithelial cells
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DOI：
10.1128/iai.64.5.1582-1588.1996
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发表时间：
1996-05-01
影响因子：
3.1
通讯作者：
Puchelle, E
中科院分区：
文献类型：
--
作者：
DeBentzmann, S;Roger, P;Puchelle, E
我们研究了 asialo GM1 作为上皮受体在铜绿假单胞菌对囊性纤维化 (CF) 和非 CF 患者再生呼吸道上皮细胞亲和力增加中的影响。从非 CF 受试者和 Delta F 508 跨膜电导调节蛋白 (CFTR) 突变纯合的 CF 患者的鼻息肉中获得人呼吸道上皮细胞，并根据外植体生长模型进行培养。在生长物的外围，观察到再生呼吸道上皮细胞以板状伪足分布在 I 型胶原蛋白基质上，这是损伤后呼吸道上皮伤口修复的特征。发现 Delta F 508 纯合 CF 组中铜绿假单胞菌对再生呼吸道上皮细胞的粘附显着高于非 CF 组 (P < 0.001)。使用针对 asialo GM1 的兔多克隆抗体进行的体外竞争性结合抑制测定表明，阻断 asialo GM1 会降低 Delta F 508 纯合 CF 培养物 (P < 0.001) 以及非 CF 培养物 (P < 0.001) 中铜绿假单胞菌对再生呼吸道上皮细胞的粘附。 CF 患者中阻断 asialo GM1 的效率显着高于非 CF 受试者 (P < 0.05)。通过预嵌入标记和免疫电子显微镜确定的asialo GM1的分布清楚地证明了asialo GM1通过再生呼吸道上皮细胞在顶端膜上的特异性表达，而其他细胞表型并不在顶端表达asialo GM1。这些结果证明(i)asialo GM1是铜绿假单胞菌的顶膜受体，在CF和非CF再生呼吸道上皮细胞表面表达，并且(ii)asialo GM1在再生呼吸道上皮中特异性恢复。这些结果表明，在 CF 中，上皮修复代表了暴露 asialo GM1 以促进铜绿假单胞菌粘附的主要事件。
We investigated the implication of asialo GM1 as an epithelial receptor in the increased Pseudomonas aeruginosa affinity for regenerating respiratory epithelial cells from cystic fibrosis (CF) and non-CF patients. Human respiratory epithelial cells were obtained from nasal polyps of non-CF subjects and of CF patients homozygous for the Delta F 508 transmembrane conductance regulator protein (CFTR) mutation and cultured according to the explant-outgrowth model. At the periphery of the outgrowth, regenerating respiratory epithelial cells spreading over the collagen I matrix with lamellipodia were observed, characteristic of respiratory epithelial wound repair after injury. P. aeruginosa adherence to regenerating respiratory epithelial cells was found to be significantly greater in the Delta F 508 homozygous CF group than in the non-CF group (P < 0.001). In vitro competitive binding inhibition assays performed with rabbit polyclonal antibody against asialo GM1 demonstrated that blocking asialo GM1 reduces P. aeruginosa adherence to regenerating respiratory epithelial cells in Delta F 508 homozygous CF cultures (P < 0.001) as well as in non-CF cultures (P < 0.001). Blocking of asialo GM1 was significantly more efficient in CF patients than in non-CF subjects (P < 0.05). Distribution of asialo GM1 as determined by preembedding labelling and immunoelectron microscopy clearly demonstrated the specific apical membrane expression of asialo GM1 by regenerating respiratory epithelial cells, whereas other cell phenotypes did not apically express asialo GM1. These results demonstrate that (i) asialo GM1 is an apical membrane receptor for P. aeruginosa expressed at the surface of CF and non-CF regenerating respiratory epithelial cells and (ii) asialo GM1 is specifically recovered in regenerating respiratory epithelium. These results suggest that in CF, epithelial repair represents the major event which exposes asialo GM1 for P. aeruginosa adherence.