LEPTOSPIRA ICTEROHEMORRHAGIAE AND LEPTOSPIRE PEPTIDOLGYCANS INDUCE ENDOTHELIAL-CELL ADHESIVENESS FOR POLYMORPHONUCLEAR LEUKOCYTES
LEPTOSPIRA ICTEROHEMORRHAGIAE AND LEPTOSPIRE PEPTIDOLGYCANS INDUCE ENDOTHELIAL-CELL ADHESIVENESS FOR POLYMORPHONUCLEAR LEUKOCYTES
复制标题
DOI：
10.1128/iai.63.8.2995-2999.1995
复制
发表时间：
1995-08-01
影响因子：
3.1
通讯作者：
PATRIARCA, P
中科院分区：
文献类型：
--
作者：
DOBRINA, A;NARDON, E;PATRIARCA, P
我们已经研究了毒性钩端螺旋体菌株泰拉莫，血清型黄疸型，对人类嗜中性多形核白细胞（PMN）的粘附培养的人脐静脉内皮细胞（HEC）的影响。用完整或超声处理的钩端螺旋体对HEC进行选择性预处理，可导致HEC-PMN粘附的剂量和时间依赖性增加（未处理HEC的粘附率为13.2% ± 2.5%，而用10（8）个完整钩端螺旋体/ml预处理4 h的HEC的粘附率为46.3% ± 5.6%[6次实验的平均值±标准误; P < 0.001]）。相反，选择性钩端螺旋体预处理PMN或在粘附试验中加入钩端螺旋体并不改变HEC-PMN粘附，钩端螺旋体诱导内皮细胞凋亡时没有检测到HEC损伤，并被RNA和蛋白质合成抑制剂以及中性粒细胞CD 11/CD 18粘附复合物和内皮细胞-白细胞粘附分子1（ELAM-1）的单克隆抗体阻止。内皮细胞。用白细胞介素-1或革兰氏阴性细菌大肠杆菌的脂多糖（LPS）预处理HEC获得了类似的结果。用LPS抑制剂多粘菌素B抑制E. coli LPS对钩端螺旋体和白细胞介素1诱导的粘附无抑制作用。同样，钩端螺旋体LPS似乎没有作用的内皮细胞粘附的增加，因为预处理HEC与钩端螺旋体LPS提取物（苯酚-水方法）或与钩端螺旋体总脂质提取物未能诱导嗜中性粒细胞的促粘附表型。相反，从我们的钩端螺旋体中提取的肽聚糖积极刺激中性粒细胞的内皮促粘附活性（对未处理的HEC的粘附率为16.5% +/- 2.1%，对用1 μ g/ml肽聚糖预处理4小时的HEC的粘附率为51.2% +/- 2.9%; [4次实验的平均值+/-标准误; P < 0.001]）。这种肽聚糖诱导的活性被CD 11/CD 18粘附复合物和ELAM-1的单克隆抗体抑制，但不被多粘菌素B抑制。我们的结论是，从致病性钩端螺旋体的肽聚糖的分子，可以直接激活血管内皮细胞，以增加他们的嗜中性粒细胞。这些观察结果可能有助于更好地了解非革兰氏阴性菌调节局部和全身炎症反应的机制。
We have examined the effect of the virulent Leptospira interrogans strain Teramo, serotype icterohemorrhagiae, on the adherence of human neutrophilic polymorphonuclear leukocytes (PMN) to cultured human umbilical vein endothelial cells (HEC). Selective pretreatment of HEC with intact or sonicated leptospires caused a dose- and time-dependent increase of HEC-PMN adhesion (13.2% +/- 2.5% adherence to untreated HEC versus 46.3% +/- 5.6% adherence to HEC pretreated for 4 h with 10(8) intact leptospires per ml [mean +/- standard error of six experiments; P < 0.001]). In contrast, selective leptospire pretreatment of PMN or the addition of leptospires during the adherence assay did not alter HEC-PMN adherence, Leptospire induction of endothelial-cell adhesiveness occurred without detectable HEC damage and was prevented by RNA and protein synthesis inhibitors and by monoclonal antibodies to the CD11/CD18 adhesion complex of neutrophils and to the endothelial-leukocyte adhesion molecule 1 (ELAM-1) of endothelial cells. Similar results were obtained with pretreatment of HEC with interleukin-1 or with the lipopolysaccharide (LPS) of the gram-negative bacterium Escherichia coli. The possibility that contamination by the LPS of gram-negative bacteria could be involved in the induction of HEC adhesiveness was ruled out by the observation that the LPS inhibitor polymyxin B, which abolished the proadhesive effect of E. coli LPS, was ineffective in inhibiting leptospire- as well as interleukin-1-induced adherence. Similarly, leptospire LPSs seemed to have no role in the increase of endothelial-cell adhesiveness, since pretreatment of HEC with a leptospire LPS extract (phenol-water method) or with a leptospire total lipid extract failed to induce the proadhesive phenotype for neutrophils. Instead, peptidoglycans extracted from our leptospires actively stimulated the endothelial proadhesive activity for neutrophils (16.5% +/- 2.1% adherence to untreated HEC versus 51.2% +/- 2.9% adherence to HEC pretreated for 4 h with 1 mu g of peptidoglycan per ml; [mean +/- standard error of four experiments; P < 0.001]). This peptidoglycan-induced activity was inhibited by monoclonal antibodies to the CD11/CD18 adhesion complex and to ELAM-1 but not by polymyxin B. We conclude that peptidoglycans from pathogenic leptospires are among the molecules that can directly activate vascular endothelial cells to increase their adhesiveness for neutrophilic granulocytes. These observations may contribute to a better understanding of the mechanisms whereby non-gramnegative bacteria modulate the local and systemic inflammatory reaction.