Isolation and characterization of endothelial cell growth factor from Bartonella henselae

汉赛巴尔通体内皮细胞生长因子的分离和表征

基本信息

批准号：
12670259
负责人：
ODA Hiroshi
金额：
$ 2.24万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670259/
关键词：
emerging infectious disease endothelial cells growth factor bacillary angiomatosis cat scratch disease Bartonella henselae zoonosis apoptosis 内皮細胞増殖因子細菌生血管腫細菌性血管種 Bartnella henselae Porphyromonas gingivalis 血管内皮細胞分離精製

项目摘要

1. A liquid growth-medium for Bartonella henselae was developed and evaluated. The best growth was observed in brucella broth with hernin and FeSO_4 suggesting that iron has the important role in the growth of B. henselae.2. We tried to isolate the endothelial cell growth factor in the culture medium of B. henselae. The growth factor was estimated to be a protein with molecular weight of about 30,000. We obtained the high activity fraction eluted in the area of comparably high salt concentration by using a perfusion chromatography. However, the sufficient purification of this growth factor was not yet completed because of inactivation during the manipulation.3. B. henselae induced the production of IL-8 and MCP-1 in endothelial cells and monocytes. This effect was demonstrated not only by live bacteria but also by inactivated bacteria. Furthermore, the treatment of boiling or Polymyxin B did not inhibit this effect.4. The endothelial cell growth factor may be a heat-sensitive protein secreted from B. henselae, while the stimulatory factor of IL-8/MCP-1 may be a heat-resistant component of this organism. These factors may play the important role in the pathogenesis of B. henselae infection.5. B. henselae inhibited the apoptosis of peripheral neutrophils. This activity was also demonstrated by the culture medium of B. henselae. We obtained the single fraction that inhibited the neutrophil apoptosis by perfusion chromatography.6. In relation to the effect of B. henselae, Porphyromonas gingivalis was also shown to induce the production of IL-8/MCP-1 and the expression of ICAM-1 in endothelial cells. This activity was observed even by inactivated bacteria, and was not abolished by addition of polymyxin B or boiling.7. Pili did not participate in the stimulatory effects of B. henselae including induction of IL-8/MCP-1, proliferation of endothelial cells, and inhibition of neutrophil apoptosis.

1。开发并评估了Bartonella henselae的液体生长中等。在布鲁氏菌肉汤中观察到了最好的生长，伴有hernin和feso_4，这表明铁在Henselae.2的生长中具有重要作用。2。我们试图分离henselae培养基中的内皮细胞生长因子。估计生长因子是一种分子量约为30,000的蛋白质。我们通过使用灌注色谱法获得了在盐浓度相对高的地区洗脱的高活性分数。但是，由于操纵过程中的失活，该生长因子的足够纯化尚未完成。3。 Henselae诱导内皮细胞和单核细胞中IL-8和MCP-1的产生。这种作用不仅通过活细菌证明了这种作用，而且还通过灭活细菌证明了这种作用。此外，沸腾或多粘质B的治疗没有抑制这种作用。4。内皮细胞生长因子可能是henselae分泌的热敏蛋白，而IL-8/MCP-1的刺激因子可能是该生物体的耐热成分。这些因素可能在Henselae感染的发病机理中起重要作用。5。 B. henselae抑制周围嗜中性粒细胞的凋亡。 Henselae的培养基也证明了这一活性。我们获得了通过灌注色谱法抑制嗜中性粒细胞凋亡的单个部分。6。关于henselae的作用，卟啉单胞菌还显示出诱导IL-8/MCP-1的产生和内皮细胞中ICAM-1的表达。即使被灭活细菌也可以观察到这种活性，并且不会通过添加多聚淋赛B或沸腾来消除这种活性。7。 PILI未参与henselae芽孢杆菌的刺激作用，包括诱导IL-8/MCP-1，内皮细胞的增殖和抑制嗜中性粒细胞凋亡。