Isolation and characterization of endothelial cell growth factor from Bartonella henselae

1.研制了亨氏巴尔通体液体培养基并对其进行了评价。结果表明：1.在布氏杆菌发酵液中添加Hernin和FeSO4生长效果最好，说明铁在鸡种布鲁氏菌的生长中起着重要作用。本研究试图从亨氏假单胞菌的培养上清中分离内皮细胞生长因子。据估计，该生长因子是一种相对分子质量约为3万的蛋白质。我们用灌注层析法得到了在盐浓度相对较高的区域洗脱的高活性组分。然而，由于操作过程中的失活，该生长因子的充分纯化尚未完成。亨氏假单胞菌可诱导内皮细胞和单核细胞产生IL-8和MCP-1。这一效应不仅被活细菌证明，也被灭活细菌证明。此外，煮沸或多粘菌素B处理不能抑制这一效应。内皮细胞生长因子可能是亨氏杆菌分泌的一种热敏蛋白，而IL-8/MCP-1刺激因子可能是该菌的耐热成分。这些因素可能在亨氏巴氏杆菌感染的发病机制中起重要作用。冬虫夏草能抑制外周血中性粒细胞的凋亡。亨氏假单胞菌的培养液也证明了这种活性。通过灌流层析，得到了抑制中性粒细胞凋亡的单一组分。与亨氏假单胞菌的作用有关，牙龈卟啉单胞菌还能诱导内皮细胞产生IL-8/MCP-1和ICAM-1。即使是灭活的细菌也能观察到这种活性，并且不能通过添加多粘菌素B或煮沸来消除。菌毛不参与亨氏杆菌的刺激作用，包括诱导IL-8/MCP-1、内皮细胞增殖和抑制中性粒细胞的凋亡。

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.