Department of Immunology, Capital Medical University, Beijing, 100069, PR China.
Laboratory Department, Capital Institute of Pediatrics, Beijing 100020, PR China.
J Med Microbiol. 2012 Sep;61(Pt 9):1262-1269. doi: 10.1099/jmm.0.040907-0. Epub 2012 Jun 8.
Infections caused by Gram-negative bacteria (GNB) are increasingly common and can result in significant mortality rates due to the development of sepsis. To examine the potential usage of a recombinant Ad5-BPI(23)-Fcγ1 virus as a biological treatment against systemic infection by GNB, a construct containing the human bactericidal/permeability increasing protein (BPI) gene, encoding the functional N terminus (amino acid residues 1-199) of human BPI, and the Fcγ1 gene, encoding the Fc segment of human immunoglobulin G1, was inserted into an adenovirus serotype 5 (Ad5) chromosome to produce a recombinant Ad5-BPI(23)-Fcγ1 virus. Human A549 cells in culture and BALB/c mice were infected with the recombinant Ad5-BPI(23)-Fcγ1 virus and BPI(23)-Fcγ1 expression was confirmed by Western blot analysis and ELISA. The concentrations of BPI(23)-Fcγ1 protein were 5.59 µg ml(-1) in vitro and 21.37 ng ml(-1) in vivo and it was observed that these concentrations were sufficient to decrease endotoxin concentrations while enhancing bactericidal activity. In addition, mice treated with the recombinant Ad5-BPI(23)-Fcγ1 virus had decreased levels of IL-1β and TNF-α and were protected from an E. coli O111 : B4 challenge. Our data support the concept that Ad5-mediated BPI(23)-Fcγ1 gene delivery could be used as an ancillary biological treatment in the management of infection caused by GNB.
革兰氏阴性菌（GNB）引起的感染越来越常见，由于败血症的发展，可能导致高死亡率。为了研究一种重组 Ad5-BPI(23)-Fcγ1 病毒作为治疗革兰氏阴性菌全身感染的生物疗法的潜力，我们构建了一个包含人杀菌/通透性增加蛋白（BPI）基因的载体，该基因编码人 BPI 的功能 N 端（氨基酸残基 1-199）和 Fcγ1 基因，编码人免疫球蛋白 G1 的 Fc 片段，插入到腺病毒血清型 5（Ad5）染色体中，产生重组 Ad5-BPI(23)-Fcγ1 病毒。在体外和体内通过感染重组 Ad5-BPI(23)-Fcγ1 病毒感染 A549 细胞和 BALB/c 小鼠，并用 Western blot 分析和 ELISA 确认 BPI(23)-Fcγ1 的表达。体外 BPI(23)-Fcγ1 蛋白浓度为 5.59 µg ml(-1)，体内浓度为 21.37 ng ml(-1)，观察到这些浓度足以降低内毒素浓度，同时增强杀菌活性。此外，用重组 Ad5-BPI(23)-Fcγ1 病毒治疗的小鼠体内白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）水平降低，并能抵抗大肠杆菌 O111∶B4 的攻击。我们的数据支持这样的概念，即 Ad5 介导的 BPI(23)-Fcγ1 基因传递可以作为治疗革兰氏阴性菌感染的辅助生物疗法。
