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重组百日咳博德特氏菌 pertactin、菌毛 2 和菌毛 3 的生产与特性。

Production and characterization of recombinant pertactin, fimbriae 2 and fimbriae 3 from Bordetella pertussis.

机构信息

Department of serum, National Institute for the Control of Pharmaceutical and Biological Products, Temple of Heaven, Beijing 100050, PR China.

出版信息

BMC Microbiol. 2009 Dec 29;9:274. doi: 10.1186/1471-2180-9-274.

Abstract

BACKGROUND

Bordetella pertussis is a causative agent of pertussis or whooping cough in humans. Pertactin (Prn), fimbriae 2 (Fim2) and fimbriae 3 (Fim3) of B. pertussis are important virulence factors and immunogens which have been included in some acellular pertussis vaccines. In this present study, we cloned, expressed and purified Prn, Fim2 and Fim3, respectively. The immunogenicity and protective efficacy of the three recombinant proteins (rPrn, rFim2 and rFim3) were investigated in mouse model.

RESULTS

Three recombinant proteins with amount of 12 to 25 mg/L were produced. Compared to the control mice only immunized with adjuvant, serum IgG antibody responses were significantly induced in the mice immunized with rPrn, rFim2 or rFim3 (P < 0.001 for all three proteins). Furthermore, T cell responses characteristic of increased production of IL-2 and TNF-alpha (only for rPrn) were elicited in the mice immunized with the three proteins (P < 0.05 for all three proteins). Immunization with rPrn, but not with rFim2 or rFim3, significantly enhanced clearance of bacteria in the lungs of mice after intranasal challenge with B. pertussis (P < 0.05). When tested in a lethal intracerebral infection model, certain protection was observed in mice immunized with rPrn.

CONCLUSIONS

We have developed an efficient method to produce large amounts of rPrn, rFim2, and rFim3 from B. pertussis. The three recombinant proteins induced both humoral and cellular immune responses in mice. Immunization with rPrn also conferred protection against pertussis in mouse infection models. Our results indicated that the recombinant proteins still retain their immunological properties and highlighted the potential of the recombinant proteins for the future development of the B. pertussis vaccines.

摘要

背景

百日咳博德特氏菌是人类百日咳或百日咳的病原体。百日咳博德特氏菌的 pertactin(Prn)、菌毛 2(Fim2)和菌毛 3(Fim3)是重要的毒力因子和免疫原,已被纳入一些无细胞百日咳疫苗中。在本研究中,我们分别克隆、表达和纯化了 Prn、Fim2 和 Fim3。在小鼠模型中研究了这三种重组蛋白(rPrn、rFim2 和 rFim3)的免疫原性和保护效力。

结果

三种重组蛋白的产量为 12 至 25mg/L。与仅用佐剂免疫的对照小鼠相比,rPrn、rFim2 或 rFim3 免疫的小鼠血清 IgG 抗体反应明显增强(三种蛋白均 P<0.001)。此外,三种蛋白均能诱导 T 细胞产生特征性的 IL-2 和 TNF-α(仅针对 rPrn)反应(三种蛋白均 P<0.05)。与 rFim2 或 rFim3 免疫不同,rPrn 免疫显著增强了小鼠肺部细菌的清除(鼻内接种百日咳博德特氏菌后 P<0.05)。在致死性脑内感染模型中进行测试时,rPrn 免疫的小鼠观察到一定的保护作用。

结论

我们已经开发了一种从百日咳博德特氏菌中大量生产 rPrn、rFim2 和 rFim3 的有效方法。三种重组蛋白在小鼠中诱导了体液和细胞免疫反应。rPrn 免疫也为小鼠感染模型中的百日咳提供了保护。我们的结果表明,重组蛋白仍保留其免疫原性,并强调了重组蛋白在未来百日咳疫苗开发中的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fda0/2807877/e2d70b051074/1471-2180-9-274-1.jpg

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