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中国致病性钩端螺旋体ompL1基因的特性及OmpL1蛋白的交叉免疫原性

Characterization of the ompL1 gene of pathogenic Leptospira species in China and cross-immunogenicity of the OmpL1 protein.

作者信息

Dong Haiyan, Hu Ye, Xue Feng, Sun Dexter, Ojcius David M, Mao Yafei, Yan Jie

机构信息

Division of Basic Medical Microbiology, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital of Medical College, Zhejiang University, Hangzhou Zhejiang, PR China.

出版信息

BMC Microbiol. 2008 Dec 17;8:223. doi: 10.1186/1471-2180-8-223.

Abstract

BACKGROUND

The usefulness of available vaccine and serological tests for leptospirosis is limited by the low cross-reactivity of antigens from numerous serovars of pathogenic Leptospira spp. Identification of genus-specific protein antigens (GP-Ag) of Leptospira would be important for development of universal vaccines and serodiagnostic methods. OmpL1, a transmembrane porin of pathogenic leptospires, was identified as a possible GP-Ag, but its sequence diversity and immune cross-reactivity among different serovars of pathogenic leptospires remains largely unknown.

RESULTS

PCR analysis demonstrated that the ompL1 gene existed in all 15 official Chinese standard strains as well as 163 clinical strains of pathogenic leptospires isolated in China. In the standard strains, the ompL1 gene could be divided into three groups (ompL1/1, ompL1/2 and ompL1/3) according to their sequence identities. Immune electron microscopy demonstrated that all products of the different gene types of ompL1 are located on the surface of leptospires. The microscopic agglutination test revealed extensive yet distinct cross-immunoagglutination among the antisera against recombinant OmpL1 (rOmpL1) and leptospiral strains belonging to different ompL1 gene types. These cross-immunoreactions were further verified by ELISAs using the OmpL1 proteins as the coated antigens in serum samples from 385 leptospirosis patients. All the antisera against rOmpL1 proteins could inhibit L. interrogans strain Lai from adhering to J774A.1 cells. Furthermore, immunization of guinea pigs with each of the rOmpL1 proteins could cause cross-immunoprotection against lethal challenge with leptospires from different ompL1 gene types.

CONCLUSION

Three types of the ompL1 gene are present in pathogenic leptospires in China. OmpL1 is an immunoprotective GP-Ag which should be considered in the design of new universal vaccines and serodiagnostic methods against leptospirosis.

摘要

背景

现有钩端螺旋体病疫苗和血清学检测方法的效用受到致病性钩端螺旋体众多血清型抗原交叉反应性低的限制。鉴定钩端螺旋体属特异性蛋白抗原(GP-Ag)对于开发通用疫苗和血清诊断方法至关重要。外膜脂蛋白L1(OmpL1)是致病性钩端螺旋体的一种跨膜孔蛋白,被确定为一种可能的GP-Ag,但其在致病性钩端螺旋体不同血清型之间的序列多样性和免疫交叉反应性仍 largely未知。

结果

PCR分析表明,ompL1基因存在于所有15株中国官方标准菌株以及163株在中国分离的致病性钩端螺旋体临床菌株中。在标准菌株中,ompL1基因根据其序列同一性可分为三组(ompL1/1、ompL1/2和ompL1/3)。免疫电镜显示,ompL1不同基因类型的所有产物都位于钩端螺旋体表面。显微镜凝集试验显示,针对重组OmpL1(rOmpL1)的抗血清与属于不同ompL1基因类型的钩端螺旋体菌株之间存在广泛但明显的交叉免疫凝集。这些交叉免疫反应通过ELISA进一步验证,ELISA使用OmpL1蛋白作为包被抗原,检测385例钩端螺旋体病患者血清样本。所有针对rOmpL1蛋白的抗血清都能抑制问号钩端螺旋体赖株黏附到J774A.1细胞。此外,用每种rOmpL1蛋白免疫豚鼠可对来自不同ompL1基因类型的钩端螺旋体致死性攻击产生交叉免疫保护。

结论

中国致病性钩端螺旋体中存在三种类型的ompL1基因。OmpL1是一种免疫保护性GP-Ag,在设计针对钩端螺旋体病的新型通用疫苗和血清诊断方法时应予以考虑。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8764/2632671/328a4ed24db8/1471-2180-8-223-1.jpg

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