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在小鼠模型中鉴定免疫显性衣原体抗原的蛋白质组学方法。

Proteomic identification of immunodominant chlamydial antigens in a mouse model.

机构信息

ImmPORT Therapeutics, Inc./Antigen Discovery Inc., 1 Technology Dr., Suite E309, Irvine, CA 92618, USA.

出版信息

J Proteomics. 2012 Dec 21;77:176-86. doi: 10.1016/j.jprot.2012.08.017. Epub 2012 Aug 31.

Abstract

Chlamydia trachomatis is the most common bacterial sexually transmitted pathogen in the world. To identify new vaccine candidates a protein microarray was constructed by expressing the open reading frames (ORFs) from Chlamydia mouse pneumonitis (MoPn). C57BL/6, C3H/HeN and BALB/c mice were immunized either intranasally or intravaginally with live MoPn elementary bodies (EB). Two additional groups were immunized by the intramuscular plus subcutaneous routes with UV-treated EB, using CpG and Montanide as adjuvants to favor a Th1 response, or Alum, to elicit a Th2 response. Serum samples collected from the three strains of mice were tested in the microarray. The array included the expression of 909 proteins from the 921 ORFs of the MoPn genome and plasmid. A total of 530 ORFs were recognized by at least one serum sample. Of these, 36 reacted with sera from the three strains of mice immunized with live EB. These antigens included proteins that were previously described as immunogenic such as MOMP and HSP60. In addition, we uncovered new immunogens, including 11 hypothetical proteins. In summary, we have identified new immunodominant chlamydial proteins that can be tested for their ability to induce protection in animal models and subsequently in humans.

摘要

沙眼衣原体是世界上最常见的细菌性性传播病原体。为了鉴定新的疫苗候选物,我们构建了一个蛋白质微阵列,通过表达来自鼠肺炎衣原体(MoPn)的开放阅读框(ORFs)。C57BL/6、C3H/HeN 和 BALB/c 小鼠分别通过鼻腔内或阴道内接种活 MoPn 原体(EB)进行免疫。另外两组小鼠通过肌肉内加皮下途径用经 UV 处理的 EB 进行免疫,使用 CpG 和 Montanide 作为佐剂来促进 Th1 反应,或 Alum 来引发 Th2 反应。从三种小鼠中收集的血清样本在微阵列中进行了测试。该阵列包括 MoPn 基因组和质粒的 921 个 ORFs 中的 909 种蛋白质的表达。共有 530 个 ORF 被至少一个血清样本识别。其中,36 个与用活 EB 免疫的三种小鼠的血清反应。这些抗原包括先前被描述为免疫原性的蛋白质,如 MOMP 和 HSP60。此外,我们还发现了新的免疫原,包括 11 种假设蛋白。总之,我们已经确定了新的免疫显性衣原体蛋白,可以测试它们在动物模型中诱导保护的能力,然后在人类中进行测试。

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