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基于人乳头瘤病毒主要外壳多表位蛋白 L1 的疫苗诱导针对沙眼衣原体生殖道感染的保护性免疫。

Protective immunity against Chlamydia trachomatis genital infection induced by a vaccine based on the major outer membrane multi-epitope human papillomavirus major capsid protein L1.

机构信息

Department of Microbiology and Immunology, Wenzhou Medical College, Wenzhou 325035, Zhejiang, China.

出版信息

Vaccine. 2011 Mar 24;29(15):2672-8. doi: 10.1016/j.vaccine.2010.12.132. Epub 2011 Feb 12.

Abstract

The administration of an efficacious vaccine is the most effective long-term measure to control the genital tract infection caused by Chlamydia trachomatis (Ct) in humans. The current challenge for Ct vaccine development is to develop an effective delivery vehicle for induction of a high level of mucosal T and complementary B cell responses. We evaluated the immunogenicity and efficacy of a candidate vaccine comprising the major outer membrane protein (MOMP) multiepitope of Ct delivered with the human papillomavirus (HPV) major capsid protein L1 as a vehicle with adjuvant properties, in a murine model of chlamydial genital infection. A recombinant plasmid pcDNA3.1(+) containing mammalian codon-optimization HPV6b L1 gene and Ct MOMP multiepitope was constructed. The Ct MOMP multiepitope containing T- and B-cell epitope-rich peptides was inserted into C-terminal of HPV6b L1-coding sequence. The constructed plasmid after verified by enzyme restriction assay and DNA sequencing was transfected into COS-7 cells. Expression of the chimeric gene in COS-7 cells was confirmed by RT-PCR, Western blot analysis and immunofluorescence assay. Results revealed successful expression of the chimeric HPV6b L1/Ct MOMP multiepitope gene both at the mRNA and protein levels in transfected COS-7 cells. Intramuscular (IM) administration in mice was able to elicit not only antibodies against Ct MOMP, but also Th1 and cytotoxic T lymphocyte activity against the Ct MOMP epitopes. In addition, recipients of IM immunization of HPV6b L1/Ct MOMP multiepitope were highly resistant to infection. Altogether, the results suggested that IM delivery of HPV6b L1-MOMP multiepitope may be a suitable vaccine regimen potentially capable of inducing protective mucosal immunity against Ct infection.

摘要

有效的疫苗接种是控制人类生殖道沙眼衣原体(Ct)感染的最长期有效措施。目前 Ct 疫苗开发的挑战是开发一种有效的递送载体,以诱导高水平的黏膜 T 细胞和补体 B 细胞反应。我们在沙眼衣原体生殖道感染的小鼠模型中评估了由 Ct 主要外膜蛋白(MOMP)多表位与具有佐剂特性的人乳头瘤病毒(HPV)主要衣壳蛋白 L1 组成的候选疫苗的免疫原性和疗效。构建了含有哺乳动物密码子优化 HPV6b L1 基因和 Ct MOMP 多表位的重组质粒 pcDNA3.1(+)。将含有 T 细胞和 B 细胞表位丰富肽的 Ct MOMP 多表位插入 HPV6b L1 编码序列的 C 末端。构建的质粒经酶切鉴定和 DNA 测序验证后,转染 COS-7 细胞。通过 RT-PCR、Western blot 分析和免疫荧光分析证实了嵌合基因在 COS-7 细胞中的表达。结果表明,在转染的 COS-7 细胞中,该嵌合 HPV6b L1/Ct MOMP 多表位基因在 mRNA 和蛋白质水平上均成功表达。在小鼠中肌内(IM)给药不仅能诱导针对 Ct MOMP 的抗体,还能诱导针对 Ct MOMP 表位的 Th1 和细胞毒性 T 淋巴细胞活性。此外,接受 HPV6b L1/Ct MOMP 多表位肌内免疫接种的受者对感染具有高度抵抗力。总之,这些结果表明,HPV6b L1-MOMP 多表位的 IM 递送可能是一种有潜力诱导针对 Ct 感染的保护性黏膜免疫的合适疫苗方案。

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