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一种用于病毒载体疫苗对山羊阴道黏膜转导进行体外监测的简单通用方法。

A Simple and Versatile Method for Ex Vivo Monitoring of Goat Vaginal Mucosa Transduction by Viral Vector Vaccines.

作者信息

Minesso Sergio, Odigie Amienwanlen Eugene, Franceschi Valentina, Cotti Camilla, Cavirani Sandro, Tempesta Maria, Donofrio Gaetano

机构信息

Department of Veterinary Science, University of Parma, 43126 Parma, Italy.

Department of Veterinary Medicine, University of Bari, 70010 Valenzano, Italy.

出版信息

Vaccines (Basel). 2024 Jul 29;12(8):851. doi: 10.3390/vaccines12080851.

Abstract

Goat may represent a valid large animal model for human pathogens and new vaccines testing. Appropriate vaccine administration is a critical component of a successful immunization program. The wrong route of administration may reduce the efficacy of the vaccine, whereas the proper administration strategy can enhance it. Viral vectors have been employed successfully for goat and sheep immunization; however, no data concerning the vaginal route are available. A viral vector's ability to transduce the site of inoculation is of primary interest. In this study, a fast and reliable ex vivo assay for testing the transduction capability of an Ad5-based vector when intravaginally administered was developed. An Ad5 vector delivering an expression cassette with a bicistronic reporter gene, Ad5-CMV-turboGFP-IRES-Luc2, was constructed. We demonstrated Ad5-CMV-turboGFP-IRES-Luc2's ability to transduce caprine vaginal mucosa by ex vivo bioluminescent imaging (BLI) employing a simple CCD camera apparatus for chemiluminescence western immunoblotting. These data, though simple, provide valuable insights into developing a vaginal immunization strategy using a viral vector-based vaccine to protect against pathogens causing genital diseases.

摘要

山羊可能是用于人类病原体和新型疫苗测试的有效大型动物模型。合适的疫苗接种途径是成功免疫计划的关键组成部分。错误的接种途径可能会降低疫苗效力,而正确的接种策略则可增强效力。病毒载体已成功用于山羊和绵羊的免疫接种;然而,尚无关于阴道途径的数据。病毒载体转导接种部位的能力是主要关注点。在本研究中,开发了一种快速可靠的体外试验,用于测试基于Ad5的载体经阴道给药时的转导能力。构建了一种携带双顺反子报告基因表达盒的Ad5载体,即Ad5-CMV-turboGFP-IRES-Luc2。我们通过使用简单的化学发光western免疫印迹电荷耦合器件(CCD)摄像装置的体外生物发光成像(BLI),证明了Ad5-CMV-turboGFP-IRES-Luc2转导山羊阴道黏膜的能力。这些数据虽然简单,但为开发基于病毒载体疫苗的阴道免疫策略以预防引起生殖器疾病的病原体提供了有价值的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb4f/11359855/bdd837b1c2e7/vaccines-12-00851-g001.jpg

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