Topical DNA immunization by modifying the hair follicle cycle

通过改变毛囊周期进行局部 DNA 免疫

• 批准号: 7533889
• 负责人: ZHENGRONG CUI
• 金额: $ 35.44万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7533889
• 关键词: Adoption; Affect; Animals; Anthrax Vaccines; Anthrax disease; Antigens; Area; Bacillus anthracis; Biology; Cells; Chemicals; Cholera Toxin; Clinical; Clinical Trials; Communicable Diseases; DNA; DNA Vaccines; Data; Dendritic Cells; Dermal; Development; Disruption; Drug Formulations; Epidermis; Foundations; Future; Gene Expression; Generations; Genes; Goals; Growth; Hair follicle structure; Health Personnel; Histopathology; Human; Immune; Immune response; Immune system; Immunization; Infection; Inflammation; Invasive; Mass Immunization; Modality; Modeling; Outcome; Penetration; Personal Satisfaction; Plasmids; Play; Proteins; Public Health; Reporter Genes; Reporting; Research Personnel; Rest; Role; Self Administration; Skin; Staging; Stratum corneum; Surface; System; Technology; Testing; Time; Topical application; Training; Transdermal substance administration; Transfection; Vaccine Adjuvant; Vaccines; Validation; Work; anthrax protective factor; base; early onset; experience; improved; innovation; interest; novel; novel strategies; pathogen; plasmid DNA; prevent; sound; success; vaccine delivery; vaccine development

DESCRIPTION (provided by applicant): The skin acts as a pathogen barrier, and the skin immune system is well-equipped for the generation of effective anti-infection immune responses. The induction of immune responses by applying a DNA vaccine topically onto the skin is an attractive immunization approach. Unfortunately, the immune responses induced using this approach is generally weak, evidently due to the difficulty for the macromolecular plasmid DNA to penetrate through the skin stratum corneum to reach the viable skin cells to allow the antigen gene(s) encoded by the plasmid to be expressed. Data from several recent studies pointed out that the hair follicles are likely to be the portal of entrance for topically applied DNA. When plasmid DNA was applied onto the skin, the expression of the gene(s) encoded by the plasmid was largely confined in the hair follicles and only occasionally in other cells in the epidermis. Moreover, the finding that the presence of normal hair follicles in the skin was required for a topical DNA vaccine to induce specific immune responses further suggested the hair follicles as the portal of entrance for topically applied plasmid DNA. Normal hair follicles cycle between growth (anagen) and resting stages. It was reported that hair follicles in the growth stage are open for penetration by foreign objects on the skin surface, whereas those in the resting stage are closed. In fact, the expression of a reporter gene applied topically onto the skin was significantly enhanced when the hair follicles in the application area were induced into anagen onset (early growth) stage. Based on all these findings, we hypothesized that the immune responses induced by a topical DNA vaccine would be enhanced when the hair follicles in the application area were induced into anagen onset stage. Using a plasmid that encodes the protective antigen (PA) protein of Bacillus anthracis as a model DNA vaccine, we have generated some promising data that were strongly supportive of this hypothesis, indicating that topical immunization by applying a DNA vaccine onto a skin area where the hair follicles are in, or induced into, anagen onset stage represents a viable immunization modality. However, before validating this approach in humans, we propose to further evaluate it by carrying out the following four specific aims: (i) to elucidate the mechanisms responsible for the enhancement of the immune response induced by a topical DNA vaccine when the hair follicles in the application area are induced into anagen (onset) stage (aim # 1), (ii) to identify factors that affect the immune responses induced by a topical DNA vaccine when the hair follicles in the application area are in, or induced into, anagen stage (aim # 2), (iii) to evaluate the extent to which the immune responses induced by a DNA vaccine using this approach will protect the animals against a lethal pathogen challenge (aim # 3), and to elucidate the mechanisms of immune induction by a DNA vaccine applied topically onto a skin area where the hair follicles are in anagen stage (aim # 4). We have assembled a team of researchers with extensive experience in vaccine development, transdermal drug delivery, skin biology, and histopathology to safeguard the successful completion of the proposed studies. The outcomes will lay a sound scientific foundation for us to improve the efficacy of topical DNA vaccines in future trials. PUBLIC HEALTH RELEVANCE: The successful completion of the proposed studies will lay a sound foundation for the development and validation of this novel topical DNA immunization approach in future (clinical) trials by modifying the hair follicle cycle. Such an efficacious non-invasive topical DNA immunization modality is expected to be amenable for mass immunization to prevent infectious diseases in humans and animals.

描述（由申请人提供）：皮肤充当病原体屏障，皮肤免疫系统能够产生有效的抗感染免疫反应。通过将 DNA 疫苗局部涂抹在皮肤上来诱导免疫反应是一种有吸引力的免疫方法。不幸的是，使用这种方法诱导的免疫反应通常较弱，显然是由于大分子质粒DNA难以穿透皮肤角质层到达有活力的皮肤细胞，从而使质粒编码的抗原基因被释放。表示。最近几项研究的数据指出，毛囊很可能是局部应用 DNA 的入口。当质粒DNA被应用到皮肤上时，由质粒编码的基因的表达主要局限于毛囊中，并且偶尔在表皮的其他细胞中表达。此外，外用 DNA 疫苗需要皮肤中存在正常毛囊才能诱导特异性免疫反应，这一发现进一步表明毛囊是外用质粒 DNA 的入口门户。正常毛囊在生长（生长期）和静止阶段之间循环。据报道，生长期的毛囊是开放的，可以被皮肤表面的异物穿透，而休止期的毛囊是封闭的。事实上，当应用区域的毛囊被诱导进入生长期开始（早期生长）阶段时，局部应用到皮肤上的报告基因的表达显着增强。基于所有这些发现，我们假设当应用区域的毛囊被诱导进入生长期开始阶段时，局部 DNA 疫苗诱导的免疫反应将会增强。使用编码炭疽杆菌保护性抗原（PA）蛋白的质粒作为模型 DNA 疫苗，我们产生了一些有希望的数据，这些数据强烈支持这一假设，表明通过将 DNA 疫苗应用于皮肤区域进行局部免疫，其中毛囊处于或诱导进入生长期开始阶段代表了可行的免疫方式。然而，在人体验证这种方法之前，我们建议通过执行以下四个具体目标来进一步评估它：(i) 阐明当毛囊处于毛囊中时，局部 DNA 疫苗诱导免疫反应增强的机制。应用区域被诱导进入生长期（开始）阶段（目标＃1），（ii）确定当应用区域中的毛囊处于或诱导进入生长期时影响局部DNA疫苗诱导的免疫反应的因素阶段（目标＃2），（iii）评估使用这种方法的DNA疫苗诱导的免疫反应将在多大程度上保护动物免受致命病原体的攻击（目标＃3），并阐明免疫诱导机制通过将 DNA 疫苗局部涂抹到毛囊处于生长期阶段的皮肤区域（目标#4）。我们组建了一支在疫苗开发、透皮给药、皮肤生物学和组织病理学方面拥有丰富经验的研究人员团队，以保障拟议研究的成功完成。研究结果将为我们在未来的试验中提高外用DNA疫苗的功效奠定良好的科学基础。公共健康相关性：拟议研究的成功完成将为通过改变毛囊周期在未来（临床）试验中开发和验证这种新型局部 DNA 免疫方法奠定良好的基础。这种有效的非侵入性局部DNA免疫方式有望适用于大规模免疫，以预防人类和动物的传染病。