Use of Adjuvants to Improve Skin Genetic Immunizations

使用佐剂改善皮肤遗传免疫

• 批准号: 6858587
• 负责人: Adriana T Larregina
• 金额: $ 27.06万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-18 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6858587
• 关键词: T lymphocyte; antigen presentation; antineoplastics; cell migration; cell population study; clinical research; dendritic cells; gene delivery system; genetically modified animals; human tissue; immune response; immunomodulators; laboratory mouse; melanoma; neoplasm /cancer genetics; neoplasm /cancer vaccine; quinoline analog; tissue /cell culture; transfection; tumor antigens; vector vaccine

DESCRIPTION (provided by applicant): Melanomas are one the most aggressive tumors with increasing incidence and mortality. Although melanomas are immunogenic tumors that could be potentially eliminated by immune-mediated spontaneous regression, this phenomenon is extremely rare. More frequently, melanomas are not recognized by the immune system due to the absence of tumor-specific antigens (Ags) or development of a state of tumor-specific tolerance. Immunization strategies against melanoma by means of DNA vaccines are promising therapeutic tools based on the induction of tumor-specific CD4+ T helper 1 (Th1) lymphocytes and CD8+ cytotoxic T cells (CTLs). Due to its easy accessibility and intrinsic immune system, the skin is an optimal site for genetic immunization and the gene gun (GO is an excellent approach to induce high expression of transgenic Ags in the skin. Professional Ag processing and presenting cells of the skin [cutaneous dendritic cells (DCs): epidermal Langerhans cells and dermal DCs] can be transfected in situ by GG-immunization, or alternatively, they may internalize transgenic proteins from neighboring skin cells. Therefore, through direct of indirect transfection of skin DCs by the GG, it is possible to engineer in vivo the skin immune system. The presence of appropriate immuno-stimulatory signals during the early stages of DCs activation in peripheral tissues (i.e. skin) "instructs" DCs to fully mature and to induce an effective anti-tumor Th1-biased immune response. However, although the GG induces anti-tumor CD8+ CTLs, it is still controversial whether the GG is able to generate an effective CD4+ Th1-biased immune response (required to maintain efficient cellular anti-tumor immunity) and it rather induces Th2 cells. The hypothesis of this proposal is that the use of the GG to express transgenic melanoma Ags in the skin in the presence of the Th1-driving adjuvants substance P neurokinin 1 receptor agonist (SP-NK1a) and Imiquimod (to mature and to Th1-polarize skin DCs at the site of GG-immunization) will generate fully activated skin DCs with ability to stimulate efficiently tumor Ag-specific T cells. Skin-derived DCs generated under these conditions will bias the differentiation of naive tumor-specific T cells into Th1 cells and promote development of CTLs, generating an effective and long-lasting anti-tumor immune response. To test the hypothesis, we propose the following specific aims: Aim 1: To investigate the function of DCs migrated from skin transfected with GG in the presence of Th1-driving adjuvants Imiquimod and SP-NK1a. Aim 2: To analyze in vivo the role of Th1-driving adjuvants in the outcome of the immune responses induced by GG genetic immunization of skin, and Aim 3: To analyze the capacity of the Th1-driving adjuvants to polarize different subpopulations of skin migratory DCs from human skin transfected with the GG.

描述（由申请人提供）：黑色素瘤是最具侵袭性的肿瘤之一，发病率和死亡率不断增加。虽然黑色素瘤是免疫原性肿瘤，可以通过免疫介导的自发消退来消除，但这种现象非常罕见。更常见的是，由于缺乏肿瘤特异性抗原（Ag）或肿瘤特异性耐受状态的发展，黑素瘤不被免疫系统识别。通过DNA疫苗对黑色素瘤的免疫策略是基于诱导肿瘤特异性CD 4+辅助性T 1（Th 1）淋巴细胞和CD 8+细胞毒性T细胞（CTL）的有前途的治疗工具。由于其易于接近和固有的免疫系统，皮肤是基因免疫的最佳位点，并且基因枪（GO）是诱导转基因Ag在皮肤中高表达的极好方法。皮肤的专业Ag加工和呈递细胞[皮肤树突状细胞（DC）：表皮朗格汉斯细胞和真皮DC]可以通过GG免疫原位转染，或者可选地，它们可以内化来自邻近皮肤细胞的转基因蛋白。因此，通过GG直接或间接转染皮肤DC，可以在体内工程化皮肤免疫系统。在外周组织（即皮肤）中DC活化的早期阶段，适当的免疫刺激信号的存在“指示”DC完全成熟并诱导有效的抗肿瘤Th 1偏向的免疫应答。然而，尽管GG诱导抗肿瘤CD 8 + CTL，但GG是否能够产生有效的CD 4 + Th 1-偏向的免疫应答（维持有效的细胞抗肿瘤免疫所需的），而不是诱导Th 2细胞，仍然存在争议。该建议的假设是，在Th 1驱动佐剂P物质神经激肽1受体激动剂（SP-NK 1a）和咪喹莫特（使GG免疫部位的皮肤DC成熟并使其成为Th 1-活化的皮肤DC）存在下，使用GG在皮肤中表达转基因黑色素瘤Ag将产生完全活化的皮肤DC，其具有有效刺激肿瘤Ag特异性T细胞的能力。在这些条件下产生的皮肤来源的DC将使初始肿瘤特异性T细胞偏向于分化为Th 1细胞，并促进CTL的发育，产生有效且持久的抗肿瘤免疫应答。为了验证这一假设，我们提出了以下具体目标：目的1：研究在Th 1驱动佐剂咪喹莫特和SP-NK 1a存在下，转染GG的皮肤中迁移的DC的功能。目标二：分析Th 1驱动佐剂在GG基因免疫皮肤诱导的免疫应答中的作用。目的3：分析Th 1驱动佐剂对GG基因转染的人皮肤迁移性DC不同亚群的诱导能力。