Use of Adjuvants to Improve Skin Genetic Immunizations

使用佐剂改善皮肤遗传免疫

• 批准号: 7409594
• 负责人: Adriana T Larregina
• 金额: $ 25.63万
• 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 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7409594
• 关键词: Adjuvant; Agonist; Apoptosis; Automobile Driving; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell Adhesion Molecules; Cell Communication; Cell surface; Cells; Cellular Immunity; Chickens; Condition; Cutaneous; Cutaneous Administration; Cytotoxic T-Lymphocytes; DNA; DNA Vaccines; Dendritic Cells; Dendritic cell activation; Dermal; Development; Disease regression; Early Diagnosis; Engineering; Future; Generations; Human; Humoral Immunities; Imiquimod; Immune; Immune response; Immune system; Immunity; Immunization; In Situ; Incidence; Inflammatory; Inflammatory Response; Interferons; Interleukin-10; Interleukin-12; Interleukin-2; Interleukin-4; Interleukin-5; Langerhans cell; Lymphocyte; Lymphoid; Lymphoid Tissue; Maintenance; Mediating; Mediator of activation protein; Memory; Modeling; Mus; Natural Killer Cells; Operative Surgical Procedures; Organ; Outcome; Pattern; Peptides; Peripheral; Principal Investigator; Process; Proteins; Resistance; Role; Secondary to; Self Tolerance; Signal Transduction; Site; Skin; Skin Neoplasms; Staging; Substance P; Substance P Receptor; Surface; T-Lymphocyte; Testing; Th2 Cells; Therapeutic; Therapeutic Effect; Time; Tissues; Transfection; Transgenes; Transgenic Organisms; Tumor Antigens; Tumor Immunity; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Tumor-Associated Process; Tumor-Derived; UV induced; anergy; base; chemotherapy; cytokine; gene gun; genetic immunization strategies; human TNF protein; immunogenic; immunological synapse; improved; in vivo; irradiation; keratinocyte; killings; lymph nodes; macrophage; melanoma; mortality; neoplastic cell; programs; response; tool; tumor

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，其发病率和死亡率不断增加。尽管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，但这种现象极为罕见。更常见的是，由于缺乏肿瘤特异性抗原（Ag）或形成肿瘤特异性耐受状态，免疫系统无法识别黑色素瘤。通过 DNA 疫苗对抗黑色素瘤的免疫策略是基于肿瘤特异性 CD4+ T 辅助 1 (Th1) 淋巴细胞和 CD8+ 细胞毒性 T 细胞 (CTL) 诱导的有前景的治疗工具。由于其易于接近和固有的免疫系统，皮肤是基因免疫和基因枪的最佳部位（GO是诱导皮肤中转基因Ag高表达的绝佳方法。专业的Ag处理和呈递皮肤细胞[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， 它们可能会内化邻近皮肤细胞的转基因蛋白质。因此，通过GG直接或间接转染皮肤DC，可以在体内改造皮肤免疫系统。在外周组织（即皮肤）DCs激活的早期阶段，适当的免疫刺激信号的存在“指示”DCs完全成熟并诱导有效的抗肿瘤Th1偏向性 免疫反应。然而，尽管GG诱导抗肿瘤CD8+ CTL，但GG是否能够产生有效的CD4+ Th1偏向免疫反应（维持有效的细胞抗肿瘤免疫所需）并且诱导Th2细胞仍存在争议。该提案的假设是，在存在 GG 的情况下，使用 GG 在皮肤中表达转基因黑色素瘤 Ag Th1 驱动佐剂 P 神经激肽 1 受体激动剂 (SP-NK1a) 和咪喹莫特（使 GG 免疫部位的皮肤 DC 成熟并实现 Th1 极化）将产生完全活化的皮肤 DC，能够有效刺激肿瘤 Ag 特异性 T 细胞。在这些条件下产生的皮肤来源的 DC 将使幼稚肿瘤特异性 T 细胞分化为 Th1 细胞并促进 CTL 的发育，产生有效且持久的抗肿瘤免疫反应。为了检验这一假设，我们提出以下具体目标： 目标 1：研究在 Th1 驱动佐剂咪喹莫特和 SP-NK1a 存在的情况下，从 GG 转染的皮肤迁移的 DC 的功能。目标 2：体内分析 Th1 驱动佐剂在结果中的作用 皮肤GG基因免疫诱导的免疫反应，目标3：分析Th1驱动佐剂极化来自转染GG的人皮肤的皮肤迁移DC的不同亚群的能力。