Generation of Robust Resident Memory T cells in Barrier Tissues through Skin Vaccination

通过皮肤疫苗接种在屏障组织中生成强大的常驻记忆 T 细胞

• 批准号: 10064958
• 负责人: Rachael Ann Clark
• 金额: $ 93.2万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-14 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10064958
• 关键词: Anatomy; Animal Model; Antigens; Asthma; Atopic Dermatitis; Binding; Biology; Blood; CD36 gene; Cell surface; Cells; Collaborations; Collection; Communicable Diseases; Data; Dependence; Evolution; FABP4 gene; Funding; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Generations; Genes; Goals; High-Throughput Nucleotide Sequencing; Homing; Human; Immune response; Immunity; Immunization; Immunization Programs; Immunize; Immunobiology; Individual; Infection; Infectious Agent; Influenza; Intramuscular; Lipase; Lipids; Lobectomy; Low Density Lipoprotein Receptor; Lung; Mediating; Medicine; Memory; Mesentery; Metabolism; Modified Vaccinia Virus Ankara; Molecular Analysis; Names; Nature; Nonesterified Fatty Acids; Organ; Patients; Peripheral; Phenotype; Play; Population; Poxviridae; Proliferating; Publishing; Recording of previous events; Research Personnel; Role; Route; Skin; Skin Tissue; Smallpox; Structure of parenchyma of lung; Surface; T memory cell; T-Cell Activation; T-Lymphocyte; T-cell receptor repertoire; TRB@ gene cluster; Testing; Thoracic Surgical Procedures; Time; Tissues; Translating; Translational Research; Tuberculin Test; United States National Institutes of Health; Vaccination; Vaccines; Vaccinia virus; Variola minor; Virus; Work; World Health Organization; cell mediated immune response; draining lymph node; fatty acid metabolism; fatty acid-binding proteins; human tissue; imprint; innovation; insight; lipid metabolism; mouse model; novel; novel strategies; overexpression; pathogen; pathology imaging; respiratory pathogen; skin vaccination; subcutaneous; translational medicine; translational study; uptake; vaccine development; vector

This R01 application proposes to continue work funded for the past five years by a Transformative R01 (“Vaccination to generate tissue resident memory T cells”) from the Office of the NIH Director (TR01’s cannot be renewed as such). The previous funding period allowed Drs. Kupper and Clark to make fundamental insights into immunobiology of TRM in skin and other tissues, in both mouse models and humans. Taken together, the body of work published by both co-PI’s over the past five years validates the importance of TRM in protective immunity, is summarized in commissioned reviews published in 2015 by Dr. Clark (in Science Translational Medicine) and Dr. Kupper (in Nature Medicine). The present application builds on exciting preliminary data showing that tissue specific imprinting occurs very early after T cell activation, and that the gene expression profile of T cells activated in skin draining and lung draining lymph nodes (LN) is surprisingly similar (but very different from gut draining LN). This may explain why immunization with a replication deficient poxvirus (MVA) vector through epidermal disruption (formerly “skin scarification”, or ed/ss) generates not only skin TRM but also lung TRM with very high efficiency. This suggests that ed/ss skin immunization may be a highly effective way of generating protective immunity in the lung to pulmonary pathogens like influenza. This hypothesis will be tested in Aim 1. Conventional vaccination routes (intradermal, subcutaneous, and intramuscular) were much less efficient at generating TRM in both skin and lung. Remarkably, the gene expression profile of T cells responding to the same antigen delivered by these different routes was very different, with ep/ss and i.m. being the most different. These differences in gene expression were maintained even 30 days later in splenic TEM, and were manifested by a strikingly different capacity to generate TRM. Some of the greatest differences in gene expression between TRM and TCM are in genes relevant to lipid metabolism (fatty acid binding proteins/FABP, lipases, and CD36/LDL receptor); TRM’s generated from T cells deficient in FABP4 and 5 did not persist in skin. The hypothesis that the ep/ss mode of immunization most efficiently generates TRM that are dependent on fatty acid metabolism for survival in peripheral tissues will be tested in Aim 2. Finally, in Aim 3, related hypotheses will be tested in translational studies using human tissue, with blood, skin, and lung tissue being obtained from thoracic surgery patients undergoing lobectomies. We predict, and will test for, overlap (at the level of CDR3 sequence) in the T cell repertoire of skin and lung by high throughput sequencing of the TCRB gene, and will directly compare human skin and lung TRM by gene expression, cell surface phenotype, and function. The previously unappreciated commonalities in skin and lung homing T cells may explain phenomena from the tuberculin skin test to the “atopic march”, where atopic dermatitis is followed by asthma, and has important implications for human vaccine development against pathogens that enter the host through lung or skin.

此 R01 申请建议继续过去五年由变革性 R01 资助的工作 （“疫苗接种以生成组织驻留记忆 T 细胞”）来自 NIH 主任办公室（TR01 不能 如此更新）。之前的资助期允许博士。库珀和克拉克奠定了基础 深入了解 TRM 在小鼠模型和人类皮肤和其他组织中的免疫生物学。采取 两位联合 PI 在过去五年中共同发表的著作证实了 TRM 在 Clark 博士在 2015 年发表的委托评论中总结了保护性免疫（《Science》 转化医学）和库珀博士（自然医学）。本应用程序建立在令人兴奋的基础上 初步数据表明，T 细胞激活后很早就发生了组织特异性印记，并且 皮肤引流淋巴结和肺引流淋巴结 (LN) 中激活的 T 细胞的基因表达谱令人惊讶 类似（但与肠道引流液氮有很大不同）。这可以解释为什么复制缺陷免疫 痘病毒（MVA）载体通过表皮破坏（以前的“皮肤划痕”，或 ed/ss）不仅产生 皮肤 TRM 以及肺 TRM 都具有非常高的效率。这表明 ed/ss 皮肤免疫可能是 在肺部产生针对流感等肺部病原体的保护性免疫力的高效方法。这 假设将在目标 1 中进行检验。传统的疫苗接种途径（皮内、皮下和 肌内注射）在皮肤和肺部产生 TRM 的效率要低得多。值得注意的是，该基因 T 细胞对通过这些不同途径递送的相同抗原作出反应的表达谱非常 不同的是，ep/ss 和 i.m.是最不同的。这些基因表达的差异得以维持 甚至在 30 天后的脾脏 TEM 中，也表现出产生 TRM 的能力截然不同。 TRM 和 TCM 之间基因表达的一些最大差异在于与脂质相关的基因 代谢（脂肪酸结合蛋白/FABP、脂肪酶和 CD36/LDL 受体）； TRM 由 T 细胞产生 FABP4 和 5 缺陷不会在皮肤中持续存在。假设 ep/ss 免疫模式最有效 有效地产生依赖于脂肪酸代谢在外周组织中生存的TRM 在目标 2 中进行测试。最后，在目标 3 中，相关假设将在使用人体组织的转化研究中进行测试， 血液、皮肤和肺组织取自接受肺叶切除术的胸外科患者。我们 预测并测试皮肤和肺 T 细胞库中的重叠（在 CDR3 序列水平） 对TCRB基因进行高通量测序，并通过基因直接比较人体皮肤和肺TRM 表达、细胞表面表型和功能。以前未被认识到的皮肤和皮肤的共性 肺归巢 T 细胞可以解释从结核菌素皮试到“特应性进行曲”的现象，其中特应性 皮炎之后是哮喘，这对人类疫苗的开发具有重要意义 通过肺部或皮肤进入宿主的病原体。