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应用程序建议继续过去五年由变革性R01资助的工作 (“接种疫苗以产生组织常驻记忆T细胞”)，来自美国国立卫生研究院院长办公室(TR01‘S不能 按此方式续期)。上一个资助期允许Kper博士和Clark博士 对皮肤和其他组织中TRM免疫生物学的洞察，无论是在小鼠模型中还是在人类中。已被占用 在过去的五年里，这两家公司共同出版的著作证实了TRM在 保护性免疫在克拉克博士2015年发表的委托评论中进行了总结(发表在《科学》杂志上 转化医学)和库珀博士(自然医学)。目前的应用建立在令人兴奋的基础上 初步数据显示，组织特异性印记在T细胞激活后很早就发生，而且 皮肤引流和肺引流淋巴结(LN)中活化的T细胞基因表达谱令人惊讶 相似(但与肠道引流LN非常不同)。这可能解释了为什么复制缺陷的免疫 痘病毒(MVA)载体通过表皮破裂(以前称为皮肤划痕，或ed/ss)不仅产生 皮肤TRM和肺TRM也具有很高的效率。这表明ED/SS皮肤免疫可能是一种 在肺部对流感等肺部病原体产生保护性免疫的高效方式。这 假设将在目标1中得到检验。常规接种途径(皮内、皮下和 肌肉内)在皮肤和肺部产生TRM的效率要低得多。值得注意的是，这种基因 通过这些不同途径传递的T细胞对同一抗原的反应表达谱非常 不同的是，EP/SS和IM。是最不同的。这些基因表达的差异保持不变 即使在30天后，在脾的透射电子显微镜下，显示出显著不同的产生TRM的能力。 TRM和中医在基因表达上的一些最大差异是与脂肪相关的基因 代谢(脂肪酸结合蛋白/FABP、脂肪酶和CD36/低密度脂蛋白受体)；T细胞产生的TRM FABP4和FABP5缺乏在皮肤中不能持续。关于EP/SS免疫模式的假设 在周围组织中有效地产生依赖于脂肪酸代谢生存的TRM将是 最后，在目标3中，相关假设将在使用人体组织的翻译研究中得到验证， 血液、皮肤和肺组织取自接受肺叶切除术的胸部手术患者。我们 通过以下方式预测并将测试皮肤和肺的T细胞库中的重叠(CDR3序列水平) TCRB基因的高通量测序，并将直接比较人皮肤和肺的TRM基因 表达、细胞表面表型和功能。以前没有意识到的皮肤和皮肤上的共性 肺归巢T细胞可以解释从结核菌素皮肤试验到特应性进行曲的现象 皮炎紧随哮喘之后，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。