Exploring mechanisms that govern immune homeostasis in skin

我们身体最大的器官是皮肤，它是抵御环境压力的重要屏障，比如温度变化、阳光、过敏原、毒素和微生物。这个复杂的器官由许多细胞类型组成，包括上皮细胞、成纤维细胞、神经元和脉管系统，这些细胞共同作用以确保其功能。免疫系统的细胞已经成为皮肤生物学许多方面的协调者，共同加强抵御外部威胁的屏障。免疫细胞的组成、功能和激活状态不断受到皮肤结构、微生物和炎症刺激的调节。反过来，皮肤常驻和招募的免疫细胞与组织实质积极对话，调节共生细菌，限制微生物入侵，直接炎症和修复。以前的研究主要集中在朗格汉斯细胞（LC）的功能和生物学上，这是表皮中独特的抗原呈递细胞，其在体内的功能自发现以来一直不清楚。我们证明LCs具有一种独特的机制，可以获得突破皮肤最外层屏障角质层的外来抗原，但仍在表皮紧密连接之外（Kubo等人，J Exp Med 2009）。这使我们假设LCs调查的共生菌在某些情况下也可能是致病的。为了解决这个问题，我们将金黄色葡萄球菌衍生毒素接种到完整的皮肤上。虽然这不会引起皮肤炎症，但我们发现LCs通过完整的紧密连接屏障获得毒素，并诱导产生中和抗体，保护实验性葡萄球菌烫伤皮肤综合征小鼠免受毒素的全身攻击（Ouchi等人，J Exp Med 2011）。我们进一步证明，LCs吸收表皮自身抗原，诱导抑制自身免疫性皮肤病的调节性T细胞的扩增（Kitashima等人，eBioMedicine, 2018）。总的来说，这些研究确定了LCs在宿主防御和自身免疫中的关键作用。导致当前组织免疫串扰主题的一项重要成就是阐明了毛囊（哺乳动物的定义特征之一）如何维持LC网络。我们发现毛囊是具有免疫活性的结构，在感知机械应力时，毛囊产生趋化因子来吸引LC前体，并作为其重新繁殖到表皮的门户（Nagao等人，Nat Immunol 2012）。这一发现代表了组织特异性信号如何与免疫细胞沟通以维持免疫稳态的新概念。我们最近扩展了上述发现，证明毛囊产生细胞因子，使表皮中的记忆T细胞持续存在。重要的是，当常驻记忆T细胞恶性转化为淋巴瘤时，它们仍然依赖毛囊来源的细胞因子（Adachi et al, Nat Med 2015）。我们还通过建立一个ADAM17 cKO特应性皮炎（AD）小鼠模型，研究了疾病环境中宿主-微生物的相互作用，该模型自发发展为与生态失调（细菌菌群失衡）相关的湿疹性皮肤炎症，主要是金黄色葡萄球菌，这是人类AD的一个特征。金黄色葡萄球菌在AD皮肤上的定植是否会导致湿疹性炎症或仅仅是慢性炎症的结果一直存在争议。我们在AD小鼠模型中确定金黄色葡萄球菌是湿疹形成的关键组成部分，为长期存在的临床问题提供了答案（Kobayashi等人，Immunity 2015）。通过对ADAM17 cKO小鼠皮肤的大量rna测序分析，我们发现这些小鼠的转录组与人类AD的转录组相似（Woodring等人，J Invest Dermatol, 2018）。我们进一步证明，表皮的先天淋巴样细胞（ILC）依赖毛囊来源的细胞因子和趋化因子在皮脂腺附近持续存在和定位。在那里，ilc调节皮脂腺的功能，以调节皮肤表面微生物的平衡（Kobayashi等人，Cell, 2019）。在临床方面，我对药物过敏有长期的临床兴趣，在我过渡到NIH之前，我曾担任日本严重皮肤不良反应委员会（JSCAR）的成员，这是一个政府资助的共识小组。在此期间，我们进行了一项临床研究，以了解药物性超敏反应综合征（DiHS，也称为嗜酸性粒细胞增多和全身症状的药物反应或DRESS）后发生的后遗症，并确定这种药物超敏反应会导致自身免疫性疾病的发生（Kano et al, J Dermatol. 2015）。我们还确定了高剂量免疫球蛋白作为Stevens-Johnson综合征和中毒性表皮坏死松解症的替代疗法的疗效，这些疾病中常驻记忆T细胞可能发挥重要作用（Aihara等，J Dermatol. 2015）。此外，我们确定万古霉素诱导的线性IgA大疱性皮肤病的发生是万古霉素与预先存在的IgA储存库的直接相互作用，使它们对皮肤基底膜的关键成分VII型胶原产生反应，表现为自身免疫性水疱病（Yamagami等人，J Invest Dermatol, 2018）。我们已经成功地应用单细胞RNA测序技术在难治性DIHS/DRESS患者中确定了治疗靶点，为我们如何加深对复杂疾病病理生理学的理解以及个性化医疗打开了大门（Kim et al, Nat Med, 2020）。最后，我们发现中性粒细胞在Stevens-Johnson综合征和中毒性表皮坏死松解的早期阶段触发炎症（Kinoshita et al, Sci Transl Med, 2021）。在NIAMS皮肤白细胞生物学部分，我们进一步探索组织免疫和宿主-微生物在体内平衡和炎症过程中相互作用的基本机制。这些过程涉及先天免疫细胞和适应性免疫细胞，我们假设上皮或间质室之间的串扰改变参与了皮肤病的病理生理。我们采用临床相关的模型，包括AD和脱发的小鼠模型，以扩大我们对组织免疫串音的基础知识，并为更好地理解皮肤免疫和炎症性皮肤病奠定基础。

Our body's largest organ, the skin, is a vital barrier against environmental pressures such as thermal changes, sunlight, allergens, toxins, and microbes. This complex organ is composed of an array of many cell types including epithelia, fibroblasts, neurons, and vasculature that act in unison to ensure its function. Cells of the immune system have emerged as orchestrators of many facets of skin biology that collectively serve to reinforce the barrier against external threats. The composition, function, and activation status of immune cells is constantly tuned by skin structures, microbial and inflammatory stimuli. In turn, skin-resident and recruited immune cells engage in an active dialogue with the tissue parenchyma to regulate commensal bacteria, limit microbial invasion and direct inflammation and repair. Previous research program primarily focused on the functions and biology of Langerhans cells (LC), antigen presenting cells unique in the epidermis, who's in vivo functions had remained unclear since their discovery. We demonstrated that LCs were equipped with a unique mechanism to gain access to foreign antigens that had breached the skin's outermost barrier, the stratum corneum, but that were still outside of epidermal tight junctions (Kubo et al, J Exp Med 2009). This led us to hypothesize that LCs survey for commensal bacteria which may also be pathogenic under certain circumstances. To address this, we inoculated Staphylococcus aureus-derived toxin onto intact skin. While this caused no skin inflammation, we found that LCs acquired the toxin through intact tight junction barriers and induced the production of neutralizing antibodies that protected mice from systemic challenge of the toxin in experimental Staphylococcal scalded skin syndrome (Ouchi et al, J Exp Med 2011). We further demonstrated that LCs took up an epidermal autoantigen to induce the expansion of regulatory T cells that suppressed autoimmune skin disease (Kitashima et al, eBioMedicine, 2018). These studies, in aggregate, established crucial roles of LCs during host defense and autoimmunity. An important accomplishment that led to the current theme of tissue-immune crosstalk was elucidating how the LC network is maintained by hair follicles, one of the defining features of mammals. We discovered that hair follicles are immunologically active structures that, upon sensing mechanical stress, produced chemokines to attract LC precursors and served as a gateway for their repopulation into epidermis (Nagao et al, Nat Immunol 2012). This finding represented a novel concept of how tissue-specific signals communicate with immune cells to maintain immunological homeostasis. We have recently expanded on the above findings by demonstrating that hair follicles produced cytokines that enabled the persistence of memory T cells in the epidermis. Importantly when resident memory T cells underwent malignant transformation to lymphoma, they remained dependent on hair follicle-derived cytokines (Adachi et al, Nat Med 2015). We also studied host-microbe interactions in a disease setting by generating an ADAM17 cKO mouse model of atopic dermatitis (AD) that spontaneously developed eczematous skin inflammation associated with dysbiosis (imbalance of the bacterial flora) that was predominated by S. aureus, a feature that recapitulates human AD. Whether S. aureus colonization on AD skin contributed to eczematous inflammation or was merely a result of chronic inflammation had been debated. We determined in our AD mouse model that S. aureus was a crucial component of eczema formation, providing an answer to a long-standing clinical question (Kobayashi et al, Immunity 2015). Using bulk RNA-seq analysis of skin from ADAM17 cKO mice we demonstrated that the transcriptome in these mice recapitulates that of human AD (Woodring et al, J Invest Dermatol, 2018). We further demonstrated that innate lymphoid cells (ILC) in the epidermis rely on hair follicle-derived cytokines and chemokines for persistence and localization near the sebaceous glands. There, the ILCs regulate the functions of sebaceous glands to tune the equilibrium of microbes that reside on skin surface (Kobayashi et al, Cell, 2019). On the clnical side, I have lonstanding clinical interest in drug hypersensitivity and have served as a member of the Japanese Severe Cutaneous Adverse Reactions (JSCAR) committee, a government-funded consensus group, prior to my transition to the NIH. During this period, we performed a clinical study to understand sequelae that occur after drug-induced hypersensitivity syndrome (DiHS, also known as drug reaction with eosinophilia and systemic symptoms or DRESS) and have established that this drug hypersensitivity leads to the onset of autoimmune diseases (Kano et al, J Dermatol. 2015). We also determined the efficacy of high-dose immunoglobulin as an alternative therapy in Stevens-Johnson syndrome and toxic epidermal necrolysis, diseases in which resident-memory T cells may play important roles (Aihara et al, J Dermatol. 2015). Furthermore, we determined that vancomycin-induced linear IgA bullous dermatosis occurs as a direct interaction of vancomycin with preexisting IgA reservoir that renders them reactive against type VII collagen, a critical component of the basal membrane in skin, manifesting as an autoimmune blistering disease (Yamagami et al, J Invest Dermatol, 2018). We have successfully applied the single-cell RNA sequencing technology to identify a therapeutic target in a treatment-refractory DIHS/DRESS patient, opening doors for how we can deepen our understanding of the complicated disease pathophysiology and also for personalized medicine (Kim et al, Nat Med, 2020). Finally, we have identified neutrophils to trigger inflammation in the early stages of Stevens-Johnson syndrome and toxic epidermal necrolysis (Kinoshita et al, Sci Transl Med, 2021). In the Cutaneous Leukocyte Biology Section at NIAMS, we further explore fundamental mechanisms that underlie tissue-immune and host-microbe interactions during homeostasis and inflammation. These processes involve innate and adaptive immune cells, and we hypothesize that altered crosstalk between epithelial or stromal compartments is involved in the pathophysiology of skin diseases. We employ clinically relevant models, including mouse models for AD and alopecia, to expand our fundamental knowledge of tissue-immune crosstalk and to build a foundation that will promote better understanding of skin immunity and inflammatory skin diseases.