Translational Immunology research: a support for clinical immunological research

涉及TIS的一个主要项目是定义自身免疫和自体炎症性疾病的分子生物标记物。 我们还与Peter Grayson博士和Marcela Ferrada博士合作，他们正在研究导致复发性多软骨炎(RP)的分子机制，RP是一种免疫介导的炎症性全身性疾病，其特征是耳朵、鼻子、外周关节以及气管支气管树等组织反复发作炎症。炎症状态最终会导致组织损伤和功能丧失。此外，这种疾病经常与类风湿性关节炎有关。为了更好地了解RP患者中发生的免疫学变化，我们对参加自然病史研究的患者进行了广泛的免疫表型分析。 在Sarfaraz Hasni博士和Mariana Kaplan博士的合作下，我们参与了一项研究，该研究评估了狼疮患者中的过氧化体增殖物激活受体-伽玛激动剂吡格列酮(PGZ)。有轻度到重度疾病活动的患者被随机分成一系列的PGZ，然后是安慰剂，为期3个月，反之亦然，采用双盲交叉设计，洗脱期为2个月。测量内皮功能和动脉炎症的参数以及中性粒细胞失调、代谢紊乱和基因表达研究(由TIS执行)。目前发表在《风湿病年鉴》上的这项研究表明，PGZ耐受性良好，并导致SLE患者血管僵硬和心脏代谢参数的显著改善。研究结果提示，PGZ作为系统性红斑狼疮心血管疾病风险的调节因子值得进一步探讨。 在与Dan Kastner博士和Christina Kozycki博士的合作研究中，我们参与了由ALPK1基因的显性功能获得突变引起的自体炎症性疾病的特征描述，该突变导致了一种名为ROSAH的综合征(视网膜营养不良、视神经水肿、脾肿大、无汗和头痛)。几乎所有的患者(显著的表型变异)至少表现出一个与炎症相一致的特征，包括反复发烧、头痛伴脑膜强化和基底节/脑干矿化过早。TIS参与了信号研究，研究表明患者表现出STAT1磷酸化增加，导致干扰素基因表达增强。抗肿瘤坏死因子和抗IL-1治疗抑制全身炎症，改善生活质量；抗IL-6(托西珠单抗)是唯一能改善眼内炎症的抗细胞因子治疗。一篇报道这些结果的手稿刚刚发表在《风湿病年鉴》上。 与意大利罗马萨皮恩扎大学的朱塞佩·肖姆博士合作，我们发现转录因子STAT4在小鼠肠道炎症过程中，在调节ILC1和NK细胞的效应器分化方面发挥了出人意料的不同作用。描述这些结果的手稿目前正在审查中。 与威尔·康奈尔医学院的劳拉·桑坦布罗吉奥教授建立了另一项富有成效的合作。在一项合作研究中，我们证明了代谢损伤促进了来自蛋白二硫键异构酶家族成员3(PDIA3)的嵌套T和B细胞表位的呈现，这参与了免疫原性细胞死亡)，最终支持了致病的自身反应。被动转移PDIA3特异性T细胞或PDIA3特异性抗体会加剧肝细胞死亡，这是通过在高脂小鼠而不是对照组小鼠的血清中检测到的肝脏特异性转氨酶的增加来确定的。在慢性炎症性肝病患者中也观察到对PDIA3的体液反应增加，包括自身免疫性肝炎、原发性胆管炎和2型糖尿病。研究表明，高脂高果糖饮食引起的代谢损伤导致肝脏损伤，通过提供PDIA3 T细胞和B细胞表位促进病原性免疫自身反应。目前正在出版的《科学免疫学》杂志上有一篇描述这些结果的手稿。 TIS还一直在研究治疗自身免疫性疾病的新方法。在与O‘Shea和Kaplan小组的合作下，我们目前正在评估tofacitinib和第二代JAK选择性抑制剂对T细胞和固有淋巴样细胞的影响。由于JAK/STAT基因敲除研究的一个主要局限性是先天淋巴样细胞(ILC)群体(包括自然杀伤细胞)的完全丧失，因此将这种信号级联与JAK抑制剂进行药理交替是研究细胞因子信号在ILC生物学中作用的一个有吸引力的替代策略。我们一直在研究PAN和JAK选择性抑制剂对不变自然杀伤T细胞发育和功能的影响，目前研究仍在进行中。

A major project that involves the TIS is the definition of molecular biomarkers for autoimmune and autoinflammatory diseases. We are also collaborating with Drs. Peter Grayson and Marcela Ferrada who are investigating the molecular mechanisms driving Relapsing polychondritis (RP), an immune-mediated, inflammatory, systemic disorder characterized by recurrent episodes of inflammation in tissues such as the ear, nose and peripheral joints as well as the tracheobronchial tree. The inflammatory status eventually results in tissue damage and functional loss. Moreover, the disease is frequently associated with Rheumatoid Arthritis. To better understand the immunological alterations occurring in RP patients, we have extensively immunophenotyped the patients that enrolled in their natural history study. In collaboration with Drs. Sarfaraz Hasni and Mariana Kaplan we have been involved in a study that assessed the peroxisome proliferator activated receptor-gamma agonist pioglitazone (PGZ) in Lupus patients. Patients with mild to severe disease activity were randomized to a sequence of PGZ followed by placebo for 3 months, or vice versa, in a double-blind, cross-over design with a 2-month wash-out period. Parameters of endothelial function and arterial inflammation were measured as well as neutrophil dysregulation, metabolic disturbances, and gene expression studies (performed by the TIS). The study that is currently in press in Annals of Rheumatic Disease showed that PGZ was well tolerated and induced significant improvement in vascular stiffness and cardiometabolic parameters in SLE. The results suggest that PGZ should be further explored as a modulator of cardiovascular disease risk in SLE. In a collaborative study with Dr. Dan Kastner and Dr. Christina Kozycki we participated in the characterization of an autoinflammatory disease caused by dominant gain-of-function mutations in the ALPK1 gene which resulted in a syndrome named ROSAH (retinal dystrophy, optic nerve oedema, splenomegaly, anhidrosis and headache). Nearly all patients (significant phenotypic variation) exhibited at least one feature consistent with inflammation including recurrent fever, headaches with meningeal enhancement and premature basal ganglia/brainstem mineralization. The TIS was involved in signaling studies which showed that patients exhibited increased STAT1 phosphorylation leading to augmented interferon gene expression. Anti-TNF and anti-IL-1 therapies suppressed systemic inflammation and improved quality of life; anti-IL-6 (tocilizumab) was the only anti-cytokine therapy that improved intraocular inflammation. A manuscript reporting these results has just been published in Annals of Rheumatic Diseases. In collaboration with Dr. Giuseppe Sciume at Sapienza University in Rome, Italy, we have shown that the transcription factor STAT4 played an unexpectedly divergent role in regulating effector differentiation of ILC1 and NK cells during intestinal inflammation in mice. A manuscript describing these results is currently under review. Another fruitful collaboration has been established with Professor Laura Santambrogio at Weill Cornell School of Medicine. In one collaborative study we demonstrated that metabolic insults promoted the presentation of nested T and B cell epitopes from protein disulfide isomerase family A member 3 (PDIA3), which is involved in immunogenic cell death) on MHC Class II, ultimately supporting pathogenic autoreactivity. Passive transfer of PDIA3-specific T cells or PDIA3-specific antibodies exacerbated hepatocytes cell death, as determined by increased liver-specific transaminases, detected in the sera of high fat but not control mice. Increased humoral responses to PDIA3 were also observed in patients with chronic inflammatory liver conditions including autoimmune hepatitis, primary biliary cholangitis, and type 2 diabetes. The study showed that metabolic insults caused by high-fat-high-fructose diet elicited liver damage, which promotes pathogenic immune autoreactivity by supplying PDIA3 T cell and B cell epitopes. A manuscript describing these results in currently in press in the journal Science Immunology. The TIS has also been investigating novel approaches for the treatment of autoimmune diseases. In collaboration with the O'Shea and Kaplan groups we are currently evaluating the effects of tofacitinib and second-generation JAK-selective inhibitors on T cells and innate lymphoid cells. Since a major limitation of JAK/STAT gene knockout studies in mice is the complete loss of Innate Lymphoid Cell (ILC) populations (including natural killer cells), pharmacological alternation of this signaling cascade with JAK inhibitors is an attractive alternative strategy to study the role of cytokine signaling in ILC biology. We have been investigating the effects of pan and JAK-selective inhibitors on the development and functions of invariant natural killer T cells and the studies are still currently ongoing.