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性和自身炎症性疾病的分子生物标志物。 我们还与Peter Grayson和Marcela Ferrada博士合作，他们正在研究复发性多囊炎（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，鼻和外周关节以及气管支气管树等组织中的反复炎症发作。炎症状态最终导致组织损伤和功能丧失。此外，这种疾病经常与风湿性关节炎有关。为了更好地了解RP患者发生的免疫学改变，我们对参加自然史研究的患者进行了广泛的免疫表型分析。 我们与Sarfaraz Hasni和Mariana Kaplan博士合作，参与了一项评估过氧化物酶体增殖物激活受体-γ激动剂吡格列酮（PGZ）在狼疮患者中的作用的研究。轻度至重度疾病活动的患者随机接受PGZ治疗，随后接受安慰剂治疗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进行）。目前在风湿病年鉴上发表的研究表明，PGZ耐受性良好，并在SLE患者中诱导血管僵硬度和心脏代谢参数的显着改善。结果表明，PGZ应进一步探索作为SLE心血管疾病风险的调节剂。 在与Dan Kastner博士和Christina Kozycki博士的合作研究中，我们参与了由ALPK 1基因中的显性功能获得性突变引起的自身炎症性疾病的表征，该突变导致了名为ROSAH的综合征（视网膜营养不良，视神经水肿，脾肿大，无汗症和头痛）。几乎所有患者（显著表型变异）均表现出至少一种与炎症一致的特征，包括反复发热、头痛伴脑膜增强和过早基底节/脑干矿化。TIS参与了信号研究，该研究表明患者表现出增加的STAT 1磷酸化，导致干扰素基因表达增强。抗TNF和抗IL-1治疗可抑制全身炎症并改善生活质量;抗IL-6（托珠单抗）是唯一可改善眼内炎症的抗细胞因子治疗。一份报告这些结果的手稿刚刚发表在《风湿病年鉴》上。 在与意大利罗马Sapienza大学的Giuseppe Scientin博士的合作中，我们发现转录因子STAT 4在小鼠肠道炎症期间调节ILC 1和NK细胞的效应分化方面发挥了意想不到的不同作用。描述这些结果的手稿目前正在审查中。 另一个富有成效的合作已经建立了与教授劳拉Santambrogio在威尔康奈尔医学院。在一项合作研究中，我们证明代谢损伤促进了来自蛋白质二硫键异构酶A家族成员3（PDIA 3）的巢状T和B细胞表位在MHC II类上的呈递，其参与免疫原性细胞死亡，最终支持致病性自身反应性。PDIA 3特异性T细胞或PDIA 3特异性抗体的被动转移加剧了肝细胞死亡，如通过增加的肝脏特异性转氨酶所确定的，在高脂肪小鼠而不是对照小鼠的血清中检测到。在慢性炎症性肝病患者中也观察到对PDIA 3的体液应答增加，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引起肝损伤，肝损伤通过提供PDIA 3 T细胞和B细胞表位促进致病性免疫自身反应性。一份描述这些结果的手稿目前正在《科学免疫学》杂志上出版。 TIS也一直在研究治疗自身免疫性疾病的新方法。我们目前正在与O 'Shea和Kaplan团队合作，评估托法替尼和第二代JAK选择性抑制剂对T细胞和先天性淋巴细胞的影响。由于小鼠JAK/STAT基因敲除研究的主要限制是先天淋巴细胞（ILC）群体（包括自然杀伤细胞）的完全丧失，因此用JAK抑制剂对这种信号级联的药理学改变是研究细胞因子作用的一种有吸引力的替代策略ILC生物学中的信号传导。我们一直在研究pan和JAK选择性抑制剂对不变的自然杀伤T细胞的发育和功能的影响，这些研究目前仍在进行中。

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.