Mast cells play a central role in allergic reactions, acting as key effector cells that initiate and amplify the inflammatory response. In this study, we demonstrate that phosphatase of regenerating liver 2 (PRL2) functions as a negative regulator of FcεRI-mediated mast cell activation. In PRL2-deficient myeloid cells, PRL2 conditional knockout mice developed more severe passive systemic anaphylaxis (PSA). Although PRL2 deficiency does not impact mast cell development, in the absence of PRL2 FcεRI-mediated mast cell activation is enhanced. In the presence of IgE the expression of mast cell PRL2 is downregulated, leading to modulation of the cellular response. In PRL2-deficient mast cells, the PI3K signaling pathway is upregulated, resulting in increased calcium influx. This, in turn, enhances mast cell degranulation and the production of inflammatory mediators. Moreover, hydroxychloroquine (an inhibitor of PRL2 degradation) reduces the severity of PSA in wild-type mice. Our findings suggest that PRL2 acts as a negative regulator of FcεRI-mediated mast cell activation. Therefore, therapeutic strategies aimed at enhancing PRL2 activity in mast cells may offer a promising approach for the treatment of allergic disorders.

Mast cells are phenotypically and functionally heterogeneous, and their state is possibly controlled by local microenvironment. Therefore, specific analyses are needed to understand whether mast cells function as powerful participants or dispensable bystanders in specific diseases. Here, we show that degranulation of mast cells in inflammatory synovial tissues of patients with rheumatoid arthritis (RA) is induced via MAS-related G protein-coupled receptor X2 (MRGPRX2), and the expression of MHC class II and costimulatory molecules on mast cells are upregulated. Collagen-induced arthritis mice treated with a combination of anti-IL-17A and cromolyn sodium, a mast cell membrane stabilizer, show significantly reduced clinical severity and decreased bone erosion. The findings of the present study suggest that synovial microenvironment-influenced mast cells contribute to disease progression and may provide a further mast cell-targeting therapy for RA.

Abstract Objectives The ‘Host Defense and Immunology’ course is compulsory for undergraduate students majoring in clinical medicine at Shanghai Jiao Tong University School of Medicine. In order to promote students’ self-directed and active learning, we innovated teaching activities around ‘seamless learning’ which is emphasising learner-centredness. Methods Guided by the concept of seamless learning, we integrated teaching content, constructed course resources, innovated course-teaching modes, and reformed assessment and evaluation. Then two survey questionnaires were provided to students to get feedback on the course achievement. One set of survey questionnaire with written descriptions was designed to collect feedback on the overall evaluation of the course. Another survey questionnaire was presented in the form of multiple-choice questions to get feedback on more specific questions. Results 294 questionnaires were distributed to eight-year clinical medicine students in years 2021–2022, and 234 were valid. Overall student feedback revealed that the students mastered the immunological basic theories, research techniques and have acquired integrated systematic thinking. The students also deepened their application ability, pursued clinical rigor and logic, and shaped their clinical thinking patterns. The survey questionnaire revealed that the comprehensive immunological mind map was highly beneficial for analysing clinical immunological problems, understanding immunological principles, grasping interrelationships between knowledge points, and fostering self-learning abilities. Case-based learning (CBL), micro-lecture videos, and learning about scientists’ and doctors’ stories also had positive effects on students’ active learning, understanding, logical thinking, innovation, and humanistic literacy, with over 79 % of students reporting benefits. Conclusions Based on seamless learning, students acquired integrated systematic thinking through the course, shaped clinical thinking patterns, developed communication and collaboration skills, and enhanced their ability to analyse and solve clinical problems. The exploration and practice of the ‘Host Defense and Immunology’ course provides a good example for the integration of seamless learning concepts into teaching.

Psoriatic arthritis (PsA) is a disease that transformed from psoriasis (PsO), and its underlying mechanisms are still not fully understood. Overactivation of the immune system is a key factor driving inflammatory diseases. Our goal is to define the unbalanced subsets of peripheral blood CD4 +T cells between PsO and PsA patients. Blood samples from 43 patients (23 PsA and 20 PsO) and 36 healthy donors (HD) were studied. Peripheral blood mononuclear cells (PBMC) were separated from blood and underwent fluorescent staining to assess CD4+T cell subsets by flow cytometry. We found that frequencies of various CD4+T cells including Th1, Th2, Th17, and Tfh were higher in the patients with PsO or PsA than those of healthy donors, indicating the general expansion of CD4+T cells in inflammatory conditions. More importantly, we observed the significant imbalance of Th1/Th2 between patients with PsO and PsA. Pearson correlation analysis showed that Th1/Th2 ratio was positively correlated with disease activity in psoriatic arthritis (DAPSA), Tfh/Tfr ratio was positively correlated with DAPSA score and visual analogue scale (VAS) score in PsA patients. Together, our results highlight the CD4+T cell changes in the transition from PsO to PsA, may contribute to early assessment and intervention.

Fibroblast-like synoviocytes (FLS) mediate many pathological processes in rheumatoid arthritis (RA), including pannus formation, bone erosion, and inflammation. RA FLS have unique aggressive phenotypes and exhibit several tumor cell-like characteristics, including hyperproliferation, excessive migration and invasion. Casein kinase 2 (CK2) is reportedly overexpressed in numerous tumor types, and targeted inhibition of CK2 has ther-apeutic benefits for tumors. However, the expression level of CK2 and its functions in RA FLS remain unclear. Herein, we aimed to elucidate whether CK2 is responsible for the aggressive phenotypes of RA FLS and whether targeted therapy can alleviate the severity of RA. We found that CK2 subunits were elevated in RA FLS compared with osteoarthritis FLS, and the activity of CK2 also markedly increased in RA FLS. Targeted inhibition of CK2 using CX-4945 suppressed RA FLS proliferation through cell cycle arrest. Cell migration and invasion were also inhibited by CX-4945 treatment. Moreover, CX-4945 reduced Interleukin-6 (IL-6), C-C motif chemokine ligand 2 (CCL2) and Matrix metalloproteinase-3 (MMP-3) secretion in RA FLS. Further proteomic investigation revealed that p53 signaling pathway significantly changes after CX-4945 treatment in RA FLS. The siRNA-mediated p53 knockdown partly abolished the anti-proliferation and reduced IL-6, MMP-3 secretion effects of CX-4945. Furthermore, CX-4945 administration alleviates arthritis severity in CIA mice. Collectively, our results demon-strated the abnormal elevation of CK2 and its positive association with abnormal phenotypes in RA FLS. Our novel findings suggest the possible therapeutic potential of CX-4945 for RA.

"Host Defense and Immunity" is an integrated course for medical undergraduates in Medical School of Shanghai JiaoTong University, which trains students "excellent medical innovative talents with soul" from aspects of knowledge, ability and quality. Host Defense and Immunity has won honor of the first batch of national first-class undergraduate offline courses. This paper introduces construction and practical experience of this course in ideological and political aspects, adds corresponding ideological and political objectives, condense value shaping point, organizes and compiles corresponding ideological and political cases and shoots a series of micro-videos, and practices combination of ideological and political courses and majors in teaching, discussion and usual assessment, so as to achieve educational purpose of "Building morality and cultivating people".

γ-Tocotrienol (GT3), a member of the vitamin E family, is well known for its medicinal value in clinical treatments. However, the role of GT3 in T helper 17 (Th17)/regulatory T cell (Treg) differentiation and function is not fully understood. Here, we demonstrated that GT3 suppressed Th17 differentiation in vitro by inhibiting signal transducer and activator of transcription 3 (STAT3) phosphorylation in the interleukin 6 (IL-6)/Janus kinase (JAK)/STAT3 signaling pathway. GT3 also inhibited HIF1A expression in Th17 metabolism. Additionally, we showed that GT3 treatment inhibited disease aggravation in an imiquimod (IMQ)-induced psoriasis-like mouse model by reducing the percentage of Th17 cells in the spleen in vivo. The findings of this study demonstrated the effects of GT3 on Th17 cells through the STAT3 signaling pathway.

Epigallocatechin-3 gallate (EGCG) is a polyphenolic component of tea and has potential curative effects in patients with autoimmune diseases. Multiple sclerosis (MS) is an autoimmune disease affecting the central nervous system (CNS). It remains unknown whether EGCG can regulate macrophage subtypes in MS. Here we evaluated the effects of EGCG in experimental autoimmune encephalomyelitis (EAE), MS mouse model. We found that EGCG treatment reduced EAE severity and macrophage inflammation in the CNS. Moreover, EAE severity was well correlated with the ratio of M1 to M2 macrophages, and EGCG treatment suppressed M1 macrophage-mediated inflammation in spleen. In vitro experiments showed that EGCG inhibited M1 macrophage polarization, but promoted M2 macrophage polarization. These effects were likely to be related to the inhibition of nuclear factor-κB signaling and glycolysis in macrophages by EGCG in macrophages. Overall, these findings provided important insights into the mechanisms through which EGCG may mediate MS.

Multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE) are neuroinflammatory autoimmune diseases characterized by the axonal loss, demyelination, and neurodegeneration of the central nervous system. Overactivation of CD4+ T cells, especially the migration of the Th1 and Th17 subsets into the central nervous system (CNS), leads to the secretion of inflammatory mediators and destruction of the contact between neurons and activated macrophages, which can then result in a series of neurocognitive and motor deficits. In this study, we intended to explore the role of miRNA-467b in regulating Th cell development in EAE. We found that the level of miRNA-467b was decreased and eukaryotic initiation factor 4 F (eIF4E) was increased in lymph nodes and the CNS at EAE peak. eIF4E was confirmed as the direct target of miRNA467b. Overexpression of miRNA-467b could suppress a percentage of CD4+ IL-17+ cells in EAE CD4 + T cells in vitro. In addition, we also identified miRNA-467b, which could suppress Th17 cell differentiation by targeting eIF4E in vitro. Furthermore, injecting miRNA-467b mimics into the caudal vein of EAE mice contributed to less inflammation in the peripheral lymphoid organs and CNS and alleviated disease severity. Taken together, our findings imply that miRNA-467b inhibits the differentiation and function of Th17 cells by targeting eIF4E, thereby alleviating EAE.

目的·探讨类风湿关节炎(rhumatoid arthritis,RA)患者B细胞亚群特点及表没食子儿茶素没食子酸酯(epigallocatechingallate,EGCG)对RA患者B细胞亚群的调节作用.方法·纳入年龄、性别匹配的RA患者和健康对照各29例,运用配对t检验分析2组外周血中B细胞亚群的差异;根据28关节疾病活动评分(disease activity score in 28 joints,DAS28)的值将RA患者分为活动组(2.6≤DAS28<5.1)和高度活动组(DAS28≥5.1),运用t检验分析2组外周血中B细胞亚群的差异.在0、10、100μmol/L EGCG和2.5μg/L葡萄球菌A蛋白共刺激条件下,体外培养RA患者外周血单个核细胞,24 h后应用实时聚合酶链反应检测B细胞活化因子受体(B-cell-activating factor receptor,BAFF-R)的表达水平,48 h后用流式细胞术检测B细胞亚群.结果 ·RA患者的总B细胞、未分化B细胞、记忆B细胞和浆母细胞占淋巴细胞比例和数量均高于健康对照(P<0.05),而CD19+IL-10+调节性B细胞(regulatory B cell,Breg)占淋巴细胞比例和数量与健康对照相比差异无统计学意义(P>0.05).10例活动组与19例高度活动组RA患者的总B细胞及各B细胞亚群(除CD19+IL-10+Breg外)占淋巴细胞比例和数量的差异无统计学意义(P>0.05),6例活动组与12例高度活动组RA患者CD19+IL-10+Breg占淋巴细胞比例和数量的差异也无统计学意义(P>0.05).RA患者外周血总B细胞所占比例与IgG型类风湿因子呈弱正相关(r=0.308).EGCG能显著提高CD19+IL-10+Breg占淋巴细胞比例(P<0.05),且100μmol/L EGCG能显著降低总B细胞中BAFF-R mRNA的表达水平(P=0.000);但其对未分化B细胞、记忆B细胞和浆母细胞占淋巴细胞比例无显著影响(P>0.05).结论 ·B细胞可能在RA发病中发挥辅助作用,RA患者CD19+IL-10+Breg数量反馈性增加,EGCG可促进Breg增殖,降低BAFF-R mRNA表达水平.

目的·探讨miR-322-5p靶向Akt3抑制Th17分化对干扰素 β(interferon-β,IFN-β)干预实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)的影响.方法·建立EAE小鼠模型,设IFN-β 干预组和PBS对照组.流式染色比较2组Th17的比例变化;RNA芯片检测2组小鼠miRNA的差异表达,筛选出miR-322-5p做进一步研究;软件预测miR-322-5p的靶基因为Akt3;IFN-β 干预后和过表达miR-322-5p后检测Akt3的表达水平;双荧光素酶报告实验验证miR-322-5p和Akt3的直接靶向关系;体外实验观察Akt3对Th17细胞分化的影响.结果 ·IFN-β 干预组的EAE小鼠Th17比例均显著降低,miR-322-5p的表达显著升高,而Akt3的表达明显降低;过表达miR-322-5p能显著抑制Akt3的表达,双荧光素酶报告实验显示Akt3是miR-322-5p的直接靶基因,且Akt3对Th17的体外分化有明显促进作用.结论 ·IFN-β 可通过影响miR-322-5p靶向Akt3,进而抑制Th17分化来缓解EAE的疾病进程.

微课程教学是一种新的教学形式,能够提高学生的学习兴趣和效率,培养学生自主学习的能力.免疫学的教学内容具有概念抽象、逻辑性强、知识增长快等特点.因此,采用微课程教学作为其传统教学方式的辅助是颇有成效的一种尝试.本文总结了免疫学微课程教学的特色;分析了国内现阶段免疫学微课程教学中的不足及可采取的对策;并针对如何进行优质的免疫学微课程教学提出了建议.免疫学中的微课程教学值得进一步的尝试、完善和推广.

为了比较阳离子脂质体Lipofectamine 3000 (Lipo3000)在不同细胞类型中的转染效率,课题组分别在贴壁细胞NIH3T3、悬浮细胞EL4和原代细胞CD4+T细胞中,利用Lipo3000将不同浓度的miRNA mimics转染至细胞内,通过流式细胞术检测的方法,比较不同细胞之间以及同一细胞、不同浓度的miRNAmimics组之间转染效率的差异.结果显示,在NIH3T3细胞中,miRNAmimics为100 nmol/L时,转染效率最高,可达(41.47±8.10)％;而在EL4和CD4+T细胞中,随着miRNA mimics浓度的增加,转染效率呈上升的趋势,最高分别可达(12.13±1.16)％和(3.80±0.60)％.可见,Li-po3000在贴壁细胞和悬浮细胞的转染效率明显高于原代细胞.

为解决免疫学教学中存在的一些问题,设计综合思维导图教学方式,试图将抽象原理可视化、众多概念结构化、分散知识点系统化,使分散在各章的概念与原理梳理成相互联系的知识模块并成为整体,调动学生的主动学习,促进对原理的理解和记忆,训练逻辑思维和创新性思维.初步运用后调查显示,对自主学习能力训练、各种概念和原理之间关系梳理及免疫学原理的整体理解等很有帮助,对名词的理解和记忆等则效果略差,对批判性思维的训练则无效果.

机体防御与免疫课程属于基础医学的横向整合课程之一,对课程内容、教学形式和考核方式均进行了改进.文章分析了国内外医学免疫学课程的实施现状,总结了该课程的实践成果.

Multiple sclerosis (MS) and the corresponding animal model, experimental autoimmune encephalomyelitis (EAE), are chronic neuroinflammatory autoimmune diseases. Increased activation of CD4+T cells, especially the Th1 and Th17 subsets, is thought to play a causal role in this disease. IFN-β is widely used in the treatment of MS and is found to decrease IL-17 and OPN production in MS patients and EAE mice. However, a definitive molecular mechanism has not yet been fully elucidated. In this study, we investigated the immunomodulatory effect of IFN-β on the EAE model. We observed disease progression and determined the percentage of Th1/Th17 cells in the peripheral immune organs, brain, and spinal cord of mice. Furthermore, the levels of related cytokines and transcription factors were measured in splenocytes, and the effects of IFN-β on Th17 differentiation were assessed in vitro. Compared to the control group, IFN-β treatment significantly reduced the incidence of EAE and the associated pathological damage. Th1 and Th17 cells in IFN-β-treated mice were significantly reduced, and the levels of cytokines, such as IFN-γ, IL-17, and OPN, were significantly decreased in splenocyte supernatants as well as the levels of corresponding transcription factors. IFN-β inhibited downstream inflammatory cytokines through the inhibition of PI3K/AKT/NF-κB axis and p38, JNK-MAPK, as well as the regulation of mTOR complexes. Moreover, IFN-β inhibited Th17 differentiation and neutralizing OPN antibodies offset the inhibitory effect of IFN-β on Th17 cells. Meanwhile, IFN-β influenced the acetylation of the Il17a and Opn gene promoters. The findings described herein provide novel evidence for the role of IFN-β in Th17 differentiation partly through the inhibition of OPN.

Rheumatoid arthritis (RA) is an autoimmune inflammatory disease characterized by bone loss. Degree of inflammation has been identified as an important initiator of skeletal damage in RA. Iguratimod (T-614) is an anti-inflammatory agent which has been reported to show the inhibitory effect of bone destruction in RA. However, the role of T-614 in osteoblast differentiation is still not clear. In this study, we intended to find the effect of T-614 on the osteogenesis process. We detected osteogenesis markers and transcription factors associated with osteoblastic lineage and bone formation in the culture of mesenchymal stem cells which differentiate osteoblast. The contents and activity of alkaline phosphatase, levels of collagen type I and bone gla protein, and calcium nodule formation were increased significantly after T-614 treated. Meanwhile, the mRNAs expressions of Osterix and Dlx5 were also found to be increased significantly by real-time PCR. The changes of levels of phosphorylation of p38 and NF-κB were also detected by Western blot. The results showed that T-614 promotes osteoblastic differentiation by increasing the expression of Osterix and Dlx5 and increasing the activation of P38. T-614 could advance the ectopic expression of NF-κB to suppress inflammation, which indirectly inhibits the damage of the osteoblasts.

为探究强直性脊柱炎(ankylosing spondylitis,AS)的发病机制和免疫细胞在病情发展中的功能和重要性,我们设计本实验以比较强直性脊柱炎患者与正常人外周血中免疫细胞格局的变化.实验以42例AS患者和42例健康者为研究对象,检测外周血T细胞以及相应的细胞因子和转录因子.实验采用流式细胞术检测T细胞亚群的比例、通过ELISA方法检测细胞因子并使用Q-PCR检测转录因子的表达.实验发现,与健康人相比,AS患者外周血T细胞中CD4+细胞占优势,并且effect T细胞与naive T细胞的比值升高;对CD4+T细胞亚群进一步的检测结果显示Th1与Tfh的比例显著增高,Treg 细胞的比例较正常人稍高.在对细胞因子的检测中,我们发现IFN-γ、TGF-β显著下降,但IL-21有上升趋势;对转录因子的检测显示,Bcl-6/Foxp3、RORγt/Foxp3和T-bet/Foxp3值均升高.上述结果提示强直性脊柱炎患者外周血中T细胞亚群的格局发生了变化,其相关免疫分子也呈现免疫活化状态.

We aimed to investigate the role of miRNA-467b in inflammatory T cell migration in EAE.Firstly,the expressions of CXCR3,CCR5,CCR2 and CCR6 in EAE splenocytes were measured by qRT-PCR and FACS.Secondly,synthetic miRNA-467b mimics or normal control(NC) were transfected into MOG3s-ss-specific T cells by liposome method,and the levels of CXCR3,CCR5,CCR2 and CCR6 were detected by FACS.Finally,to further explore the effect of miRNA-467b on the migration of MOG35-55-specific T cell,cell migration assay was detected by Boyden chemotaxis chamber.The results showed that:1.The expressions of CXCR3,CCR5,CCR2 and CCR6 in EAE splenocytes were higher than those in control mice(P＜0.05).2.miR-NA-467b overexpression could reduce the levels of chemokine receptors in Th1 cells(P＜0.05).3.Cell migration assay revealed that miRNA-467b can effectively reduce the migration of MOG3s-s5-specific T cells(P＜0.05).In conclusion,miRNA-467b could decrease the levels of chemokine receptors in Th1 cells and reduce the migration of inflammatory T cells,which indicates that miRNA-467b may play an important role in EAE disease process.

Rheumatoid arthritis (RA) is a systemic autoimmune disease that results in a chronic and inflammatory disorder. Dynamic balance of helper T cells (Th) 1 and 17 and regulatory T cells (Treg) is broken in RA. Since there is no cure for RA at present, it is necessary to find a truly effective and convenient treatment. Several studies have intended to induce ergotopic regulation to treat autoimmune diseases. This study was undertaken to find potential ergotope peptides and investigate their effects in treating the animal model of RA and their underlying regulatory mechanisms. First, we selected functional ergotope peptides from 25 overlapping peptides derived from the interleukin 2 receptor (IL-2R) α chain, and then used these peptides to treat collagen-induced arthritis (CIA). We showed ergotope peptides as immunomodulatory factors with great benefits at the clinical and pathologic levels. This effect was associated with inhibition of type II collagen (CII)-specific proliferation and autoantibody production as well as induction of antiergotypic immune response, downregulation of both Th1 and Th17 cells and their related components, and emergence of Treg cells that had suppressive action on autoreactive T cells. We also proved that cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) and IL-10 are two important mediators that are critical to Treg suppressive function. Inhibition of Th1 and Th17 in established CIA could be attributed to ergotope-induced Treg cells. Our findings reveal that ergotope peptides induce regulatory immune responses and restore immune tolerance, suggesting that treatment with ergotope peptides may be a novel approach to therapy for RA patients and has good application prospects, with cheap, effective, convenient, wide-spectrum features.