Jarid2 regulates the function of follicular helper T cells and immune responses

免疫系统是人和动物体内帮助抵御入侵病原体和清除体内肿瘤细胞的防御系统，高等动物的免疫系统包括天然免疫系统和适应性免疫系统。适应性免疫可分为T细胞介导的细胞免疫应答和B细胞介导的体液免疫应答。生发中心（GerminalCenter，GC）反应是体液免疫应答中最重要的生理过程，活化的B细胞只有通过生发中心选择，才能分化成为分泌高亲和力抗体的浆细胞及长期记忆性B细胞，而生发选择过程是受到滤泡辅助性T细胞（FollicularhelperTcell，TFH）的控制。因此TFH细胞分化及功能的机制研究为体液免疫应答和免疫记忆形成提供重要的理论基础。 越来越多的研究表明，表观遗传修饰在各种免疫细胞的分化...The immune system protects host from invading pathogens and clears tumor cells. The vertebrate immune system is comprised of innate and adaptive immune system. Adaptive immunity can be classified into T cell-mediated cellular immunity and B cell-mediated humoral immunity. The germinal center reaction is the most important physiological process in humoral immunity. Only through germinal center sele...学位：理学硕士院系专业：生命科学学院_生物化学与分子生物学学号：2162014115254

Review of CD4+ T cell subsets in asthma phenotypes: molecular mechanisms and biologic treatment options

Asthma is a chronic inflammatory airway disease, mainly driven by different CD4+ T cell subsets [helper T cell (Th cell)]. CD4+ T cell subsets are a type of important immune cells, capable of secreting various cytokines, and regulating the immune response of the body to various antigens. According to the difference of secreted cytokines, CD4+ T cell subsets can be divided into subgroups such as Th1, Th2, Th17, follicular helper T cell (Tfh) and regulatory T cell (Treg), which play different roles in the occurrence and development of asthma. Biologic therapy is a new treatment method that targets specific molecules and pathways, and has provided more options for asthma patients. Biologics is a type of drugs prepared by biotechnology, which can specifically recognize and neutralize target molecules, thereby interfering with related signaling pathways. This article reviews the roles and molecular mechanisms of various CD4+ T cell subsets in asthma phenotypes, summarizes the immunopathological characteristics of eosinophilic asthma, neutrophilic asthma and mixed asthma, the clinical efficacy and safety of biologics targeting Th2, Th1, Th17, Tfh and Treg cell-related factors, and the selection and development direction of corresponding biologics. This article also discusses the role of impaired Treg cells and abnormal dendritic cells (DC cells) in the pathogenesis of asthma, as well as the potential of immunotherapy using these cells. This article aims to provide reference for the personalized selection and new drug development of biologic therapy for asthma

Leflunomide prolongs the survival time of xenograft in a rat-to-mouse heterotopic heart transplantation model

摘要 目的：异种移植是解决器官移植供体短缺问题的有效手段。在解决了超急性 排斥反应之后，目前急性血管性排斥反应（AVR）及细胞性排斥反应（CMR）的发生成为了异种移植应用于临床的最大免疫学障碍。因此，建立具有典型AVR及CMR表现的小动物异种移植模型，并在该模型基础上研究新型免疫抑制剂来氟米特（LEF）的作用机制十分重要。本实验的主要目的如下：1.首次运用套管法建立小鼠为受体的异种心脏移植模型2.比较以BALB/c及以C57BL/6小鼠为受体的两种模型的典型免疫排斥机制3.探究来氟米特对以上两种模型中典型免疫排斥的影响及作用机制。 方法：首先运用套管法建立Lewis大鼠——小鼠为...Abstract Backgrounds：Xenotransplantation is an effective way to circumvent donor shortage in clinical transplantation. However, there come AVR and CMR to be the greatest barrier for clinical use of xenotransplantation. Moreover, the immunosuppressant regime used clinically is inadequate to help surpass the immunological barrier. Our aim is to establish a rat-to-mouse heterotopic heart transpl...学位：医学硕士院系专业：医学院_外科学学号：2452014115353

Development of interleukin-21 as a therapeutic drug

白细胞介素21; (IL-21)是一种具有多效免疫调节活性的细胞因子,参与先天性免疫反应和适应性免疫反应的多个环节,与自身免疫性疾病、肿瘤、病毒感染等疾病的发病过; 程相关。研究表明,在动物模型中通过增强或抑制IL-21的生物学功能对多种疾病具有治疗意义,多项以重组人IL-21为受试药物的临床试验已经开展。本; 文就IL-21的生物学功能,以及IL-21作为生化药物在临床治疗中的研究进展和应用前景进行阐述。Interleukin-21 (IL-21) is a cytokine with broad pleiotropic actions that; plays an important role in both innate and adaptive immune responses.; IL-21 participates in pathogenesis processes of many diseases, such as; autoimmune disease, tumor, virus infections and so on. Enhancing or; inhibiting the activity of IL-21 has shown to have certain therapeutic; effects in animal models of a wide range of diseases. Various clinical; trials using recombinant human IL-21 as drug candidates are on-going; towards the development of IL-21 as a therapeutic drug. This article; reviewed the biological function and research progress and application; prospect of IL-21 as a biochemical drug in clinical treatment

Combining Monoclonal Antibodies with 1,25(OH)2D3 Treatment to Prolong the Survival Time of Cardiac Allograft in Accelerated Rejection Models of Mice

目的：供者反应性记忆性T细胞（memoryTcells，Tm）介导加速性排斥反应，是诱导移植物耐受的主要障碍。本研究利用1，25(OH)2D3和封闭性单克隆抗体anti-CD154/anti-LFA-1对Tm介导的加速性排斥反应进行治疗，比较不同用药组合对移植物的保护作用并探索该方案的作用机制。 方法：首先通过同种异体皮肤移植得到预致敏小鼠，过继转移同种反应性记忆性T细胞，构建小鼠同种异体心脏移植模型后再联合1，25(OH)2D3和anti-CD154/anti-LFA-1进行治疗，观察移植物生存期的改变，并从移植物和模型鼠整体两个角度探讨其可能的作用机制；之后以同种异体皮肤移植预致敏小鼠为...Aims: Donor-reactive memory T cells mediated accelerated rejection is known as a barrier to the survival of transplanted organs. In this study, we investigated the combination of 1,25(OH)2D3 and monoclonal antibodies anti-CD154/anti-LFA-1 to induce long-lived heart allograft acceptance in memory T cells-based adoptive mice model. The protective effects on grafts by different treatments were evalua...学位：理学博士院系专业：医学院基础医学系_生理学学号：2452009015372

N-of-1 Trial of Jianpiqingrehuashi Granular Decoction with Mesalamine in the Treatment of Ulcerative Colitis in Remission with Spleen Deficiency-induced Damp-heat Syndrome

BackgroundThe patient with ulcerative colitis (UC) in remission has a long course of disease, and needs a long-term maintenance treatment. Traditional Chinese medicine (TCM) has proved to be partially effective in treating UC in remission, but the efficacy of Jianpiqingrehuashi granular decoction (JPQRHSGD) against UC in remission is not yet clear.ObjectiveTo evaluate the efficacy and safety of JPQRHSGD with mesalamine versus mesalamine in the treatment of UC in remission using an N-of-1 trial.MethodsAn N-of-1 trial was conducted between June 2020 and March 2021. Participant was an outpatient with UC in remission with spleen deficiency-induced dampness-heat syndrome who was selected from the First Affiliated Hospital, Guangzhou University of Chinese Medicine. The patient received a treatment program, namely four rounds of treatment without washout consisting of eight cycles〔each round includes two cycles, that was, one-month intervention treatment (use of JPQRHSGD with mesalamine) alternating with one-month control treatment (taking mesalamine only) 〕. Efficaciesof two types of treatment were evaluated by of the TCM Syndrome Score (TCMSS) , Bristol Stool Form Scale (BSFS) , visual analogue scale (VAS) for abdominal pain and diarrhea, and Short Health Scale (SHS) . Safety was also compared between the two treatments.ResultsThe total TCMSS in intervention period was significantly lower than that in control period (P<0.05) . In particular, the symptoms of diarrhea, abdominal distension and fatigue of the limbs were improved more obviouslyin intervention period (P<0.05) . The improvements of total BSFS score, and diarrhea VAS score in intervention period were better than those in control period (P<0.05) . However, abdominal pain VAS score in intervention period was worse than that in control period (P<0.05) . The improvement of SHS score in intervention period was better than those in control period (P<0.05) . No treatment-related adverse events were reported.ConclusionThe JPQRHSGD with mesalamine improved the clinical symptoms of the UC patient in remission with spleen deficiency-induced dampness-heat syndrome, with relatively high safety

Immunological Mechanism of Faecal Bacterial Transplantation in the Intervention of Ulcerative Colitis

BackgroundUlcerative colitis is listed as one of the modern refractory diseases by the World Health Organization. At present, Western medicine still has many shortcomings in its treatment, and studies have shown that fecal bacterial transplantation (FMT) has a certain effect on it, but the mechanism is not clear.ObjectiveFMT was used to treat mouse ulcerative colitis model to verify the efficacy and possible mechanism of FMT.MethodsFrom December 2019 to April 2020, 60 mice were divided into normal control group (Control group) , ulcerative colitis model group (Model group) , ulcerative colitis model + fecal bacteria transplantation treatment group (Model+FMT group) and ulcerative colitis model + 5-aminosalicylic acid (5-ASA) treatment group (Model+5-ASA group) by random number table method, each of 15 mice. Control group did not make any intervention; Model group prepared mouse ulcerative colitis model; after successful modeling, the Model+FMT group was given 0.2 ml of fecal bacteria solution per enema; after successful modeling, the Model+5-ASA group was given 0.019 5 g/ml 5-ASA enema. The ultrastructural changes of intestinal tissue through transmission electron microscope, flow cytometric detection of blood helper T cell (Th) -17, Th-1, Th-2, Treg cell content changes, and enzyme-linked immunosorbent assay (ELISA) were observed to detect serum interferon γ (IFN-γ) , interleukin (IL) -2, IL-17, IL-4, IL-10, transforming growth factor β (TGF-β) level changes.ResultsIntestinal tissue transmission electron microscopy ultrastructure showed that the Model group was successfully modeled; the microvilli in the Model+FMT group and Model+5-ASA group were denser, with normal morphology, more goblet cells, slight swelling of mitochondria, and insignificant rough endoplasmic reticulum lesions. The Th17 cell content of the Model+FMT group was higher than that of the Control group and lower than that of the Model group; the Th17 cell content of the Model+5-ASA group was lower than that of Control group, Model group and Model+FMT group. The Th1 cell content of Model+FMT group and Model+5-ASA group were lower than those of Control group and Model group, respectively; Th2 cell content of Model+FMT group was lower than that of Control group and higher than that of Model group, and Th2 cell content of Model+5-ASA group was lower than that of Control group and higher than that of Model group and Model +FMT group. Treg cell content in Model+FMT group and Model+5-ASA group were lower than that of Control group and higher than that of Model group (P<0.05) . IFN-γ cell content in Model+5-ASA group was lower than that of Model group. IL-2 cell content in Model+FMT group and Model+5-ASA group was lower than that of Model group; the IL-17 cell content of Model+FMT group and Model+5-ASA group were lower than those of Control group and Model group, respectively. The IL-17 cell content of Model+5-ASA group was lower than that of Model+FMT group; the IL-4 cell content of Model+FMT group was lower than that of Control group and higher than that of Model group. The IL-4 cell content in the Model+5-ASA group was higher than that in the Model group; the IL-10 cell content in the Model+FMT group was higher than that in the Control group and the Model group, and the IL-10 cell content in the Model+5-ASA group was higher than that in the Model group; the content of TGF-β cells in the Model+FMT group and Model+5-ASA group were lower than those in the Control group and higher than those in the Model group (P<0.05) .ConclusionFMT can improve the symptoms of ulcerative colitis in mice. It is speculated that it may be achieved by adjusting the balance of Th1/Th2 cells and the ratio of Th17/Treg cells to achieve the purpose of treatment

细胞外囊泡参与调控外源因素诱导的肝毒性损伤的研究进展

肝脏是机体代谢外源因素的主要功能性器官，其具有独特的血窦结构和丰富的细胞组成，而细胞的功能改变是造成肝毒性损伤的主要原因。细胞外囊泡（extracellular vesicles, EVs）是由脂质双分子层所形成的纳米级球形囊泡，源自于各细胞亚群，用于负载特定的内容物，并可作为载体介导相邻或远处细胞间的信息交流。该文综述了外源因素暴露下EVs介导肝毒性损伤进程中各细胞的信息传递，并探讨靶向干预EVs的具体途径，为预防和控制肝毒性损伤提供线索以及潜在的应用价值。国家重点研发计划(2017YFA0205201)国家自然科学基金(81422023,51273165,U1705281,U1505221)教育部新世纪优秀人才支持计划(ncet-13-0502

外源物诱导肝脏局部先天免疫反应的线粒体调控机制

肝脏作为体内代谢外源因素暴露最重要的器官之一,具有独特的血窦结构和丰富的免疫细胞亚群,构成了肝脏局部免疫微环境。其中,肝脏的先天免疫细胞既可通过清除病原体、抗原提呈作用参与宿主防御,还通过与肝实质细胞的相互作用参与外源因素介导的急性免疫反应、肝细胞毒性转归、慢性肝损伤以及肝致癌作用。线粒体作为细胞应激的关键靶细胞器,是整合肝脏局部免疫信号的分子互作平台,既通过线粒体质量控制精细调控细胞内的分子事件,也通过线粒体损伤相关分子模式介导不同细胞间通讯和交互作用调控免疫微环境的稳态。本文通过对参与肝脏局部免疫反应的先天免疫细胞亚群组成及其介导免疫反应级联的综述,阐述不同外源物诱导肝脏局部免疫的线粒体相关调控机制,为探讨经由肝脏局部免疫反应生物标志筛查和靶向干预以预防和控制肝脏损伤提供线索

基于聚乳酸微球佐剂的乙肝疫苗研究

慢性乙肝的治愈之路充满挑战。与传统的抗病毒疗法相比，疫苗疗法是慢性乙肝预防和治疗的重要潜力途径。但是如何使疫苗获得有效的细胞免疫效果满足治疗性需求，具有更好的保护效果，是当前面临的重要挑战。本课题提出生物相容性好的聚乳酸（PLA）微球合理装载抗原和免疫刺激剂，实现按需递送的功能，作为治疗性乙肝疫苗，同时以PLA微球为佐剂探索免疫针次减少的疫苗制剂。课题具体研究内容如下：1）针对免疫刺激剂5,6-二甲基黄酮-4-乙酸（DMXAA）在特定细胞位置发挥作用的需求，成功制备了按需释放抗原的DP微球及抗原/刺激剂的DP-D微球，并验证了其在细胞水平的效果。通过向PLA微球中引入阳离子脂双十八烷基二甲基溴化铵（DDAB），微球可以温和高效携载乙肝表面抗原（HBsAg，吸附率85%以上）；并且借助质子海绵效应，促进溶酶体逃逸，使DMXAA释放到胞质中，与其受体结合，激活下游通路。在分子水平上，DP-D与抗原组相比，其STING通路的IRF-7和IFN-&beta;表达分别提高了12和8.2倍。同时微球携载HBsAg后易于被树突状细胞（DCs）摄取及引起溶酶体逃逸，提高DCs表面共刺激分子及MHC分子的表达，促进细胞因子分泌，在高效促进DCs活化的同时诱导后续免疫应答。2）评价了PLA微球佐剂在健康鼠上的体液和细胞免疫效果，探索了其应用于疫苗时的作用机制。微球疫苗制剂诱导了注射部位IL-1&beta;和CXCL-10等炎症因子和趋化因子的表达，同时招募DCs及巨噬等免疫细胞到注射部位，经APCs转运至淋巴结后，促进了淋巴结内DCs、B及滤泡辅助T细胞（Tfh细胞）的活化。在免疫健康鼠后，诱导产生了高水平的HBsAg特异性抗体IgG，提高IgG2a/IgG1比例、Th1细胞数量及IFN-&gamma;等细胞因子的分泌，实现了体液和细胞免疫的双重提升。3）成功构建了慢性乙肝（CHB）模型鼠，并基于该评价模型开展了微球疫苗制剂的免疫及治疗评价。采用尾静脉注射重组乙肝腺病毒的方式构建CHB模型鼠。结果表明，DP、DP-D组诱导产生高水平的HBsAg特异性抗体IgG，可促进脾细胞增殖活化与Th1型细胞因子的分泌，提升了HBsAg特异性CTL反应，产生免疫记忆起到保护效果。两组模型鼠血清中HBsAg转阴率均达到50%，显著降低了肝脏中HBcAg的表达，取得了良好的治疗效果。4）验证了以PLA微球为佐剂，用两针代替三针铝佐剂疫苗的长效免疫保护效果。微球疫苗制剂两针免疫后产生与铝佐剂相当的抗体水平，且抗体半年内维持较高水平，在HBsAg再次入侵时可快速产生抗体。优化了该疫苗制剂的冻干保护剂种类及相应浓度，考察了冻干制剂在室温下的稳定性。总体结果明确了微球疫苗佐剂具有减少免疫针次及可冻干的特性，具备临床转化的潜力。综上，本课题设计的PLA微球疫苗制剂应用于乙肝疫苗有较大潜力;Treatment of chronic hepatitis B (CHB) is full of challenge. Compared with traditional antiviral therapy, vaccine therapy is an important potential approach for the prevention and treatment of CHB. However, how to prime effective cellular immune response to meet the therapeutic requirements and endow a better protective effect is a great challenge. This dissertation proposes that biocompatible polylactic acid (PLA) microparticles (MPs) which rationally loaded with antigen and immunopotentiator according to the delivery demand to act as therapeutic hepatitis B vaccine. Meanwhile, PLA MPs were used as adjuvants to explore vaccines with reduced immunization dose. In detail, this thesis mainly includes the following issues: 1) Aim at the immunopotentiator need to function at specific cell locations, DP and DP-D MPs were successfully fabricated according to the release demand of antigen and immunopotentiator, and evaluated the immune response at cellular level. The introduction of cationic lipid didodecyldimethylammonium bromide (DDAB) into PLA MPs could assist them adsorb antigen gently and efficiently (adsorption rates were all above 85%); meanwhile DMXAA could release into the cytoplasm through the proton sponge effect, then interact with its receptor, activate downstream pathway. At the molecular level, the expression of IRF-7 and IFN-&beta; in the STING pathway of DP-D increased by 12 and 8.2 times respectively compared with the antigen group. The MPs vaccines could facilitate cell uptake and lysosome escape, increase the expression of costimulatory and MHC molecules of dendritic cells (DCs), promote cytokines secretion, thus activating DCs and prime subsequent immune responses efficiently. 2) The humoral and cellular immune responses of PLA MPs vaccines on healthy mice were evaluated and the immunization mechanism was explored as vaccine adjuvant. MPs vaccines can induce the expression of inflammatory cytokines and chemokines such as IL-1&beta; and CXCL-10 at the injection site, and recruit immune cells such as DCs and macrophages to the injection site. After being transported to lymph nodes by APCs, they can promote DCs, B and Tfh cells activation in lymph nodes. In healthy mice immunization experiment, high levels of HBsAg-specific IgG and IgG2a / IgG1 ratio were induced, the number of Th1 splenocytes, and the secretion of IFN-&gamma; together with other cytokines were also elevated, indicating the MPs induce both humoral and cellular immune responses. 3) The chronic hepatitis B (CHB) model mice were successfully constructed, and the immune and therapeutic effect of the MPs vaccines were investigated. CHB model mice were constructed by tail vein injection of recombinant AAV/1.3HBV virus. The results indicated that DP and DP-D induced high-level HBsAg-specific IgG, promoted the splenocytes proliferation and activation as well as the secretion of Th1 cytokines. The HBsAg-specific CTL responses were improved and memory immune responses for protection were maintained. The serum HBsAg seroconversion rate of the two MPs vaccines both achieved 50% and the expression of HBcAg in the liver significantly reduced, indicating a favorable therapeutic effect. 4) The long-term immunoprotection effect of two dose PLA MPs vaccines instead of three dose aluminum vaccines were verified. The MPs vaccines produced comparable antibodies compared to aluminum adjuvant after two dose immunization schedule, and the antibodies maintained at a high level for half a year. When rechallenging with HBsAg, the mice can quickly produce antibodies. The type and corresponding concentrations of lyoprotectants were optimized, and the stability of lyophilized vaccines at room temperature was investigated. The results clarified that the MPs vaccine adjuvant has the capacity of reducing the immunization dose and can be lyophilized, thus possessing the potential for clinical transformation. In summary, the PLA MPs vaccines designed in this dissertation possess great potential for application in hepatitis B vaccines.&nbsp;</p