Rheumatoid arthritis (RA) is characterized by chronic synovitis, leading to joint cartilage and bone erosion. While current treatments aim to control inflammation, no specific drug prevents joint structure damage. This study set out to explore the efficacy of (5R)-5-hydroxytriptolide (LLDT-8), which targets osteoclasts known to be closely related to RA-induced erosion. We evaluated morphological changes of bone marrow cells and identified the induced osteoclasts by TRAP staining. LLDT-8’s inhibitory effect on the bone erosion activity of osteoclasts was assessed by resorption pit assay and the induction of apoptosis was evaluated by flow cytometry. Immunohistochemical staining was performed to detect the vascular endothelial growth factor (VEGF) expression in osteoclasts. LLDT-8 inhibited the transformation of bone marrow cells into osteoclast cells and induced the apoptosis of osteoclasts. Moreover, the number of osteoclasts-induced bone lacunae and the expression of VEGF expressions of osteoclasts were significantly reduced by LLDT-8. We demonstrated the critical function and mechanisms of LLDT-8 in osteoclast inhibition, providing new ideas for the potential effectiveness of intervention RA with LLDT-8.

The chronic autoimmune disease multiple sclerosis (MS) now remains incurable. Paeoniflorin (PF), which is a monoterpene glucoside obtained from Paeonia lactiflora Pall, is recognized for neuroprotective and anti-inflammatory properties. However, the precise mechanism by which PF regulates MS is unclear. This work aims to elucidate the underlying mechanisms of PF in EAE, a well established animal model of MS, and to discover the target proteins that PF directly acts on. Our results revealed that PF administration can significantly attenuate the clinical symptoms of EAE and alleviate the central nervous system (CNS) inflammatory environment by inhibiting M1-type microglia/macrophages. Mechanistically, PF was found to directly interact with the glycolytic enzyme α-enolase (ENO1), inhibiting its enzymatic activity and expression to impair glucose metabolism, thereby suppressing microglia/macrophage M1 polarization and ameliorating CNS inflammation. Significantly, Eno1 knockdown in microglia/macrophages diminished their pro-inflammatory phenotype, while treatment with ENOBlock or the specific knockout of Eno1 in microglia led to EAE remission, underscoring the critical role of ENO1 in EAE progression. This study uncovers the molecular mechanism of PF in treating EAE, linking the anti-inflammatory property of PF to the glucose metabolism process, which will broaden the prospective applications of PF.

"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".

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by a high level of autoantibody production. T follicular helper (Tfh) cells and B cells participate in the development of SLE. Several studies have shown that CXCR3+ cells are increased in SLE patients. However, the mechanism through which CXCR3 influences lupus development remains unclear. In this study, we established lupus models to determine the role of CXCR3 in lupus pathogenesis. The concentration of autoantibodies was detected using the enzyme-linked immunosorbent assay (ELISA), and the percentages of Tfh cells and B cells were measured using flow cytometry. RNA sequencing (RNA-seq) was performed to detect the differentially expressed genes in CD4+ T cells from wild-type (WT) and CXCR3 knock-out (KO) lupus mice. Migration of CD4+ T cells in spleen section was assessed using immunofluorescence. CD4+ T cell function in helping B cells produce antibodies was determined using a co-culture experiment and supernatant IgG ELISA. Lupus mice were treated with a CXCR3 antagonist to confirm the therapeutic effects. We found that the expression of CXCR3 was increased in CD4+ T cells from lupus mice. CXCR3 deficiency reduced autoantibody production with decreased proportions of Tfh cells, germinal center (GC) B cells, and plasma cells. Expression of Tfh-related genes was downregulated in CD4+ T cells from CXCR3 KO lupus mice. Migration to B cell follicles and T-helper function of CD4+ T cells were reduced in CXCR3 KO lupus mice. CXCR3 antagonist AMG487 decreased the level of serum anti-dsDNA IgG in lupus mice. We clarify that CXCR3 may play an important role in autoantibody production by increasing the percentages of aberrant activated Tfh cells and B cells and promoting the migration and T-helper function of CD4+ T cells in lupus mice. Thus, CXCR3 may be a potential target for lupus therapy.

目的:探讨LLDT-8对类风湿关节炎骨破坏的作用机制.方法:收集取类风湿关节炎患者的外周血和关节滑液,分离外周血单个核细胞(PBMC)和关节滑液单个核细胞(SFMC).PBMC用RANKL和M-CSF分别诱导后加入不同浓度LLDT-8干预,于24、48、72 h和14 d进行抗酒石酸酸性磷酸酶染色,显微镜下对Trap(+)细胞计数,同时收集上清检测RANKL、RANK、OPG,比较组间差异.PBMC、SFMC用不同浓度LLDT-8干预后,用ELISA方法检测不同标本中RANKL、OPG、VEGF的分泌,另外用实时PCR检测不同标本PBMC中RANKL、OPG、VEGF、MMP-9的表达,以了解LLDT-8对骨破坏相关因子的影响.结果:LLDT-8减少PBMC向破骨样细胞的转化,促进破骨样细胞的凋亡;LLDT-8抑制VEGF的表达,上调OPG mRNA表达,同时下调VEGF、RANKL和MMP-9的表达.结论:LLDT-8促进破骨样细胞的凋亡,抑制OC的分化,降低RANKL/OPG比值,下调VEGF的表达,延缓骨破坏.

Protein arginine methyltransferase 5 (PRMT5) participates in the symmetric dimethylation of arginine residues of proteins and contributes to a wide range of biological processes. However, how PRMT5 affects the transcriptional and epigenetic programs involved in the establishment and maintenance of T cell subset differentiation and roles in antitumor immunity is still incompletely understood. In this study, using single-cell RNA and chromatin immunoprecipitation sequencing, we found that mouse T cell-specific deletion of PRMT5 had greater effects on CD8(+) than CD4(+) T cell development, enforcing CD8(+) T cell differentiation into Klrg1(+) terminal effector cells. Mechanistically, T cell deficiency of PRMT5 activated Prdm1 by decreasing H4R3me2s and H3R8me2s deposition on its loci, which promoted the differentiation of Klrg1(+)CD8(+) T cells. Furthermore, effector CD8(+) T cells that transited to memory precursor cells were decreased in PRMT5-deficient T cells, thus causing dramatic CD8(+) T cell death. In addition, in a mouse lung cancer cell line-transplanted tumor mouse model, the percentage of CD8(+) T cells from T cell-specific deletion of PRMT5 mice was dramatically lost, but CD8(+)Foxp3(+) and CD8(+)PDL1(+) regulatory T cells were increased compared with the control group, thus accelerating tumor progression. We further verified these results in a mouse colon cancer cell line-transplanted tumor mouse model. Our study validated the importance of targeting PRMT5 in tumor treatment, because PRMT5 deficiency enforced Klrg1(+) terminal CD8(+) T cell development and eliminated antitumor activity.

Rationale: Protein arginine methyltransferase 5 (PRMT5) is an oncogene that promotes tumor cell proliferation, invasion and metastasis. However, the underlying mechanisms by which PRMT5 contributes to the progression of cervical cancer and especially the tumor microenvironment remain poorly understood. Methods: PRMT5 expression level was analyzed by Q-PCR, western blot, immunohistochemistry, and TCGA database. The role of PRMT5 in tumor growth was observed by transplanted tumor models, and the function of T cells in tumor microenvironment and in vitro co-culture system was investigated through flow cytometry. The transcriptional regulation of PRMT5 was analyzed using luciferase reporter and chromatin immunoprecipitation (ChIP) assay. The therapeutic effect of PRMT5 inhibitor was evaluated in a cervical cancer cell line transplanted tumor model. Results: We observed that the mRNA and protein expression levels of PRMT5 were increased in cervical cancer tissues, and the high expression of PRMT5 was associated with poor outcomes in cervical cancer patients. The absence of PRMT5 significantly inhibited tumor growth in a cervical cancer transplanted tumor model, and importantly, PRMT5 absence in tumors led to increase the number and enhance the function of tumor infiltrating T cells. Mechanistically, PRMT5 enhanced the transcription of STAT1 through symmetric dimethylation of histone H3R2 and thus promoted PD-L1 expression in cervical cancer cells. Moreover, in an in vitro co-culture system, knockdown of PRMT5 in tumor cells could directly enhance the expression of IFN-γ, TNF-α and granzyme B in T cells. These results suggested that PRMT5 promoted the development of cervical cancer by the crosstalk between tumor cells and T cells. Furthermore, the PRMT5 inhibitor EPZ015666 treatment could suppress tumor growth in a cervical cancer transplanted tumor model. Conclusion: Our results clarify a new mechanism which PRMT5 knockdown in cervical cancer cells drives an antitumor function via reprogramming T cell-mediated response and regulating PD-L1 expression. Thus, our study highlights that PRMT5 may be a potential target for cervical cancer therapy.

Background PRMT5 is a type II protein arginine methyltransferase that methylates histone or non-histone proteins. Arginine methylation by PRMT5 has been implicated in gene transcription, ribosome biogenesis, RNA transport, pre-mRNA splicing and signal transduction. High expression of PRMT5 has been observed in various cancers and PRMT5 overexpression has been reported to improve cancer cell survival, proliferation, migration and metabolism and to inhibit cancer cell apoptosis. In addition, PRMT5 has been found to be required for cancer stem cell survival, self-renewal and differentiation. Several microRNAs have been shown to regulate PRMT5 expression. As PRMT5 has oncogene-like properties, several PRMT5 inhibitors have been used to explore their efficacy as potential drugs for different types of cancer, and three of them are now being tested in clinical trials. Conclusions In this review, we summarize current knowledge on the role of PRMT5 in cancer development and progression, including its functions and underlying mechanisms. In addition, we highlight the rapid development of PRMT5 inhibitors and summarize ongoing clinical trials for cancer therapy. By affecting both tumor cells and the tumor microenvironment, PRMT5 inhibitors may serve as effective anti-cancer agents, especially when combined with immune therapies.

Genome-wide association studies have identified >100 genetic risk factors for rheumatoid arthritis. However, the reported genetic variants could only explain less than 40% heritability of rheumatoid arthritis. The majority of the heritability is still missing and needs to be identified with more studies with different approaches and populations. In order to identify novel function SNPs to explain missing heritability and reveal novel mechanism pathogenesis of rheumatoid arthritis, 4 HLA SNPs (HLA-DRB1, HLA-DRB9, HLA-DQB1, and TNFAIP3) and 225 common SNPs located in miRNA, which might influence the miRNA target binding or pre-miRNA stability, were genotyped in 1,607 rheumatoid arthritis and 1,580 matched normal individuals. We identified 2 novel SNPs as significantly associated with rheumatoid arthritis including rs1414273 (miR-548ac, OR = 0.84, p = 8.26 × 10-4) and rs2620381 (miR-627, OR = 0.77, p = 2.55 × 10-3). We also identified that rs5997893 (miR-3928) showed significant epistasis effect with rs4947332 (HLA-DRB1, OR = 4.23, p = 0.04) and rs2967897 (miR-5695) with rs7752903 (TNFAIP3, OR = 4.43, p = 0.03). In addition, we found that individuals who carried 8 risk alleles showed 15.38 (95%CI: 4.69-50.49, p < 1.0 × 10-6) times more risk of being affected by RA. Finally, we demonstrated that the targets of the significant miRNAs showed enrichment in immune related genes (p = 2.0 × 10-5) and FDA approved drug target genes (p = 0.014). Overall, 6 novel miRNA SNPs including rs1414273 (miR-548ac, p = 8.26 × 10-4), rs2620381 (miR-627, p = 2.55 × 10-3), rs4285314 (miR-3135b, p = 1.10 × 10-13), rs28477407 (miR-4308, p = 3.44 × 10-5), rs5997893 (miR-3928, p = 5.9 × 10-3) and rs45596840 (miR-4482, p = 6.6 × 10-3) were confirmed to be significantly associated with RA in a Chinese population. Our study suggests that miRNAs might be interesting targets to accelerate understanding of the pathogenesis and drug development for rheumatoid arthritis.

The IL-7/IL-7R pathway plays a vital role in the immune system, especially in the inflammatory response. Monocytes/macrophages (osteoclast precursors) have been recently recognized as important participants in the osteoclastogenesis of rheumatoid arthritis (RA) patients. Here, we aimed to investigate the therapeutic potential of IL-7/IL-7R pathway in RA and to determine whether it could restrain osteoclastogenic functions and therefore ameliorate RA. Firstly, collagen-induced arthritis (CIA) mice were administered with IL-7Rα-target antibodies to assess their therapeutic effect on arthritis. We found that blockade of the IL-7/IL-7R pathway protected CIA mice from bone destruction in addition to inducing inflammatory remission, by altering the RANKL/RANK/OPG ratio and consequently decreasing osteoclast formation. To explore the effect and mechanism of this pathway, bone marrow cells were induced to osteoclasts and treated with IL-7, a STAT5 inhibitor or supernatants from T cells. The results showed that the IL-7/IL-7R pathway played a direct inhibitory role in osteoclast differentiation via STAT5 signalling pathway in a RANKL-induced manner. We applied flow cytometry to analyse the effect of IL-7 on T-cell RANKL expression and found that IL-7/IL-7R pathway had an indirect role in the osteoclast differentiation process by enhancing the RANKL expression on T cells. In conclusion, the IL-7/IL-7R pathway exhibited a dual effect on osteoclastogenesis of CIA mice by interacting with osteoimmunology processes and could be a novel therapeutic target for autoimmune diseases such as RA.

Rheumatoid arthritis (RA) is a complex systemic autoimmune disorder that primarily involves joints, further affects the life quality of patients, and has increased mortality. The pathogenesis of RA involves multiple pathways, resulting in some patients showing resistance to the existing drugs. Salubrinal is a small molecule compound that has recently been shown to exert multiple beneficial effects on bone tissue. However, the effect of Salubrinal in RA has not been clearly confirmed. Hence, we induced collagen-induced arthritis (CIA) in DBA/1J mice and found that Salubrinal treatment decreased the clinical score of CIA mice, inhibiting joint damage and bone destruction. Furthermore, Salubrinal treatment downregulated osteoclast number in knee joint of CIA in mice, and suppressed bone marrow-derived osteoclast formation and function, downregulated osteoclast-related gene expression. Moreover, Salubrinal treatment inhibited RANKL-induced NF-κB signaling pathway, and promoted P65 degradation through the ubiquitin-proteasome system, further restrained RANKL-induced osteoclastogenesis. This study explains the mechanism by which Salubrinal ameliorates arthritis of CIA in mice, indicating that Salubrinal may be a potential drug for RA, and expands the potential uses of Salubrinal in the treatment of bone destruction-related diseases.

思想政治工作贯穿教育教学全过程,立德树人是中心环节.根据免疫学专业课程特点,将课程思政融入免疫学教学是一种两者相得益彰的教学方式.本文总结了课程思政与免疫学教学融合的必要性,思考并探索了将课程思政融入免疫学教学的内容和方式,以实现全程育人和全方位育人的目标,培养卓越的医学生.

目的:观测小鼠磨牙加力不同时间点脾脏CD3+、CD8+、CD4+T淋巴细胞及Th细胞亚群的比例变化,分析正畸加力对免疫系统中T淋巴细胞变化的影响.方法:4周龄雄性C57BL/6小鼠随机分为1 d、3 d、5 d和7 d加力组以及相应时间的对照组.加力组向磨牙施加25 g力;对照组安置相同加力装置,力值为0.使用流式细胞术检测小鼠脾脏单个核细胞中CD3+比例、CD4+/CD3+比例、CD8+/CD3+比例及Th1,Th2,Th17细胞占CD4+细胞比例,以及CD3,CD4,CD8,IFN-γ,IL-4,IL-17荧光强度的变化.结果:与对照组相比,7 d实验组CD8+T细胞占CD3+T细胞的比例和Th1细胞占CD4+T淋巴细胞的比例显著增高(P<0.05);CD3,CD8,IFN-γ,IL-17平均荧光强度均显著增高(P<0.05);相反,CD4+T细胞占CD3+T淋巴细胞的比例显著降低(P<0.01).而1,3,5 d实验组与对照组间未见明显差异(P>0.05).结论:正畸加力过程可引起小鼠脾脏T细胞亚群占比和及其分泌的细胞因子发生变化.

Cannabinoids are widely studied as therapeutic agents for the treatment of various diseases. Among them, THC and CBD are two important phytocannabinoids which have served as structural templates for the design of synthetic analogs. In this study, we designed and synthesized a variety of novel cannabinoids based on the structural backbones of THC and CBD using the carbon-silicon switch strategy. A dimethyl silyl group was introduced as the tail group and two series of novel compounds were designed and synthesized, which showed a wide range of binding affinity for CB1 and CB2 receptors. Among them, compound 15b was identified as a non-selective CB1 and CB2 agonist and 38b as a selective agonist for the CB2 receptor. Preliminary screening showed that both compounds have improved metabolic stability than their carbon analogs and good in vivo pharmacokinetic profiles. Furthermore, both 15b and 38b significantly alleviated the phenotype of experimental autoimmune encephalomyelitis (EAE) in mice.

目的 通过检测乳腺癌患者腋窝可疑淋巴结细针穿刺(FNA)洗脱液中肿瘤标志物的浓度,为诊断淋巴结是否为乳腺癌转移淋巴结提供依据.方法 收集128例乳腺癌初发患者腋窝可疑淋巴结超声引导下的FNA洗脱液标本242份及配对血清标本128份.采用免疫化学发光法检测标本中肿瘤标志物癌胚抗原(CEA)、糖类抗原(CA)153及细胞角蛋白19片段(CYFRA21-1)的浓度.结合组织病理结果回顾性分析转移组和阴性组FNA洗脱液标志物浓度的差异.分析洗脱液和比值法3种肿瘤标志物检测乳腺癌转移的性能,并与超声引导下细针穿刺细胞学检查(US-FNAC)进行比较.结果 242份淋巴结标本中,70份为阴性,172份提示转移.转移淋巴结洗脱液中CEA、CA153和CYFRA 21-1检测浓度均显著高于阴性淋巴结(P<0.001).US-FNAC诊断的敏感性为83.7％,准确性为88.4％、特异性为100.0％.洗脱液中检测CEA、CA153和CYFRA21-1的诊断敏感性分别为50.1％、44.8％和95.3％,特异性分别为92.9％、91.4％和92.9％,准确性分别为62.8、58.3％和94.6％.比值法检测CEA、CA153和CYFRA21-1的诊断敏感性分别为47.7％、40.1％和94.2％,准确性分别为62.8％、57.4％和95.9％,特异性均为100.0％.US-FNAC联合比值法CYFRA21-1检测乳腺癌能将检测的敏感性提高到97.1％,准确性提高到97.9％.结论 在乳腺癌患者可疑淋巴结的诊断中,CYFRA21-1检测在洗脱液和比值法中的表现均优于CEA或CA153.US-FNAC联合比值法CYFRA21-1检测有助于提高检测敏感性和准确性,且保持特异性仍为100％.

Although CD4(+)CD45RA(-)Foxp3(l)degrees cytokine-secreting T cells (Fr.III cells) have been reported to be increased in systemic lupus erythematosus (SLE), their function and effects on response of B cells are still unclear. Here, we dissect how BACH2 regulates Fr.III cells function and promotes B-cell response in active SLE patients. We measured cytokines and BACH2 expression, and found that Fr.III cells from SLE patients produce much more inflammatory cytokines and were more able to promote B- cell proliferation, IgG, IgA, and TNF-alpha production than controls in a co-culture system. Fr.III cells expressed high levels of ICOS and CD154, but a low level of Tfr and BACH2, BACH2 expression was negatively correlated with SLE Disease Activity Index. Overexpressed of BACH2 in Fr.III cells, decreased cytokines expression and reduced B-cell response. Furthermore, we identified a reduction of H3K27ac level binding at the BACH2 locus in the SLE Fr.III cells and SLE serum stimulation decreased H3K27ac binding at the BACH2 locus, which could be restored using trichostatin A (TSA). In conclusion, BACH2 was associated with SLE disease activity, regulated the function of Fr.III cells, and promoted B-cells response. Targeting BACH2 may be a new immune intervention therapy of SLE.

RA是一种自身免疫性疾病,CD4+T细胞在其发生发展中发挥重要作用.研究通过分析CIA小鼠Salubrinal治疗组和对照组脾脏致病性细胞的增殖,Th亚群比例和相关转录因子表达的差异,以及关节病变部位炎症细胞浸润和细胞因子的变化,探索Salubrinal治疗CIA的作用机制.结果 显示,Salubrinal可以抑制脾脏中Ⅱ型胶原反应性CD4+T细胞的增殖,下调Th1和Th17比例以及转录因子T-bet和p-STAT3的表达,抑制关节病变部位炎症细胞浸润和IL-17与TNF-α的表达,提示Salubrinal通过调节Th1和Th17比例以及对抗原特异性CD4+T细胞增殖的抑制治疗CIA.本研究为RA的治疗提供了新思路.

Programmed death-1 (PD-1) is a cell surface receptor that functions as a T cell checkpoint and plays a central role in regulating T cell exhaustion. Binding of PD-1 to its ligand, programmed death-ligand 1 (PD-L1), activates downstream signaling pathways and inhibits T cell activation. Moreover abnormally high PD-L1 expression on tumor cells and antigen-presenting cells in the tumor microenvironment mediates tumor immune escape, and the development of anti-PD-1/PD-L1 antibodies has recently become a hot topic in cancer immunotherapy. Here, we review the structure of PD-1 and PD-L1, the function of the PD-1/PD-L1 signaling pathway, the application of PD-1 or PD-L1 monoclonal antibodies and future directions for anti-PD-1/PD-L1 antibodies with combination therapies. Cancer immunotherapy using PD-1/PD-L1 immune checkpoint blockade may require more studies, and this approach may be curative for patients with many types of cancer in the future.

Backgrounds To explore the expression and functions of the tripartite motif-containing protein 21 (TRIM21) in oral lichen planus(OLP) lesions. Methods Paraffin sections of buccal mucosa samples from 15 cases of reticular oral lichen planus (OLP) patients and 10 healthy controls were used for immunohistochemistry to determine expression and distribution of TRIM21. Buccal mucosae from 11 OLP patients and seven healthy controls were analyzed by qPCR to quantify its gene expression. Peripheral blood mononuclear cells and CD3+ cells from four pairs of age- and sex-matched OLP patients and healthy controls were isolated for immunocytochemistry and culture. Following lentivirus-mediated overexpression of TRIM21 gene in CD3+ cells, CCK-8 was applied to evaluate cell proliferation. Cytokines including IL-2, IL-4, IL-5, IL-6, IL-10, TNF-alpha, and IFN-gamma in the supernatants were measured by the cytometric bead array and verified by ELISA. Results A larger number of TRIM21-positive cells infiltrating the lamina propria were observed in OLP lesions by immunohistochemistry than those of healthy controls. Significantly higher transcription of TRIM21 was revealed by qPCR. TRIM21 overexpression in CD3+ cells significantly enhanced the proliferation and IL-6 secretion in CD3+ cells from 12 to 72 hours. Conclusion Overexpressed TRIM21 in OLP may be a primary proinflammatory molecule rather than a secondary and inducible regulatory factor in immunopathogenesis of OLP.

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.