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Enhanced therapeutic effects of apoptotic cell-conditioned mesenchymal stem cells in lupus-prone MRL/lpr mice

Rheumatology & Autoimmunity 2024年第2期4卷 90-98页

作者： Zhuoya Zhang Yiyuan Cui Saisai Huang Weilin Liu Chen Chen xuebing feng Dandan Wang Lingyun Sun Department of Rheumatology and Immunology Nanjing Drum Tower HospitalAffiliated Hospital of Medical SchoolNanjing UniversityNanjingJiangsuChina Department of Rheumatology and Immunology Nanjing Drum Tower HospitalNanjing Drum Tower Hospital Clinical College of Traditional Chinese and Western MedicineNanjing University of Chinese MedicineNanjingJiangsuChina Department of Nutrition Nanjing Drum Tower HospitalAffiliated Hospital of Medical SchoolNanjing UniversityNanjingJiangsuChina

Background::Apoptotic cell-conditioned mesenchymal stem cells (AC-MSCs) exhibit stronger T cell suppressive ability via cyclooxygenase 2 (COX2)/prostaglandin E2 (PGE2);however, whether AC-MSCs exhibit enhanced therapeutic effects on systemic lupus erythematosus (SLE) remains unknown. Methods: Splenocytes from MRL/MPJ-Fas lpr (MRL /lpr) mice were cocultured with AC-MSCs, and the proportion of plasma cells was determined by flow cytometry. MSCs, AC-MSCs, COX2 knockdown MSCs, and COX2 knockdown AC-MSCs were infused into MRL/ lpr mice ( n = 10/group). Survival rates and lupus symptoms, including proteinuria, kidney injury, renal immune complex deposition, and autoantibody production, were assessed. Additionally, the number of plasma cells and serum levels of inflammatory cytokines were measured. Results::The AC-MSCs significantly inhibited plasma cells via PGE2 after 24 h coculture in vitro, whereas MSCs did not. In the MRL /lpr mice, AC-MSC treatment led to a significantly higher survival rate than phosphate-buffered saline (PBS) treatment (90% vs. 50%, p < 0.05). Moreover, AC-MSC infusion decreased urine protein levels as early as 1 week after administration (0.89 ± 0.55 mg/mL vs. 1.59 ± 0.60 mg/mL, p < 0.05, compared with PBS treatment). Administration of both MSCs and AC-MSCs reduced renal immunoglobulin G and complement C3 deposition, whereas COX2 knockdown MSCs and COX2 knockdown AC-MSCs did not. Serum anti-dsDNA antibody levels in AC-MSC-treated mice significantly decreased (0.40 ± 0.25 vs. 0.99 ± 0.58, p < 0.05), compared with PBS treatment, as well as the number of plasma cells in both the spleen ([2.14 ± 1.05] × 10^(6) vs. [8.02 ± 4.01] × 10^(6), p < 0.01) and renal-draining lymph nodes ([0.78 ± 0.68] × 10^(6) vs. [2.49 ± 1.45] × 10^(6), p < 0.05). Additionally, AC-MSCs inhibited the production of inflammatory cytokines, including interleukin-21, tumor necrosis factor-alpha, and monocyte chemoattractant protein-1. Conclusions::AC-MSCs enhanced the therapeutic

关键词： apoptotic cells mesenchymal stem cells precondition prostaglandin E2 systemic lupus erythematosus

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中国系统性红斑狼疮的诊治现状及挑战

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诊断学理论与实践 2024年第3期23卷 257-262页

作者：张昕赵盛楠冯学兵南京大学医学院附属鼓楼医院风湿免疫科江苏南京210008

系统性红斑狼疮(systemic lupus erythematosus,SLE)是一种以多系统损害为特征的自身免疫性疾病。我国SLE患者基数大、病情重、缓解率低、复发率高、感染风险高。2023年,研究显示,全球有SLE患者约341万例,其中我国患者达70~100万例,总... 详细信息

系统性红斑狼疮(systemic lupus erythematosus,SLE)是一种以多系统损害为特征的自身免疫性疾病。我国SLE患者基数大、病情重、缓解率低、复发率高、感染风险高。2023年,研究显示,全球有SLE患者约341万例,其中我国患者达70~100万例,总数位居首位。我国SLE患者的平均发病年龄为30.7岁,女性与男性的发病比为12∶1;脏器受累更为多见,其中肾脏受累率为45.02%,血液系统受累率为37.2%,均显著高于欧洲SLE患者(肾脏受累率为27.9%,血液系统受累率为18.2%)。我国SLE患者疾病治疗的临床缓解率为2.47%,而国际报道的缓解率达22.9%。当前,我国SLE患者的短期生存率基本与国际一致(5年可达94%),但长期生存率仍不容乐观,呈断崖式下跌,25~30年生存率仅为30%。我国有84.13%的SLE患者使用糖皮质激素治疗,而国际上SLE患者的糖皮质激素使用率仅为42.6%,感染是我国SLE患者死亡的首位原因,而西方国家SLE患者死因则主要为心血管疾病和肿瘤。随着新分类标准的提出、达标治疗理念的引入、新型治疗方法的运用,将大大推动SLE诊疗的发展,有望进一步改善我国患者的预后。

关键词：系统性红斑狼疮分类标准达标治疗生物制剂

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The regulation of the Treg/Th 17 balance by mesenchyma stem cells in human systemic lupus erythematosus

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Cellular & Molecular Immunology 2017年第5期14卷 423-431页

作者： Dandan Wang Saisai Huang Xinran Yuan Jun Liang Renju Xu Genhong Yao xuebing feng Lingyun Sun Department of Rheumatology and Immunology The Affiliated Drum Tower Hospital of Nanjing University Medical School Nanjing China

Background and objective： Umbilical cord （UC）-derived mesenchymal stem cells （MSCs） have shown immunoregulation of various immune cells. The aim of this study was to investigate the mechanism of UC MSCs in the regulation of peripheral regulatory T cells （Treg） and T helper 17 （Th17） cells in patients with systemic lupus erythematosus （SLE）. Methods： Thirty patients with active SLE, refractory to conventional therapies, were given UC MSCs infusions. The percentages of peripheral blood CD4＋CD25＋Foxp3＋ regulatory T cells （Treg） and CD3＋CD8-1L17A＋ Th17 cells and the mean fluorescence intensities （MFI） of Foxp3 and IL- 17 were measured at I week, I month, 3 months, 6 months, and 12 months after MSCs transplantation （MSCT）. Serum cytokines, including transforming growth factor beta （TGF-β）, tumor necrosis factor alpha （TNF-α）, interleukin 6 （IL-6）, and IL-17A were detected using ELISA. Peripheral blood mononuclear cells from patients were collected and co-cultured with UC MSCs at ratios of 1：1, 10：1, and 50：1, respectively, for 72 h to detect the proportions of Treg and Th17 cells and the MFIs of Foxp3 and IL-17 were determined by flow cytometry. The cytokines in the supernatant solution were detected using ELISA. Inhibitors targeting TGF-β, IL-6, indoleamine 2,3-dioxygenase （IDO）, and prostaglandin E2 were added to the co-culture system, and the percentages of Treg and Th17 cells were observed. Results： The percentage of peripheral Treg and Foxp3 MFI increased 1 week, 1 month, and 3 months after UC MSCs transplantation, while the Th17 proportion and MFI of IL-17 decreased 3 months, 6 months, and 12 months after the treatment, along with an increase in serum TGF-β at I week, 3 months, and 12 months and a decrease in serum TNF-a beginning at I week. There were no alterations in serums IL-6 and IL-17A before or after MSCT. In vitro studies showed that the UC MSCs dose-dependently up-regulated peripheral Treg proportion in SLE patients, which was not depended on cell-cell co

关键词： mesenchymal stem cells regulatory T cells systemic lupus erythematosus T helper 17 cells umbilical cord

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MicroRNA-663 induces immune dysregulation by inhibiting TGF-β1 production in bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus

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Cellular & Molecular Immunology 2019年第3期16卷 260-274页

作者： Linyu Geng Xiaojun Tang Kangxing Zhou Dandan Wang Shiying Wang Genhong Yao Weiwei Chen Xiang Gao Wanjun Chen Songtao Shi Nan Shen xuebing feng Lingyun Sun Department of Rheumatology and Immunology The Affiliated Drum Tower Hospital of Nanjing University Medical SchoolNanjing 210008China Key Laboratory of Model Animal for Disease Study Model Animal Research CenterNanjing UniversityNanjing 210000China Mucosal Immunology Section OPCBNational Institute of Dental and Craniofacial ResearchNational Institutes of HealthBethesda 20892-2190MDUSA Department of Anatomy and Cell Biology School of Dental MedicineUniversity of PennsylvaniaPhiladelphia 19104-6004PAUSA Joint Molecular Rheumatology Laboratory of the Institute of Health Sciences and Shanghai Renji Hospital Shanghai Institutes for Biological SciencesChinese Academy of Sciencesand Shanghai Jiaotong University School of MedicineShanghaiChina

Mesenchymal stem cells(MSCs)are critical for immune *** several microRNAs(miRNAs)have been shown to participate in autoimmune pathogenesis by affecting lymphocyte development and function,the roles of miRNAs in MSC dysfunction in autoimmune diseases remain ***,we show that patients with systemic lupus erythematosus(SLE)display a unique miRNA signature in bone marrow-derived MSCs(BMSCs)compared with normal controls,among which miR-663 is closely associated with SLE disease ***-663 inhibits the proliferation and migration of BMSCs and impairs BMSC-mediated downregulation of follicular T helper(Tfh)cells and upregulation of regulatory T(Treg)cells by targeting transforming growth factorβ1(TGF-β1).MiR-663 overexpression weakens the therapeutic effect of BMSCs,while miR-663 inhibition improves the remission of lupus disease in MRL/lpr ***,miR-663 is a key mediator of SLE BMSC regulation and may serve as a new therapeutic target for the treatment of lupus.

关键词： immune dysregulation mesenchymal stem cells miR-663 systemic lupus erythematosus transforming growth factorβ1

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Transplantation of Human Bone Marrow Mesenchymal Stem Cell Ameliorates the Autoimmune Pathogenesis in MRL/lpr Mice

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Cellular & Molecular Immunology 2008年第6期5卷 417-424页

作者： Kangxing Zhou Huayong Zhang Ouyang Jin xuebing feng Genhong Yao Yayi Hou Lingyun Sun Department of Rheumatology and Immunology The Affiliated DrumTower Hospital of Nanjing University Medical School 321 ZhongshanRoad Nanjing Jiangsu 210008 China Immunology Laboratory Nanjing University Medical School 22 HankouRoad Nanj ing Jiangsu 2 10093 China These authors contributed equally to this work

Recent evidence indicates that mesenchymal stem cells （MSC） possess immunosuppressive properties both in vitro and in vivo. We previously demonstrated the functional abnormality of bone marrow derived MSC in patients with systemic lupus erythematosus （SLE）. In this study, we aimed to investigate whether transplantation of human bone marrow derived MSC affects the autoimmune pathogenesis in MRL/Ipr mice. We found that human MSC from healthy donors reduced the proliferation of T lymphocytes from MRL/Ipr mice in a dose-dependent fashion. Two weeks after in vivo transfer of MSC, we detected significantly reduced serum levels of anti ds-DNA antibodies and 24 hour proteinuria in MRL/Ipr mice as compared with control groups without MSC transplantation. Moreover, flow cytometric analysis revealed markedly reduced number of CD4＋ T cells while increased Thl subpopulation in MSC group and MSC ＋ CTX group when compared with controls. Histopathological examination showed significantly reduced renal pathology in MSC-treated mice. Immunohistochemical studies further revealed reduced expression of TGF-~, FN, VEGF and the deposition of complement C3 in renal tissue after MSC and MSC ＋ CTX treatment. Taken together, we have demonstrated that transplantation of human MSC can significantly inhibit the autoimmune progression in MRL/Ipr mice. Cellular ＆ Molecular Immunology. 2008;5（6）：417-424.

关键词： systemic lupus erythematosus mesenchymal stem cell transplantation

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MSCs relieve SLE by modulation of Th17 cells through MMPs–CCL2–CCR2–IL-17 pathway

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Rheumatology & Autoimmunity 2021年第1期1卷 30-39页

作者： Bo Jiang Genhong Yao Xiaojun Tang Xi Yang xuebing feng Department of Rheumatology and Immunology The Affiliated Drum Tower Hospital of Nanjing University Medical SchoolNanjingJiangsuPeople's Republic of China

Objective:To explore the modulation of mesenchymal stem cells(MSCs)on T helper 17(Th17)cells in systemic lupus erythematosus(SLE)and underlying ***:The concentration of matrix metalloproteinases(MMPs),CC chemokine ligand-2(CCL2),and interleukin-17(IL-17)in the serum of SLE patients and mice were detected by enzyme-linked immunosorbent *** expression of CCR2 and IL-17 of T lymphocytes were determined by flow *** effects of MSCs on Th17 cells were analyzed in lupus mice and coculture system in ***:The levels of MMPs,CCL2,IL-17,CCR2,and percentages of Th17 cells were significantly increased in SLE *** molecules and numbers of Th17 cells were downregulated by umbilical cord-derived MSCs(UC-MSCs)which relieve SLE ***2 neutralizing antibody blocked the effects of MSCs on Th17 *** reversed the function of ***:The beneficial effects of MSCs on SLE patients rely on secreting MMPs,which reverse the activity of CCL2 to inhibit Th17 cells,suggesting the crucial MSCs–MMP–CCL2–CCR2–Th17–IL-17 pathway in SLE.

关键词： CC chemokine ligand-2 mesenchymal stem cells systemic lupus erythematosus T helper 17 cells

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Flexible bidirectional pulse charging regulation achieving long-life lithium-ion batteries

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Journal of Energy Chemistry 2024年第9期96卷 59-71页

作者： Xiaodong Xu Shengjin Tang xuebing Han Languang Lu Yudi Qin Jiuyu Du Yu Wu Yalun Li Chuanqiang Yu Xiaoyan Sun Xuning feng Minggao Ouyang Rocket Force University of Engineering Xi’an 710025ShaanxiChina State Key Laboratory of Intelligent Green Vehicle and Mobility School of Vehicle and MobilityTsinghua UniversityBeijing 100084China School of Materials Science and Engineering Beijing Institute of TechnologyBeijing 100081China

Typical application scenarios,such as vehicle to grid(V2G)and frequency regulation,have imposed significant long-life demands on lithium-ion ***,we propose an advanced battery life-extension method employing bidirectional pulse charging(BPC)*** traditional constant current charging methods,BPC strategy not only achieves comparable charging speeds but also facilitates V2G frequency regulation *** significantly enhances battery cycle ampere-hour throughput and demonstrates remarkable life extension *** this interesting conclusion,adopting model identification and postmortem characterization to reveal the life regulation mechanism of BPC:it mitigates battery capacity loss attributed to loss of lithium-ion inventory(LLI)in graphite anodes by intermittently regulating the overall battery voltage and anode potential using a negative charging ***,from the perspective of internal side reaction,the life extension mechanism is further revealed as inhibition of solid electrolyte interphase(SEI)and lithium dendrite growth by regulating voltage with a bidirectional pulse current,and a semi-empirical life degradation model combining SEI and lithium dendrite growth is developed for BPC scenarios health management,the model parameters are identified by genetic algorithm with the life simulation exhibiting an accuracy exceeding 99%.This finding indicates that under typical rate conditions,adaptable BPC strategies can extend the service life of LFP battery by approximately 123%.Consequently,the developed advanced BPC strategy offers innovative perspectives and insights for the development of long-life battery applications in the future.

关键词： Lithium-ion battery Long-life regulation Bidirectional pulse charging Mechanism identification

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Mizoribine versus mycophenolate mofetil or intravenous cyclophosphamide for induction treatment of active lupus nephritis

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Chinese Medical Journal 2014年第21期127卷 3718-3723页

作者： feng xuebing Gu Fei Chen Weiwei Liu Yan Wei Hua Liu Lin Yin Songlou Da Zhanyun Sun Lingyun Department of Rheumatology and Immunology Affiliated Drum Tower Hospital of Nanjing University Medical School Nanjing Jiangsu 210008 China Department of Rheumatology Huai'an First People's Hospital Nanjing Medical University Huai'an Jiangsu 223300 China Department of Rheumatology and Immunology Northern Jiangsu People's Hospital Yangzhou Jiangsu 225001 China Department of Rheumatology and Immunology Central Hospital of Xuzhou City Xuzhou Jiangsu 221009 China Department of Rheumatology and Immunology Affiliated Hospital of Xuzhou Medical College Xuzhou Jiangsu 221002 China Department of Rheumatology and Immunology Affiliated Hospital of Nantong University Nantong Jiangsu 226001 China

Background Lupus nephritis （LN） is one of the most serious manifestations of systemic lupus *** there have been substantial improvements in LN treatment over the last decade,the outcome remains unoptimistic in a considerable percentage of *** aim of this study was to evaluate the efficacy and safety of mizoribine （MZR）,a novel selective inhibitor of inosine monophosphate dehydrogenase,as induction treatment for active LN in comparison with mycophenolate mofetil （MMF） and intravenous cyclophosphamide （CYC）.Methods Ninety patients with active LN were *** patients were given MZR orally at the dose of 300 mg every other *** patients took MMF at 2 g per day in two divided *** patients received CYC intravenously 0.5 g every 2 *** effects and adverse events （AEs） were evaluated at the end of 24-week *** analysis of variance （ANOVA） followed by Dunn＇s test was applied to compare the difference among the *** comparing categorical data between two groups,χ^2 test was *** Early responses at week 12 were achieved by 73.3%,90.0%,and 96.7% in MZR,MMF,and CYC groups,*** was no significant difference in the complete remission rates （22.7%,24.0%,and 25.0%,respectively） or overall response rates （68.2%,72.0%,and 75.0%,respectively） among the three groups at week *** most prominent drop-down of Systemic Lupus Erythematosus Disease Activity Index scores was observed in MMF or CYC group,and the decline of health assessment questionnaire scores in MZR or MMF group was more prominent than that in the CYC group at week *** complement 3 （C3） or C4 levels were elevated in all groups after the *** was more effective in inhibiting anti-double-stranded DNA antibody,while MZR was more effective in inhibiting antinuclear *** incidences of AEs in patients treated with CYC were significantly higher than those in patients treated with MZR or MMF （24.2% for

关键词： mizoribine mycophenolate mofetil cyclophosphamide lupus nephritis

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嗜酸性粒细胞相关核糖核酸酶A家族成员2参与狼疮小鼠肾脏损害的机制研究

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中华风湿病学杂志 2024年第9期28卷 648-655,I0003页

作者：郑媛媛唐小军章雅琦米合热阿依·阿布都克尤木朱延通李文静冯学兵南京中医药大学鼓楼临床医学院、南京鼓楼医院风湿免疫科南京210008 南京大学医学院附属鼓楼医院风湿免疫科南京210008

目的通过嗜酸性粒细胞相关核糖核酸酶A家族成员2(Ear2)髓系细胞条件性敲除小鼠探讨其在狼疮发病中的作用及参与肾脏损害的可能机制。方法利用规律间隔成簇短回文重复序列关联基因9(CRISP/Cas9)技术构建Ear2髓系细胞条件性敲除小鼠模型,... 详细信息

目的通过嗜酸性粒细胞相关核糖核酸酶A家族成员2(Ear2)髓系细胞条件性敲除小鼠探讨其在狼疮发病中的作用及参与肾脏损害的可能机制。方法利用规律间隔成簇短回文重复序列关联基因9(CRISP/Cas9)技术构建Ear2髓系细胞条件性敲除小鼠模型,应用PCR鉴定小鼠基因型。实验分为3组:CKO+R848组,对照+R848组,对照组。R848处理敲除鼠和同型对照鼠,评估小鼠狼疮样病变发生情况。荧光实时定量PCR法检测小鼠肾脏中Toll样受体(TLR)7/8和炎症因子的表达。流式细胞术检测狼疮小鼠肾脏中巡逻单核细胞(PMOs)的比例,免疫荧光法分析Ear2和PMOs在肾组织中的空间分布。R848体外刺激条件性敲除(CKO)和对照小鼠的骨髓细胞,流式细胞术检测PMOs比例变化。采用单因素方差分析对3组进行比较。结果髓系细胞条件敲除Ear2小鼠构建后PCR显示基因型为Lyz2 ki/wtEar2 fl/fl,敲除鼠骨髓细胞中Ear2 mRNA水平显著下调[(1.03±0.26)与(0.22±0.15),t=6.65,P<0.001]。与对照+R848组相比,CKO+R848组小鼠狼疮表型得到改善,生存率有上升趋势(6/10与7/8,χ^(2)=1.51,P=0.220),病理结果提示CKO+R848组小鼠肾脏病变减轻。CKO+R848小鼠肾脏组织中TLR7表达水平降低[(1.02±0.09)与(0.53±0.04),t=5.13,P=0.003],同时PMOs浸润减少[(62.00±3.38)%与(52.36±0.68)%,t=2.80,P=0.023],免疫荧光结果显示Ear2和PMOs在肾组织共定位。体外R848刺激引起对照组PMOs比例的增多[(3.99±0.59)%与(33.48±1.38)%,t=-33.84,P<0.001],敲除Ear2可以抑制TLR7引起的PMOs的增多[(14.33±1.72)%与(16.10±1.44)%,t=-1.37,P=0.220]。结论Ear2条件性敲除可减轻狼疮小鼠发病,尤其是肾脏损害。其机制与抑制TLR7通路激活、减少PMOs局部浸润有关。

关键词：红斑狼疮,系统性 Toll样受体7 单核细胞嗜酸性粒细胞相关核糖核酸酶A家族成员2

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Targeted treatment strategy for patients with severe pulmonary hypertension secondary to connective tissue diseases

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Rheumatology & Autoimmunity 2023年第3期3卷 157-165页

作者： Bo Jiang Chun Wang Yansheng Jin Guanjun Guo Kangxing Zhou xuebing feng Genhong Yao Department of Rheumatology and Immunology The Affiliated Drum Tower Hospital of Nanjing University Medical SchoolNanjingJiangsuChina Department of Rheumatology and Immunology Wuzhong People's HospitalSuzhouJiangsuChina Department of Cardiac Ultrasound The Affiliated Drum Tower Hospital of Nanjing University Medical SchoolNanjingJiangsuChina

Background:Pulmonary arterial hypertension(PAH)associated with connective tissue diseases(CTD)(CTD-PAH)remains a difficult challenge in clinical *** aimed to evaluate the effects of targeted vasodilators in patients with severe ***:The data of 53 patients with severe CTD-PAH hospitalized at the Department of Rheumatology and Immunology,The Affiliated Drum Tower Hospital of Nanjing University Medical School,were retrospectively *** were followed up for an average of 2 years to track their *** efficacy of treatment and the survival rate of patients with severe CTD-PAH were ***:Among the causes of severe CTD-PAH,systemic lupus erythematosus(SLE)was the most common(39.6%),and the age at onset in patients with SLE-PAH was younger than that of patients with other *** was more effective than sildenafil in reducing pulmonary artery pressure,improving cardiac function,and increasing survival *** therapy with targeted vasodilators significantly improved the prognosis of patients with severe CTD-PAH compared with ***:Patients with severe CTD-PAH should be treated early with targeted *** this study,bosentan was superior to *** treatment might be an option for severe CTD-PAH.

关键词： connective tissue disease pulmonary arterial hypertension systemic lupus erythematosus targeted vasodilator

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