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A novel reporter mouse line for studying alveolar macrophages

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Science China(Life Sciences) 2023年第11期66卷 2527-2542页

作者： Xiaoyun Zhao Liang Li Hongxiang Sun Xiaoli Jiang Wenjuan Bai ningbo wu Jing Wang Bing Su Shanghai Institute of Immunology Department of Immunology and Microbiologyand the Ministry of Education Key Laboratory of Cell Death and DifferentiationShanghai Jiao Tong University School of MedicineShanghai 200025China Center for Immune-Related Diseases at Shanghai Institute of Immunology Department of GastroenterologyRuijin HospitalShanghai Jiao Tong University School of MedicineShanghai 200025China Shanghai Jiao Tong University School of Medicine—Yale Institute for Immune Metabolism Shanghai Jiao Tong University School of MedicineShanghai 200025China Key Laboratory of Molecular Radiation Oncology of Hunan Province Xiangya HospitalCentral South UniversityChangsha 410008China

Alveolar macrophages(AMs)are self-maintained immune cells that play vital roles in lung homeostasis and *** reporter mice and culture systems have been established for studying macrophages,an accurate and specific reporter line for alveolar macrophage study is still not *** we reported a novel Rspo1-tdTomato gene reporter mouse line that could specifically label mouse AMs in a cell-intrinsic *** this reporter system,we visualized the dynamics of alveolar macrophages intravitally under steady state and characterized the alveolar macrophage differentiation under in vitro *** performing ATAC-seq,we found that insertion of the tdTomato cassette in the Rspo1 locus increased the accessibility of a PPARE motif within the Rspo1 locus and revealed a potential regulation by key transcription factor PPAR-γfor alveolar macrophage differentiation in vitro and in ***,perturbation of PPAR-γby its agonist rosiglitazone or inhibitor GW9662 resulted in corresponding alteration of tdTomato expression in alveolar macrophages together with the transcription of PPAR-γdownstream target ***,global transcriptomic analyses of AMs from the wild type mice and the Rspo1-tdTomato mice showed comparable gene expression profiles,especially those AM-specific genes,confirming that the insertion of the tdTomato cassette in the Rspo1 locus does not impact the cell identity and biological function of AMs under normal *** together,our study provides an alternative tool for in vivo and in vitro labeling of alveolar macrophages with high specificity which could also be utilized as an indicator of PPAR-γactivity for future development of PPAR-γspecific targeting drugs.

关键词： alveolar macrophage gene reporter mice PPAR-γ R-spondin 1 ATAC-seq

scRNA-seq profiling of neonatal and adult thymus-derived CD4+ T cells by a T cell origin-time tracing model

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Journal of Molecular Cell Biology 2022年第12期14卷 1-16页

作者： Yuheng Han Xinxing Ouyang Yao Chen Shujing Lai Hongxiang Sun ningbo wu Chun Ruan Limin Lu Bing Su Shanghai Institute of Immunology Department of Immunology and Microbiologyand the Ministry of Education Key Laboratory of Cell Death and DifferentiationShanghai Jiao Tong University School of MedicineShanghai 200025China Department of Tumor Biology Shanghai Chest HospitalShanghai Jiao Tong University School of MedicineShanghai 200030China Center for Human Translational Immunology at Shanghai Institute of Immunology Ruijin HospitalShanghai Jiao Tong University School of MedicineShanghai 200025China Shanghai Jiao Tong University School of Medicine–Yale Institute for Immune Metabolism Shanghai Jiao Tong University School of MedicineShanghai 200025China Key Laboratory of Molecular Radiation Oncology of Hunan Province Xiangya HospitalCentral South UniversityChangsha 410008China

It is well documented that the neonatal thymus-derived (neonatal-TD) regulatory T cells (Treg) are essential to prevent lethal autoimmune diseases and allergies, and neonatal and adult thymus possesses distinct output potentials for naïve T cells, including Treg. However, the molecular features and detailed functional differences between neonatal-TD and adult thymus-derived (adult-TD) T cells in terms of their ability to maintain immune homeostasis during long-term environmental influences are still largely unknown, partially due to the lack of appropriate animal models to precisely trace these cells at specific time points. In this study, neonatal-TD and adult-TD CD4+ T cells from the spleen and Peyer's patches were traced for 9 weeks by a T cell origin-time tracing mouse model and analysed by single-cell RNA sequencing. More Treg but fewer naïve T cells were found in neonatal-TD CD4+ T cells from both tissues than those from adult-TD counterparts. Interestingly, the neonatal-TD Treg in both the spleen and Peyer's patches exhibited augmented expression of Foxp3, Gata3, Ctla4, Icos, Il2ra, Tgfb1, and Nrp1, as well as enriched Gene Ontology terms like T cell activation and tolerance induction, indicating an enhanced immunosuppressive function. These results were further confirmed by flow cytometry analysis and in vitro immune suppression assays. Flow cytometry also revealed a significantly higher proportion of neonatal-TD Treg in total Treg than that of adult-TD counterparts, suggesting the longer lifespan of neonatal-TD Treg. To investigate the intrinsic features of neonatal-TD and adult-TD CD4+ T cells, a shortened tracing time was performed. Surprisingly, the neonatal-TD and adult-TD CD4+ T cells had similar proportions of Treg and did not exhibit significant differences in Foxp3, Gata3, Ctla4, Icos, Il2ra, and Tgfb1 expression levels after tracing for 12 days. On the other hand, neonatal-TD Treg present an increased Nrp1 expression level compared with adult-TD co

关键词： T lineage tracing Treg neonatal thymus-derived T cells neonatal thymus-derived Treg single-cell RNA sequencing

Regulation of CD8^(+)T memory and exhaustion by the mTOR signals

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Cellular & Molecular Immunology 2023年第9期20卷 1023-1039页

作者： Yao Chen Ziyang Xu Hongxiang Sun Xinxing Ouyang Yuheng Han Haihui Yu ningbo wu Yiting Xie Bing Su Shanghai Institute of Immunology Department of Immunology and Microbiologyand The Ministry of Education Key Laboratory of Cell Death and DifferentiationShanghai Jiao Tong University School of MedicineShanghai 200025China Department of Tumor Biology Shanghai Chest HospitalShanghai Jiao Tong University School of MedicineShanghai 200025China Center for Immune-Related Diseases at Shanghai Institute of Immunology Department of GastroenterologyRuijin HospitalShanghai Jiao Tong University School of MedicineShanghai 200025China Shanghai Jiao Tong University School of Medicine–Yale Institute for Immune Metabolism Shanghai Jiao Tong University School of MedicineShanghaiChina Key Laboratory of Molecular Radiation Oncology of Hunan Province Xiangya HospitalCentral South UniversityChangshaChina

CD8^(+)T cells are the key executioners of the adaptive immune arm,which mediates antitumor and antiviral ***ïve CD8^(+)T cells develop in the thymus and are quickly activated in the periphery after encountering a cognate antigen,which induces these cells to proliferate and differentiate into effector cells that fight the initial ***,a fraction of these cells become long-lived memory CD8^(+)T cells that combat future ***,the generation and maintenance of memory cells is profoundly affected by various in vivo conditions,such as the mode of primary activation(e.g.,acute *** immunization)or fluctuations in host metabolic,inflammatory,or aging ***,many T cells may be lost or become exhausted and no longer *** intracellular signaling pathways,transcription factors,epigenetic modifications,and metabolic processes are involved in this ***,understanding the cellular and molecular basis for the generation and fate of memory and exhausted CD8^(+)cells is central for harnessing cellular *** this review,we focus on mammalian target of rapamycin(mTOR),particularly signaling mediated by mTOR complex(mTORC)2 in memory and exhausted CD8^(+)T cells at the molecular level.

关键词： mTOR Sin1 CD8^(+)T cell T-cell memory T-cell exhaustion

Identification of two migratory colon ILC2 populations differentially expressing IL-17A and IL-5/IL-13

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Science China(Life Sciences) 2023年第1期66卷 67-80页

作者： Hongzhi Liu Liang Li Yanyun Hao Jialu Li Zhaoyuan Liu Jingjing Qi Jingjing Zhang ningbo wu Dandan wu Caixia Gao Lei Chen Lei Shen Jinke Cheng Bing Su Shanghai Institute of Immunology Department of Immunology and Microbiologythe Ministry of Education Key Laboratory of Cell Death and DifferentiationShanghai Jiao Tong University School of MedicineShanghai 200025China Department of Biochemistry and Molecular Cell Biology Shanghai Key Laboratory for Tumor Microenvironment and InflammationShanghai Jiao Tong University School of MedicineShanghai 200025China State Key Laboratory for Oncogenes and Related Genes Key Laboratory of Gastroenterology and HepatologyMinistry of HealthDivision of Gastroenterology and HepatologyShanghai Institute of Digestive DiseaseRenji HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghai Cancer InstituteShanghai Institute of Digestive DiseaseShanghai 200001China Department of Biliary and Pancreatic Surgery Renji HospitalShanghai Jiao Tong University School of MedicineShanghai 200025China

Group 2 innate lymphoid cells(ILC2s)play important tissue resident roles in anti-parasite immunity,allergic immune response,tissue homeostasis,and tumor ***2s are considered tissue resident cells with little proliferation at steady *** studies have shown that a subset of small intestinal ILC2s could leave their residing tissues,circulate and migrate to different organs,including lung,liver,mesenteric LN and spleen,upon ***,it remains unknown whether other ILC populations with migratory behavior *** this study,we find two major colon ILC2 populations with potential to migrate to the lung in response to IL-25 *** subset expresses IL-17A and resembles inflammatory ILC2s(iILC2s)but lacks CD27 expression,whereas the other expresses CD27 but not *** addition,the IL-17A^(+)ILC2s express lower levels of CD127,CD25,and ST2 than CD27^(+)ILC2s,which express higher levels of IL-5 and ***,we found that both colon ILC2 populations still maintained their colonic features of preferential expression of IL-17A and CD27,IL-5/IL-13,***,our study identifies two migratory colon ILC2 subsets with unique surface markers and cytokine profiles which are critical in regulating lung and colon immunity and homeostasis.

关键词： colon ILC2s IL-17A CD27 iILC2 lung inflammation

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Research of the morphology of Zhejiang coastal port cities

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Journal of Geographical Sciences 1998年第1期8卷 65-71页

作者： Wang YichengDept of Geography, ningbo Normal University, ningbo 315211, CHINAwu QiyanDept of Geography, Liaoning Normal University, Dalian 116029, CHINA

Lince port cities are the first new force of China's economy to outside world, they have been developing rapidly under the open policy In this article, Zhejiang's coast port cities are taken as research subject Through intensive analysis of a great number of data which had been obtained during onthespot investigations, the regional conditions, superior and inferior conditions for development of Zhejiang's coast port cities are systematically surveyed, the development of their overall distribution trait, internal structure morphology and outer morphology is examined and the development trend of port city allocation is identified which provide theoretical basis for the further development of port cities

关键词： Zhejiang port cities port distribution pattern

Metabolic regulation of T cell development by Sinl-mTORC2 is mediated by pyruvate kinase M2

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Journal of Molecular Cell Biology 2019年第2期11卷 93-106页

作者： Xinxing Ouyang Yuheng Han Guojun Qu Man Li ningbo wu Hongzhi Liu Omotooke Arojo Hongxiang Sun Xiaobo Liu Dou Liu Lei Chen Qiang Zou Bing Su Shanghai Institute of Immunology Department of Immunology and Microbiology Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education Shanghai Jiao Tong University School of Medicine Shanghai 200025 China Department of Immunobiology and the Vascular Biology and Therapeutics Program Yale University School of Medicine 333 Cedar Street New Haven CT 06520 USA

Glucose metabolism plays a key role in thymocyte development. The mammalian target of rapamycin complex 2 (mT0RC2) is a critical regulator of cell growth and metabolism, but its role in early thymocyte development and metabolism has not been fully studied. We show here that genetic ablation of Sinl, an essential component of mTORC2, in T lineage cells results in severely impaired thymocyte development at the CD4^- CD8^- double negative (DN) stages but not at the CD4^+ CD8^+ double positive (DP) or later stages. Notably, Sinl-deficient DN thymocytes show markedly reduced proliferation and ***, we discover that the M2 isoform of pyruvate kinase (PKM2) is a novel and crucial Sinl effector in promoting DN thymocyte development and metabolism. At the molecular level, we show that Sinl-mTORC2 controls PKM2 expression through an AKT-dependent PPAR-y nuclear translocation. Together, our study unravels a novel mTORC2-PPAR-γ-PKM2 pathway in immune-metabolic regulation of early thymocyte development.

关键词： mT0RC2 Sinl thymocyte development PPAR-γ PKM2 metabolism

MAP3K2 augments Th1 cell differentiation via IL-18 to promote T cell-mediated colitis

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Science China(Life Sciences) 2021年第3期64卷 389-403页

作者： ningbo wu Dongping Chen Hongxiang Sun Jianmei Tan Yao Zhang Tianyu Zhang Yuheng Han Hongzhi Liu Xinxing Ouyang Xiao-Dong Yang Xiaoyin Niu Jie Zhong Zhengting Wang Bing Su Shanghai Institute of Immunology Department of Immunology and Microbiologyand Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of EducationShanghai JiaoTong University School of MedicineShanghai 200025China Shanghai JiaoTong University School of Medicine-Yale Institute for Immune Metabolism Shanghai JiaoTong University School of MedicineShanghai 200025China Department of Gastroenterology Ruijin HospitalShanghai JiaoTong University School of MedicineShanghai 200025China

T cell-mediated immunity in the intestine is stringently controlled to ensure proper immunity against pathogenic microbes and to prevent autoimmunity,a known cause of inflammatory bowel ***,precisely how T cells regulate intestine immunity remains to be fully *** this study,we found that mitogen-activated protein kinase kinase kinase 2(MAP3K2)is required for the CD4^(+)T cell-mediated inflammation in the *** a T cell transfer colitis model,we found that MAP3K2-deficient naïve CD4^(+)T cells had a dramatically reduced ability to induce colitis compared to wild type T *** addition,significantly fewer IFN-γ-but more IL-17A-producing CD4^(+)T cells in the intestines of mice receiving MAP3K2-deficient T cells than in those from mice receiving wild type T cells was ***,under well-defined in vitro differentiation conditions,MAP3K2-deficient naïve T cells were not impaired in their ability to differentiate into Th1,Th17 and ***,the MAP3K2-regulated colitis severity was mediated by Th1 but not Th17 cells in the *** the molecular level,we showed that MAP3K2-mediated Th1 cell differentiation in the intestine was regulated by IL-18 and required specific JNK ***,our study reveals a novel regulatory role of MAP3K2 in intestinal T cell immunity via the IL-18-MAP3K2-JNK axis and may provide a novel target for intervention in T cell-mediated colitis.

关键词： MAP3K2 IL-18 Th1 T cell immunity colitis

A NEW ASM DESIGN METHOD

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Journal of Electronics(China) 1993年第4期10卷 379-383页

作者：庄南吴浩敏 Computer Center Ningbo Normal UniversityNingbo315211 Department of Electronic Engineering Hongzhou UniversityHongzhou 310028

A novel ASM one-zero-hot design method based upon ternary flip-flops with binaryconstruction as storage cells is presented.

关键词： Flip-flops Sequential circuits Multiple-valued logic

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澳大利亚南极外大陆架划界案评析

太平洋学报 2015年第7期23卷 9-16页

作者：吴宁铂复旦大学上海200438

2004年,澳大利亚在7个南极主权要求国中率先向联合国大陆架界限委员会提交进入南纬60°以南——《南极条约》适用区域的200海里外大陆架划界申请案。1在坚持"南极主权"的前提下,澳大利亚对南极领地外大陆架与亚南极岛屿外大陆架划界问... 详细信息

2004年,澳大利亚在7个南极主权要求国中率先向联合国大陆架界限委员会提交进入南纬60°以南——《南极条约》适用区域的200海里外大陆架划界申请案。1在坚持"南极主权"的前提下,澳大利亚对南极领地外大陆架与亚南极岛屿外大陆架划界问题采取了高度灵活的区分政策,在维持其南极领土主张现状的同时,又通过对《南极条约》体系及《联合国海洋法公约》巧妙地解读与运用,确保了本国利益的最大化。不过,由此引发的相关国际法问题还有待进一步剖析和解决。

关键词：外大陆架《南极条约》亚南极岛屿大陆架界限委员会《联合国海洋法公约》