Background:Microglia and brain macrophages contribute significantly to the tumor microenvironment in highly malignant glioblastoma where they are considered important drivers of tumor progression.A better understandin...Background:Microglia and brain macrophages contribute significantly to the tumor microenvironment in highly malignant glioblastoma where they are considered important drivers of tumor progression.A better understanding of the role of the brain macrophages present in glioblastoma appears crucial for improving therapeutic outcomes,especially in the context of novel immunotherapeutic approaches.Methods:We investigated the regulation of two well-established markers for microglia and brain macrophages,IBA1 and CD163,in relation to glioblastoma tumor necrosis using immunohistochemistry and modality fusion heatmaps of whole slide images obtained from adjacent tissue sections.Results:IBA1 and CD163 showed remarkable differences in relation to glioblastoma tumor necrosis.Generally,IBA1 immunoreactive cells were far less common in necrotic tissue areas than CD163-expressing cells.We also found extensive and frequently diffuse extracellular CD163 deposition,especially in hypocellular necrobiotic tumor regions where IBA1 was typically absent.Conclusions:Resident microglia seem more likely to be important for the diffuse infiltration of glioma cells in hypercellular tissue areas,whereas myeloid macrophages may be the main macrophage population in the wake of tumor necrosis.Since the necrotic niche with its interactions between microglia,brain macrophages,and glioblastoma/glioma stem cells is increasingly recognised as an important factor in tumor progression,further detailed studies of the macrophage populations in glioblastoma are warranted.展开更多
Innate immunity,particularly macrophages,is critical for intestinal homeostasis.Sulforaphane,a dietary isothiocyanate from cruciferous vegetables,has been reported to protect against intestinal inflammation.However,th...Innate immunity,particularly macrophages,is critical for intestinal homeostasis.Sulforaphane,a dietary isothiocyanate from cruciferous vegetables,has been reported to protect against intestinal inflammation.However,the role of macrophages in sulforaphane mediated intestinal inflammation and the underlying molecular mechanisms have not been studied yet.In this study,sulforaphane effectively attenuated dextran sodium sulphate(DSS)induced intestinal inflammation in murine model.Of note,sulforaphane skewed the switching from classically(M1)to alternatively(M2)activated phenotype both in intestinal and bone marrow-derived macrophages(BMDMs).The expression levels of M1 associated maker genes induced by DSS or lipopolysaccharide(LPS)plus interferon gamma-γ(IFN-γ)were suppressed by sulforaphane while M2 marker gene expression levels were improved.This resulted in alteration of inflammatory mediators,particularly interleukin-10(IL-10),both in colon tissues and culture medium of BMDMs.Subsequently,IL-10 was found to mediate the sulforaphane induced M2 phenotype switching of BMDMs through the activation of STAT3 signaling.This was confirmed by immunofluorescence analysis with increased number of p-STAT3-positive cells in the colon sections.Moreover,anti-IL-10 neutralizing antibody significantly interfered M2 phenotyping of BMDMs induced by sulforaphane with reduced STAT3 phosphorylation.Findings here introduced a potential utilization of sulforaphane for intestinal inflammation treatment with macrophages as the therapeutic targets.展开更多
目的探讨炎症条件的动物输注贮存红细胞对巨噬细胞(BMDMs)的调节作用以及贮存红细胞输注与细菌感染引发炎症反应的关系。方法将6~8周龄成年雄性C57BL/6小鼠[(18~22)g/只]40只随机均分为实验组和实验对照组(对照组),均通过动物尾静脉注...目的探讨炎症条件的动物输注贮存红细胞对巨噬细胞(BMDMs)的调节作用以及贮存红细胞输注与细菌感染引发炎症反应的关系。方法将6~8周龄成年雄性C57BL/6小鼠[(18~22)g/只]40只随机均分为实验组和实验对照组(对照组),均通过动物尾静脉注射铜绿假单胞菌200μL/只,并使用吸入式麻醉剂异氟烷(1%~3%)麻醉后,通过小鼠眼后静脉丛,实验组输注鼠源贮存悬浮红细胞(>14 d)400μL/只、对照组每只输注等量新鲜悬浮红细胞(贮存<24 h);于输注后2、4、8 h脱就猝死各结束2组小鼠生命5只,摘取鼠肝,体外培养铜绿假单胞菌感染(200μL/只)小鼠的股骨、胫骨骨髓来源的BMDMs,流式细胞术检测BMDMs中分化簇86(CD86)、分化簇197(CD197)[巨噬细胞1型(M1)基因特异性标志物]、分化簇209(CD209)[巨噬细胞2型(M2)基因特异性标记]表达水平,实时荧光定量PCR(qRT-PCR)法检测小鼠肝脏F4/80、M1、M2基因表达水平,并使用SPSS17.0统计学软件分析数据。结果实验组与对照组BMDMs中CD86和CD197的表达(%)分别为8688±1.01 vs 79.24±2.65、38.59±3.73 vs 25.95±0.86(P<0.05),CD209(%)为23.88±2.23 vs 21.91±3.58(P>0.05)。输注红细胞后2、4 h,小鼠肝F4/80基因表达水平实验组和对照组分别为1.83±0.11 vs 0.75±0.06、0.46±0.06 vs 0.33±0.06(P<0.05),8 h后分别为0.33±0.03 vs 0.35±0.05(P>0.05);输注红细胞2、4、8 h,小鼠肝M1基因中诱导型一氧化氮合酶(iNOS)基因表达水平实验组和对照组分别为3.44±0.20 vs 2.46±0.08、9.25±0.55 vs 2.67±0.12、2.80±0.08 vs 2.39±0.01,肿瘤坏死因子-α(TNF-α)分别为1.69±0.22 vs 1.13±0.03、1.44±0.24 vs 0.96±0.09、1.31±0.05 vs 0.96±0.06,单核细胞趋化蛋白1(MCP1)分别为4.96±0.08 vs 4.28±0.27、4.63±0.04 vs 2.07±0.09、2.28±0.19 vs 1.33±0.03(P<0.05);M2基因中精氨酸1(Arg1)基因表达水平实验组和对照组分别为0.81±0.21 vs 0.82±0.18、0.66±0.11 vs 0.58±0.09、0.39±0.17 vs 0.37±0.15,甘露糖受体C型2(Mrc2)分别为0.99±0.91 vs 0.97±0.08、0.98±0.12 vs 1.02±0.11、0.59±0.19 vs 0.57±0.08,重组蛋白163(CD163)分别为1.75±0.20 vs 1.69±0.18、0.22±0.02 vs 0.21±0.01、0.04±0.01 vs 0.03±0.01(P>0.05)。结论实验小鼠输注贮存红细胞明显促进其肝脏组织巨噬细胞朝向M1表型的极化。展开更多
基金funding but aspects of this work(computer analysis)were supported by an Australian Research Council(ARC)grant,DP150104472(MBG)The Australian Genomics and Outcomes of Glioma project was funded by a Cancer Council NSW Strategic Research Partnership Grant.
文摘Background:Microglia and brain macrophages contribute significantly to the tumor microenvironment in highly malignant glioblastoma where they are considered important drivers of tumor progression.A better understanding of the role of the brain macrophages present in glioblastoma appears crucial for improving therapeutic outcomes,especially in the context of novel immunotherapeutic approaches.Methods:We investigated the regulation of two well-established markers for microglia and brain macrophages,IBA1 and CD163,in relation to glioblastoma tumor necrosis using immunohistochemistry and modality fusion heatmaps of whole slide images obtained from adjacent tissue sections.Results:IBA1 and CD163 showed remarkable differences in relation to glioblastoma tumor necrosis.Generally,IBA1 immunoreactive cells were far less common in necrotic tissue areas than CD163-expressing cells.We also found extensive and frequently diffuse extracellular CD163 deposition,especially in hypocellular necrobiotic tumor regions where IBA1 was typically absent.Conclusions:Resident microglia seem more likely to be important for the diffuse infiltration of glioma cells in hypercellular tissue areas,whereas myeloid macrophages may be the main macrophage population in the wake of tumor necrosis.Since the necrotic niche with its interactions between microglia,brain macrophages,and glioblastoma/glioma stem cells is increasingly recognised as an important factor in tumor progression,further detailed studies of the macrophage populations in glioblastoma are warranted.
基金the financial support provided by“Modern Food Processing,Food Storage,Transportation Technology,and Equipment”State Key Research and Development Plan(2017YFD0400204)the National Science Foundation of China(31972091).
文摘Innate immunity,particularly macrophages,is critical for intestinal homeostasis.Sulforaphane,a dietary isothiocyanate from cruciferous vegetables,has been reported to protect against intestinal inflammation.However,the role of macrophages in sulforaphane mediated intestinal inflammation and the underlying molecular mechanisms have not been studied yet.In this study,sulforaphane effectively attenuated dextran sodium sulphate(DSS)induced intestinal inflammation in murine model.Of note,sulforaphane skewed the switching from classically(M1)to alternatively(M2)activated phenotype both in intestinal and bone marrow-derived macrophages(BMDMs).The expression levels of M1 associated maker genes induced by DSS or lipopolysaccharide(LPS)plus interferon gamma-γ(IFN-γ)were suppressed by sulforaphane while M2 marker gene expression levels were improved.This resulted in alteration of inflammatory mediators,particularly interleukin-10(IL-10),both in colon tissues and culture medium of BMDMs.Subsequently,IL-10 was found to mediate the sulforaphane induced M2 phenotype switching of BMDMs through the activation of STAT3 signaling.This was confirmed by immunofluorescence analysis with increased number of p-STAT3-positive cells in the colon sections.Moreover,anti-IL-10 neutralizing antibody significantly interfered M2 phenotyping of BMDMs induced by sulforaphane with reduced STAT3 phosphorylation.Findings here introduced a potential utilization of sulforaphane for intestinal inflammation treatment with macrophages as the therapeutic targets.
文摘目的探讨炎症条件的动物输注贮存红细胞对巨噬细胞(BMDMs)的调节作用以及贮存红细胞输注与细菌感染引发炎症反应的关系。方法将6~8周龄成年雄性C57BL/6小鼠[(18~22)g/只]40只随机均分为实验组和实验对照组(对照组),均通过动物尾静脉注射铜绿假单胞菌200μL/只,并使用吸入式麻醉剂异氟烷(1%~3%)麻醉后,通过小鼠眼后静脉丛,实验组输注鼠源贮存悬浮红细胞(>14 d)400μL/只、对照组每只输注等量新鲜悬浮红细胞(贮存<24 h);于输注后2、4、8 h脱就猝死各结束2组小鼠生命5只,摘取鼠肝,体外培养铜绿假单胞菌感染(200μL/只)小鼠的股骨、胫骨骨髓来源的BMDMs,流式细胞术检测BMDMs中分化簇86(CD86)、分化簇197(CD197)[巨噬细胞1型(M1)基因特异性标志物]、分化簇209(CD209)[巨噬细胞2型(M2)基因特异性标记]表达水平,实时荧光定量PCR(qRT-PCR)法检测小鼠肝脏F4/80、M1、M2基因表达水平,并使用SPSS17.0统计学软件分析数据。结果实验组与对照组BMDMs中CD86和CD197的表达(%)分别为8688±1.01 vs 79.24±2.65、38.59±3.73 vs 25.95±0.86(P<0.05),CD209(%)为23.88±2.23 vs 21.91±3.58(P>0.05)。输注红细胞后2、4 h,小鼠肝F4/80基因表达水平实验组和对照组分别为1.83±0.11 vs 0.75±0.06、0.46±0.06 vs 0.33±0.06(P<0.05),8 h后分别为0.33±0.03 vs 0.35±0.05(P>0.05);输注红细胞2、4、8 h,小鼠肝M1基因中诱导型一氧化氮合酶(iNOS)基因表达水平实验组和对照组分别为3.44±0.20 vs 2.46±0.08、9.25±0.55 vs 2.67±0.12、2.80±0.08 vs 2.39±0.01,肿瘤坏死因子-α(TNF-α)分别为1.69±0.22 vs 1.13±0.03、1.44±0.24 vs 0.96±0.09、1.31±0.05 vs 0.96±0.06,单核细胞趋化蛋白1(MCP1)分别为4.96±0.08 vs 4.28±0.27、4.63±0.04 vs 2.07±0.09、2.28±0.19 vs 1.33±0.03(P<0.05);M2基因中精氨酸1(Arg1)基因表达水平实验组和对照组分别为0.81±0.21 vs 0.82±0.18、0.66±0.11 vs 0.58±0.09、0.39±0.17 vs 0.37±0.15,甘露糖受体C型2(Mrc2)分别为0.99±0.91 vs 0.97±0.08、0.98±0.12 vs 1.02±0.11、0.59±0.19 vs 0.57±0.08,重组蛋白163(CD163)分别为1.75±0.20 vs 1.69±0.18、0.22±0.02 vs 0.21±0.01、0.04±0.01 vs 0.03±0.01(P>0.05)。结论实验小鼠输注贮存红细胞明显促进其肝脏组织巨噬细胞朝向M1表型的极化。
