Background: The chemokine eotaxin-2 is a potent chemoattractant for inflammatory cells, the predominants of which are eosinophils. Human and murine atherosclerotic plaques are known to exhibit inflammatory phenotypes ...Background: The chemokine eotaxin-2 is a potent chemoattractant for inflammatory cells, the predominants of which are eosinophils. Human and murine atherosclerotic plaques are known to exhibit inflammatory phenotypes where a complex interaction of cytokine and chemokines plays a role. We tested the hypothesis that eotaxin-2 (eo-2) plays a causative role in the initiation and progression of experimental atherosclerosis. Methods and Results: Sera collected from atherosclerotic ApoE knockout (KO) mice, exhibited significantly higher levels of eo-2 compared to sera collected from their background age matched C57BL/6 litters by ELISA. Moreover, levels of eo-2 were higher in old atherosclerotic ApoE KO mice than in young animals. Similarly, the expression level of the eo-2 receptor, CCR3, was increased in splenocytes of old ApoE compared to the young littermates. Administration of polyclonal blocking antibodies to eotaxin-2 resulted in a significant reduction of early atherosclerotic plaques in ApoE KO mice whereas prolonged treatment of mice with advanced plaques led to atheroma stabilization. A monoclonal antibody (D8) prepared against eo-2 attenuated adhesion of lymphocytes to fibronectin and potently inhibited their migration towards VEGF. Monoclonal blocking antibodies to eo-2 also significantly reduced atherosclerotic plaques in ApoE KO mice. Conclusion: Eo-2 serum levels are elevated in sera of ApoE KO mice with experimental atherosclerosis and its blockade is associated with reduced fatty streak accumulation and increased plaque stabilization.展开更多
AIM: To study the effect of blocking the eo-2 pathwayon the development and severity of experimental autoimmune encephalomyelitis(EAE). METHODS: We produced m Ab directed against eo-2,named D8. MOG35-55 induced-EAE mi...AIM: To study the effect of blocking the eo-2 pathwayon the development and severity of experimental autoimmune encephalomyelitis(EAE). METHODS: We produced m Ab directed against eo-2,named D8. MOG35-55 induced-EAE mice were dailyintravenously injected with either 25 μg or 100 μg D8,or with vehicle control alone [phosphate-buffered saline(PBS)], starting from day 0 post immunization and weremonitored for EAE clinical score(n = 10 in each group).Mice were sacrificed on day 58 and their sera were assessed for the presence of anti-myelin oligodendrocyteglycoprotein(anti-MOG) antibodies autoantibodies, aswell as for the profile of pro-inflammatory cytokines andchemokines. Histological analysis of brain sections wasperformed by hematoxylin and eosin staining.RESULTS: Daily treatment of EAE induced mice with D8 significantly decreased the severity of EAE symptoms. Treatment with both concentrations of D8 ameliorated EAE symptoms compared to PBS treated mice, starting from day 42 post immunization(0.89 ± 0.35 in D8 25 μg and D8 100 μg treated groups vs 2.11 ± 0.38 in the PBS treated group, P = 0.03). A significant improvement in EAE clinical score compared to total Ig G treated mice was observed with the higher concentration of D8(0.81 ± 0.38 in D8 100 μg treated group vs 2.11 ± 0.31 in Ig G1 treated group, on day 56 post immunization, P = 0.04). D8 treated mice with EAE did not significantly exhibit lower sera levels of anti-MOG autoantibodies compared to Ig G-treated mice. However, they expressed lower sera levels of the pro-inflammatory cytokines: tumor necrosis factor(7.8 ± 0.2 pg/m L in D8 100 μg treated mice vs 19.9 ± 3.4 pg/m L in Ig G treated mice, P = 0.005) and interferon-gamma(1.4 ± 0.6 pg/m L in D8 100 μg treated mice vs 3.6 ± 0.4 pg/m L in Ig G treated mice, P = 0.02), as well as reduced levels of the chemokine macrophage chemoattractant protein-1(27.2 ± 3.1 pg/m L in D8 100 μg treated mice vs 63.7 ± 12.3 pg/m L in Ig G treated mice, P = 0.03). These findings indicate that blocking the eo-2 pathway in EAE may affect not only eosinophil infiltration into the central nervous system(CNS), but also have an effect on monocytes and T cells, but not humoral, mediated responses. Histological analysis of the brains of D8 treated mice with EAE support that this treatment decreases immune cells infiltrates in the CNS.CONCLUSION: Taken together, these findings suggest a role for eo-2 in EAE pathogenesis and consequentially may support a therapeutic potential of anti-eo-2 neutralizing m Ab in multiple sclerosis.展开更多
Objective To study the role of eotaxin and eotaxin-2 expression by Th2 cytokine and analyze their relationship in normal human bronchial epithelial cell line-BEAS-2B cell. Methods Levels of eotaxin mRNA and protein ex...Objective To study the role of eotaxin and eotaxin-2 expression by Th2 cytokine and analyze their relationship in normal human bronchial epithelial cell line-BEAS-2B cell. Methods Levels of eotaxin mRNA and protein expression in the bronchial epithelial cell line BEAS-2B cell were determined with RT-PCR and ELISA. We also used RT-PCR to evaluate eotaxin-2 expression under the regulation of Th2 cytokine IL-4 and IL-13 as well as proinflammatory agent-TNFα. Results Eotaxin mRNA expression was the highest at the time point of 12h under the stimulation of TNF-α. While Th2 cytokine IL-4 and IL-13 had the amplification effect on the expression. Eotaxin protein was also elevated with the combination stimulation of proinflammatory agent TNF-α and IL-4 in dose and time dependent manner(P< 0.01). These results were also seen when the cells were stimulated by TNF-α and IL-13. Eotaxin-2 mRNA expression was the highest at the time point of 8h. The expression evaluated by semi-quantitative RT-PCR also elevated under the co-stimulation of TNF-α and IL-4 or TNF-α and IL-13 and it should significantly correlate with Eotaxin(P<0.05). Conclusion This study demonstrated that Th2 cytokine like IL-4 and IL-13 enhances eotaxin and eotaxin-2 expression when co-stimulated with proinflammatory agent TNF-α. These results showed that Th2 cytokines existence is the strong evidence for bronchial epithelial cells taking part in the allergic inflammation especially in eosinophils recruitment.展开更多
目的探讨转移相关蛋白2(MTA2)对前列腺癌预后的预测价值及在癌转移过程中的作用。方法收集空军军医大学第一附属医院2018年1月~2020年6月收治的50例前列腺癌患者的癌组织及配对癌旁组织样本用于免疫组织化学分析。根据MTA2蛋白的表达水...目的探讨转移相关蛋白2(MTA2)对前列腺癌预后的预测价值及在癌转移过程中的作用。方法收集空军军医大学第一附属医院2018年1月~2020年6月收治的50例前列腺癌患者的癌组织及配对癌旁组织样本用于免疫组织化学分析。根据MTA2蛋白的表达水平将样本分为两组,染色评分≥4为高表达组,<4为低表达组。对人前列腺癌细胞系(PC-3)转染shMTA2(MTA2-shRNA-pLKO.1)来沉默MTA2作为shMTA2组,转染Luc-shRNA-pLKO.1的细胞作为阴性对照组(shNC组),未转染的细胞作为空白对照组(control组)。通过MTT法检测细胞活力,Transwells实验进行体外迁移和侵袭分析。通过qRT-PCR或Western Blot检测细胞中MTA2、Eotaxin-1、CCR3、p-ERK1/2、t-ERK1/2和MMP-3的表达。结果前列腺癌组织中MTA2的染色评分显著高于癌旁组织(5.16±0.87 vs 2.34±0.39,t=9.221,P<0.001)。低表达组患者中出现淋巴结转移率为14.29%(2/14),高表达组患者中出现淋巴结转移率为52.78%(19/36),两组比较差异有统计学意义(χ^(2)=6.131,P=0.013)。与低表达组相比,高表达组患者的生存时间更长(χ^(2)=4.756,P=0.029)。与空白对照组相比,shMTA2组的细胞活力降低了56.87%(P<0.001),细胞迁移数量降低了65.80%(P<0.001),细胞侵袭数量降低了56.35%(P<0.001),Eotaxin-1、CCR3、p-ERK1/2和MMP-3的蛋白相对表达量依次降低了76.03%、69.12%、72.32%和54.67%(P<0.001)。结论前列腺癌组织中MTA2的表达水平升高且与患者的预后有关,沉默MTA2可降低前列腺癌细胞的迁移和侵袭能力,并抑制Eotaxin-CCR3-ERK1/2-MMP-3轴。展开更多
文摘Background: The chemokine eotaxin-2 is a potent chemoattractant for inflammatory cells, the predominants of which are eosinophils. Human and murine atherosclerotic plaques are known to exhibit inflammatory phenotypes where a complex interaction of cytokine and chemokines plays a role. We tested the hypothesis that eotaxin-2 (eo-2) plays a causative role in the initiation and progression of experimental atherosclerosis. Methods and Results: Sera collected from atherosclerotic ApoE knockout (KO) mice, exhibited significantly higher levels of eo-2 compared to sera collected from their background age matched C57BL/6 litters by ELISA. Moreover, levels of eo-2 were higher in old atherosclerotic ApoE KO mice than in young animals. Similarly, the expression level of the eo-2 receptor, CCR3, was increased in splenocytes of old ApoE compared to the young littermates. Administration of polyclonal blocking antibodies to eotaxin-2 resulted in a significant reduction of early atherosclerotic plaques in ApoE KO mice whereas prolonged treatment of mice with advanced plaques led to atheroma stabilization. A monoclonal antibody (D8) prepared against eo-2 attenuated adhesion of lymphocytes to fibronectin and potently inhibited their migration towards VEGF. Monoclonal blocking antibodies to eo-2 also significantly reduced atherosclerotic plaques in ApoE KO mice. Conclusion: Eo-2 serum levels are elevated in sera of ApoE KO mice with experimental atherosclerosis and its blockade is associated with reduced fatty streak accumulation and increased plaque stabilization.
文摘AIM: To study the effect of blocking the eo-2 pathwayon the development and severity of experimental autoimmune encephalomyelitis(EAE). METHODS: We produced m Ab directed against eo-2,named D8. MOG35-55 induced-EAE mice were dailyintravenously injected with either 25 μg or 100 μg D8,or with vehicle control alone [phosphate-buffered saline(PBS)], starting from day 0 post immunization and weremonitored for EAE clinical score(n = 10 in each group).Mice were sacrificed on day 58 and their sera were assessed for the presence of anti-myelin oligodendrocyteglycoprotein(anti-MOG) antibodies autoantibodies, aswell as for the profile of pro-inflammatory cytokines andchemokines. Histological analysis of brain sections wasperformed by hematoxylin and eosin staining.RESULTS: Daily treatment of EAE induced mice with D8 significantly decreased the severity of EAE symptoms. Treatment with both concentrations of D8 ameliorated EAE symptoms compared to PBS treated mice, starting from day 42 post immunization(0.89 ± 0.35 in D8 25 μg and D8 100 μg treated groups vs 2.11 ± 0.38 in the PBS treated group, P = 0.03). A significant improvement in EAE clinical score compared to total Ig G treated mice was observed with the higher concentration of D8(0.81 ± 0.38 in D8 100 μg treated group vs 2.11 ± 0.31 in Ig G1 treated group, on day 56 post immunization, P = 0.04). D8 treated mice with EAE did not significantly exhibit lower sera levels of anti-MOG autoantibodies compared to Ig G-treated mice. However, they expressed lower sera levels of the pro-inflammatory cytokines: tumor necrosis factor(7.8 ± 0.2 pg/m L in D8 100 μg treated mice vs 19.9 ± 3.4 pg/m L in Ig G treated mice, P = 0.005) and interferon-gamma(1.4 ± 0.6 pg/m L in D8 100 μg treated mice vs 3.6 ± 0.4 pg/m L in Ig G treated mice, P = 0.02), as well as reduced levels of the chemokine macrophage chemoattractant protein-1(27.2 ± 3.1 pg/m L in D8 100 μg treated mice vs 63.7 ± 12.3 pg/m L in Ig G treated mice, P = 0.03). These findings indicate that blocking the eo-2 pathway in EAE may affect not only eosinophil infiltration into the central nervous system(CNS), but also have an effect on monocytes and T cells, but not humoral, mediated responses. Histological analysis of the brains of D8 treated mice with EAE support that this treatment decreases immune cells infiltrates in the CNS.CONCLUSION: Taken together, these findings suggest a role for eo-2 in EAE pathogenesis and consequentially may support a therapeutic potential of anti-eo-2 neutralizing m Ab in multiple sclerosis.
文摘Objective To study the role of eotaxin and eotaxin-2 expression by Th2 cytokine and analyze their relationship in normal human bronchial epithelial cell line-BEAS-2B cell. Methods Levels of eotaxin mRNA and protein expression in the bronchial epithelial cell line BEAS-2B cell were determined with RT-PCR and ELISA. We also used RT-PCR to evaluate eotaxin-2 expression under the regulation of Th2 cytokine IL-4 and IL-13 as well as proinflammatory agent-TNFα. Results Eotaxin mRNA expression was the highest at the time point of 12h under the stimulation of TNF-α. While Th2 cytokine IL-4 and IL-13 had the amplification effect on the expression. Eotaxin protein was also elevated with the combination stimulation of proinflammatory agent TNF-α and IL-4 in dose and time dependent manner(P< 0.01). These results were also seen when the cells were stimulated by TNF-α and IL-13. Eotaxin-2 mRNA expression was the highest at the time point of 8h. The expression evaluated by semi-quantitative RT-PCR also elevated under the co-stimulation of TNF-α and IL-4 or TNF-α and IL-13 and it should significantly correlate with Eotaxin(P<0.05). Conclusion This study demonstrated that Th2 cytokine like IL-4 and IL-13 enhances eotaxin and eotaxin-2 expression when co-stimulated with proinflammatory agent TNF-α. These results showed that Th2 cytokines existence is the strong evidence for bronchial epithelial cells taking part in the allergic inflammation especially in eosinophils recruitment.
文摘目的探讨转移相关蛋白2(MTA2)对前列腺癌预后的预测价值及在癌转移过程中的作用。方法收集空军军医大学第一附属医院2018年1月~2020年6月收治的50例前列腺癌患者的癌组织及配对癌旁组织样本用于免疫组织化学分析。根据MTA2蛋白的表达水平将样本分为两组,染色评分≥4为高表达组,<4为低表达组。对人前列腺癌细胞系(PC-3)转染shMTA2(MTA2-shRNA-pLKO.1)来沉默MTA2作为shMTA2组,转染Luc-shRNA-pLKO.1的细胞作为阴性对照组(shNC组),未转染的细胞作为空白对照组(control组)。通过MTT法检测细胞活力,Transwells实验进行体外迁移和侵袭分析。通过qRT-PCR或Western Blot检测细胞中MTA2、Eotaxin-1、CCR3、p-ERK1/2、t-ERK1/2和MMP-3的表达。结果前列腺癌组织中MTA2的染色评分显著高于癌旁组织(5.16±0.87 vs 2.34±0.39,t=9.221,P<0.001)。低表达组患者中出现淋巴结转移率为14.29%(2/14),高表达组患者中出现淋巴结转移率为52.78%(19/36),两组比较差异有统计学意义(χ^(2)=6.131,P=0.013)。与低表达组相比,高表达组患者的生存时间更长(χ^(2)=4.756,P=0.029)。与空白对照组相比,shMTA2组的细胞活力降低了56.87%(P<0.001),细胞迁移数量降低了65.80%(P<0.001),细胞侵袭数量降低了56.35%(P<0.001),Eotaxin-1、CCR3、p-ERK1/2和MMP-3的蛋白相对表达量依次降低了76.03%、69.12%、72.32%和54.67%(P<0.001)。结论前列腺癌组织中MTA2的表达水平升高且与患者的预后有关,沉默MTA2可降低前列腺癌细胞的迁移和侵袭能力,并抑制Eotaxin-CCR3-ERK1/2-MMP-3轴。
