In this paper, we report a multiple sequence alignment result on the basis of 10 amino acid sequences of the M protein, which come from different coronaviruses (4 SARS associated and 6 others known). The alignment mo...In this paper, we report a multiple sequence alignment result on the basis of 10 amino acid sequences of the M protein, which come from different coronaviruses (4 SARS associated and 6 others known). The alignment model was based on the profile HMM (Hidden Markov Model), and the model training was implemented through the SAHMM (Self Adapting Hidden Markov Model) software developed by the authors.展开更多
目的:探讨不同浓度重组Canstatin蛋白对碱烧伤后小鼠角膜基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)及其组织抑制剂-2(tissue inhibitor of metalloproteinase-2,TIMP-2)表达的影响及其调节作用。方法:BALB/c小鼠60只随机分...目的:探讨不同浓度重组Canstatin蛋白对碱烧伤后小鼠角膜基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)及其组织抑制剂-2(tissue inhibitor of metalloproteinase-2,TIMP-2)表达的影响及其调节作用。方法:BALB/c小鼠60只随机分为实验组A、实验B及对照组C,每组20只。采用1mol/L氢氧化钠溶液烧伤小鼠右眼角膜,建立炎症性角膜碱烧伤动物模型,分别予以A组、B组重组Canstatin蛋白3μg/mL、5μg/mL点右眼,4次/d,对照组C组予以生理眼水点右眼。在碱烧伤后第1、3、7、14d以形态学分析评价角膜上皮损伤面积及新生血管生长的情况,并于碱烧伤后第1、3、7、14d应用Western-blot检测角膜MMP-2和TIMP-2的表达,增强化学发光法(ECL)对结果进行分析。结果:形态学分析显示,A组和B组小鼠在碱烧伤后第3d起各时间点角膜上皮缺损面积均小于对照组(P<0.01),角膜新生血管均得到抑制,CNV面积明显小于对照组(P<0.01)。Western-blot结果显示,碱烧伤后各时间点MMP-2的表达,A组和B组均明显低于对照组(P<0.01),TIMP-2的表达高于对照组(P<0.01),且A组和B组间MMP-2的表达在第14d比较差异有统计学意义(P<0.05),TIMP-2的表达在第7d及第14d比较差异有统计学意义(P<0.05)。结论:重组Canstatin蛋白可通过抑制角膜细胞及浸润的炎性细胞产生MMP-2,促进TIMP-2表达,从而抑制和延迟碱烧伤后角膜融解的发生和发展,对碱烧伤后角膜的重塑起着重要作用。展开更多
The matrix protein 2 (M2) is a preferred target for developing a universal vaccine against the influenza A virus (IAV). This study aimed to develop a method for assessing antibody-dependent cell-mediated cytotoxicity ...The matrix protein 2 (M2) is a preferred target for developing a universal vaccine against the influenza A virus (IAV). This study aimed to develop a method for assessing antibody-dependent cell-mediated cytotoxicity (ADCC) associated with M2-based immunization in mice. We first established a stable cell line derived from mouse lymphoma cells (YAC-1) expressing M2 of H3N2. This cell line, designated as YAC-1-M2, was generated using a second-generation lentiviral tricistronic plasmid system to transduce the M2 gene into YAC-1 cells. The ADCC effect induced by polyclonal antibodies targeting matrix protein 2 ectodomain (M2e) was demonstrated by YAC-1-M2 cell lysis by natural killer cells (NK) derived from mice, in the presence of anti-M2 antibodies obtained from mice immunized with an mRNA vaccine based on M2e. This ADCC effect was found to be stronger compared to the effect induced by monoclonal antibodies (14C2) against M2. Moreover, the ADCC effect was enhanced as the effector-to-target ratio of NK to YAC-1-M2 cells increased. In conclusion, we established a novel method to detect ADCC of M2 of IAV, which paves the way for the development of an M2-based universal vaccine against IAV and an in-depth analysis of its mechanism of broad-spectrum immune protection in mice.展开更多
文摘In this paper, we report a multiple sequence alignment result on the basis of 10 amino acid sequences of the M protein, which come from different coronaviruses (4 SARS associated and 6 others known). The alignment model was based on the profile HMM (Hidden Markov Model), and the model training was implemented through the SAHMM (Self Adapting Hidden Markov Model) software developed by the authors.
文摘目的:探讨不同浓度重组Canstatin蛋白对碱烧伤后小鼠角膜基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)及其组织抑制剂-2(tissue inhibitor of metalloproteinase-2,TIMP-2)表达的影响及其调节作用。方法:BALB/c小鼠60只随机分为实验组A、实验B及对照组C,每组20只。采用1mol/L氢氧化钠溶液烧伤小鼠右眼角膜,建立炎症性角膜碱烧伤动物模型,分别予以A组、B组重组Canstatin蛋白3μg/mL、5μg/mL点右眼,4次/d,对照组C组予以生理眼水点右眼。在碱烧伤后第1、3、7、14d以形态学分析评价角膜上皮损伤面积及新生血管生长的情况,并于碱烧伤后第1、3、7、14d应用Western-blot检测角膜MMP-2和TIMP-2的表达,增强化学发光法(ECL)对结果进行分析。结果:形态学分析显示,A组和B组小鼠在碱烧伤后第3d起各时间点角膜上皮缺损面积均小于对照组(P<0.01),角膜新生血管均得到抑制,CNV面积明显小于对照组(P<0.01)。Western-blot结果显示,碱烧伤后各时间点MMP-2的表达,A组和B组均明显低于对照组(P<0.01),TIMP-2的表达高于对照组(P<0.01),且A组和B组间MMP-2的表达在第14d比较差异有统计学意义(P<0.05),TIMP-2的表达在第7d及第14d比较差异有统计学意义(P<0.05)。结论:重组Canstatin蛋白可通过抑制角膜细胞及浸润的炎性细胞产生MMP-2,促进TIMP-2表达,从而抑制和延迟碱烧伤后角膜融解的发生和发展,对碱烧伤后角膜的重塑起着重要作用。
基金the National Key Research and Development Program of China(2021YFC2300101).
文摘The matrix protein 2 (M2) is a preferred target for developing a universal vaccine against the influenza A virus (IAV). This study aimed to develop a method for assessing antibody-dependent cell-mediated cytotoxicity (ADCC) associated with M2-based immunization in mice. We first established a stable cell line derived from mouse lymphoma cells (YAC-1) expressing M2 of H3N2. This cell line, designated as YAC-1-M2, was generated using a second-generation lentiviral tricistronic plasmid system to transduce the M2 gene into YAC-1 cells. The ADCC effect induced by polyclonal antibodies targeting matrix protein 2 ectodomain (M2e) was demonstrated by YAC-1-M2 cell lysis by natural killer cells (NK) derived from mice, in the presence of anti-M2 antibodies obtained from mice immunized with an mRNA vaccine based on M2e. This ADCC effect was found to be stronger compared to the effect induced by monoclonal antibodies (14C2) against M2. Moreover, the ADCC effect was enhanced as the effector-to-target ratio of NK to YAC-1-M2 cells increased. In conclusion, we established a novel method to detect ADCC of M2 of IAV, which paves the way for the development of an M2-based universal vaccine against IAV and an in-depth analysis of its mechanism of broad-spectrum immune protection in mice.
