Membrane-associated RING-CH-1 (MARCH1) is an E3 ubiquitin ligase which targets MHC-II, CD86 and various other molecules for degradation. It is one of the most efficient post-translational regulators of antigen present...Membrane-associated RING-CH-1 (MARCH1) is an E3 ubiquitin ligase which targets MHC-II, CD86 and various other molecules for degradation. It is one of the most efficient post-translational regulators of antigen presentation. MARCH1 is expressed in resting immature dendritic cells and B cells but can be induced in other cell types. While activation of most immune cells results in a reduction in MARCH1 gene expression, its anti-inflammatory properties are highlighted by its induction in response to IL-10. Here, we review what is known about the regulation of MARCH1 gene expression in response to IL-10 by various immune cells. We speculate on the role of MARCH1 ininfection, its differential expression pattern and the promise that this E3 ubiquitin ligase holds to influence immune responses and mitigate immune pathologies.展开更多
为进一步丰富鱼类MHC class Ⅱ基因的研究,同时也为进一步探讨低磷饲料中添加维生素D3对鱼类免疫功能可能的影响,实验利用RACE(Rapid-amplification of c DNA ends)即c DNA末端快速扩增技术,成功克隆出黄颡鱼(Pelteobagrus fulvidraco)...为进一步丰富鱼类MHC class Ⅱ基因的研究,同时也为进一步探讨低磷饲料中添加维生素D3对鱼类免疫功能可能的影响,实验利用RACE(Rapid-amplification of c DNA ends)即c DNA末端快速扩增技术,成功克隆出黄颡鱼(Pelteobagrus fulvidraco)主要组织相容性复合体(Major histocompatibility complex,MHC)class Ⅱ抗原基因,全长1074 bp,其中ORF(Open reading frame)708 bp,编码236个氨基酸,5′UTR(5′端非翻译区)78 bp,3′UTR(3′端非翻译区)259 bp。进行氨基酸序列比对分析得到:黄颡鱼MHC class Ⅱ基因ORF氨基酸序列与长吻逘(Leiocassis longirostris)的氨基酸序列相似度最高为69.5%,与锦鲤(Cyprinus carpio)的氨基酸序列相似度最低为50.4%。利用q PCR对黄颡鱼MHC class Ⅱ基因进行组织表达分析,结果表明MHC class Ⅱ在小肠、肝脏、鳃中表达较高;在肌肉、鳍条中表达较低;而在肾、脾脏、脑、头肾中表达量极低(几乎检测不到)。在低磷饲料中添加维生素D3显著诱导了该基因的上调表达。研究结果展示了黄颡鱼MHC class Ⅱ抗原基因的分子结构、组织表达以及维生素D3的作用,在降低磷排放的同时,为今后黄颡鱼免疫抗病及分子选育等方向的深入研究及免疫型饲料的使用奠定了基础。展开更多
目的:构建携带小鼠MHCII类分子反式激活因子(MHC class II molecule transactivator,CIITA)突变体基因的腺病毒,并观察腺病毒介导该基因的表达情况以及表达产物的体外功能。方法:采用常规分子生物学方法从IFN-γ诱导后的BALB/c小鼠腹腔...目的:构建携带小鼠MHCII类分子反式激活因子(MHC class II molecule transactivator,CIITA)突变体基因的腺病毒,并观察腺病毒介导该基因的表达情况以及表达产物的体外功能。方法:采用常规分子生物学方法从IFN-γ诱导后的BALB/c小鼠腹腔巨噬细胞中获得IV型CIITA cDNA;利用重叠延伸PCR法构建CIITA突变体基因,并克隆入表达载体pIRES;采用pAdEasy-1系统获得具有感染能力的、携带CIITA突变体基因的缺陷型重组腺病毒(Ad-CIITAm)和空载对照病毒(Ad-GFP),并经大量扩增、纯化及滴度测定;将Ad-CIITAm和Ad-GFP分别感染HeLa细胞和Raji细胞,流式细胞术观察对诱导型和组成型HLA-DR分子表达的影响。结果:成功克隆了小鼠CIITA突变体基因,并构建了携带小鼠CIITA突变体基因的重组腺病毒Ad-CIITAm;经流式细胞术证实感染Ad-CIITAm的Hela和Raji细胞较感染Ad-GFP的细胞,其表面HLA-DR分子的表达均受到明显的抑制。结论:本实验证实了重组腺病毒介导表达的小鼠CIITA突变体在体外能够有效地抑制MHCII类分子的表达。展开更多
文摘Membrane-associated RING-CH-1 (MARCH1) is an E3 ubiquitin ligase which targets MHC-II, CD86 and various other molecules for degradation. It is one of the most efficient post-translational regulators of antigen presentation. MARCH1 is expressed in resting immature dendritic cells and B cells but can be induced in other cell types. While activation of most immune cells results in a reduction in MARCH1 gene expression, its anti-inflammatory properties are highlighted by its induction in response to IL-10. Here, we review what is known about the regulation of MARCH1 gene expression in response to IL-10 by various immune cells. We speculate on the role of MARCH1 ininfection, its differential expression pattern and the promise that this E3 ubiquitin ligase holds to influence immune responses and mitigate immune pathologies.
文摘为进一步丰富鱼类MHC class Ⅱ基因的研究,同时也为进一步探讨低磷饲料中添加维生素D3对鱼类免疫功能可能的影响,实验利用RACE(Rapid-amplification of c DNA ends)即c DNA末端快速扩增技术,成功克隆出黄颡鱼(Pelteobagrus fulvidraco)主要组织相容性复合体(Major histocompatibility complex,MHC)class Ⅱ抗原基因,全长1074 bp,其中ORF(Open reading frame)708 bp,编码236个氨基酸,5′UTR(5′端非翻译区)78 bp,3′UTR(3′端非翻译区)259 bp。进行氨基酸序列比对分析得到:黄颡鱼MHC class Ⅱ基因ORF氨基酸序列与长吻逘(Leiocassis longirostris)的氨基酸序列相似度最高为69.5%,与锦鲤(Cyprinus carpio)的氨基酸序列相似度最低为50.4%。利用q PCR对黄颡鱼MHC class Ⅱ基因进行组织表达分析,结果表明MHC class Ⅱ在小肠、肝脏、鳃中表达较高;在肌肉、鳍条中表达较低;而在肾、脾脏、脑、头肾中表达量极低(几乎检测不到)。在低磷饲料中添加维生素D3显著诱导了该基因的上调表达。研究结果展示了黄颡鱼MHC class Ⅱ抗原基因的分子结构、组织表达以及维生素D3的作用,在降低磷排放的同时,为今后黄颡鱼免疫抗病及分子选育等方向的深入研究及免疫型饲料的使用奠定了基础。
文摘目的:构建携带小鼠MHCII类分子反式激活因子(MHC class II molecule transactivator,CIITA)突变体基因的腺病毒,并观察腺病毒介导该基因的表达情况以及表达产物的体外功能。方法:采用常规分子生物学方法从IFN-γ诱导后的BALB/c小鼠腹腔巨噬细胞中获得IV型CIITA cDNA;利用重叠延伸PCR法构建CIITA突变体基因,并克隆入表达载体pIRES;采用pAdEasy-1系统获得具有感染能力的、携带CIITA突变体基因的缺陷型重组腺病毒(Ad-CIITAm)和空载对照病毒(Ad-GFP),并经大量扩增、纯化及滴度测定;将Ad-CIITAm和Ad-GFP分别感染HeLa细胞和Raji细胞,流式细胞术观察对诱导型和组成型HLA-DR分子表达的影响。结果:成功克隆了小鼠CIITA突变体基因,并构建了携带小鼠CIITA突变体基因的重组腺病毒Ad-CIITAm;经流式细胞术证实感染Ad-CIITAm的Hela和Raji细胞较感染Ad-GFP的细胞,其表面HLA-DR分子的表达均受到明显的抑制。结论:本实验证实了重组腺病毒介导表达的小鼠CIITA突变体在体外能够有效地抑制MHCII类分子的表达。
