Background In addition to the well-known antibodies against human leukocyte antigens （HLA）-induced kidney-graft rejection, polymorphic major-histocompatibility-complex （MHC） class Ⅰ-related chain A （MICA） antig...Background In addition to the well-known antibodies against human leukocyte antigens （HLA）-induced kidney-graft rejection, polymorphic major-histocompatibility-complex （MHC） class Ⅰ-related chain A （MICA） antigens can elicit antibodies and have been suggested to play a role in the antibody-mediated allograft rejection （AMR）. We carded out a prospective study of MICA antibodies in post-renal transplant patients to determine the association between MICA antibodies, C4d staining, histological features, and graft outcome.Methods We tested 52 patients who had biopsy results due to graft dysfunction. The MICA antibodies in concurrent sera were determined by Luminex. All patients were followed up for one year after renal biopsy. The influence of antibody production on the function of graft was analyzed.Results Antibodies against MICA were positive in 15 out of the 52 patients （28.9%）. The presence of MICA antibodies was associated with renal-allograft deterioration. During one-year follow-up, the estimated glomerular filtration rate （eGFR） decreased （24.0±3.4）% among recipients with anti-MICA antibodies. However, among recipients without anti-MICA antibodies, the eGFR has declined only （8.4＋3.0）% （P=0.017）. The association between C4d staining,histological features and MICA antibody production was found no significant difference.Conclusion Besides anti-HLA antibodies, the presence of post-transplant MICA antibody is associated with poor graft outcome and increases the risk of graft failure.展开更多
目的构建人类MHC-Ⅰ类链相关基因A(MICA)的真核表达载体,转染人舌鳞癌脑高转移Tca8113-Tb细胞,建立稳定过表达MICA基因的口腔鳞癌细胞系。方法采用PCR技术扩增pCMV-SPORT6-MICA中编码MICA基因的cDNA序列,重组至有绿色荧光蛋白标记的真...目的构建人类MHC-Ⅰ类链相关基因A(MICA)的真核表达载体,转染人舌鳞癌脑高转移Tca8113-Tb细胞,建立稳定过表达MICA基因的口腔鳞癌细胞系。方法采用PCR技术扩增pCMV-SPORT6-MICA中编码MICA基因的cDNA序列,重组至有绿色荧光蛋白标记的真核表达载体pEGFP-N1,构建最终的表达载体pEGFP-N1-MICA,脂质体法转染Tca8113-Tb细胞,G418筛选,荧光显微镜下观察绿色荧光蛋白的表达,有限稀释法建立稳定过表达MICA基因的Tca8113-Tb细胞系,RT-PCR、real time PCR和免疫细胞化学检测MICA在该细胞中的表达。结果通过PCR技术获取了MICA基因并成功克隆入载体,测序鉴定该序列与GenBank中的序列相同。转染的细胞可见绿色荧光蛋白表达,RT-PCR、real time PCR及免疫细胞化学检测到目的基因MICA在转染细胞中为过表达。结论 pEGFP-N1-MICA真核表达载体的成功构建与稳定转染Tca8113-Tb细胞系的建立,为进一步研究该基因的功能奠定了良好的实验基础。展开更多
目的:探讨胃癌细胞系MKN中HLA-Ⅰ类抗原表达水平异常的分子机制。方法:以胃癌细胞系BGC823、MGC803和SGC7901为对照,利用流式细胞仪检测细胞表面HLA-Ⅰ类复合物的表达情况,W estern b lot检测HLA-Ⅰ类分子重链及轻链表达情况,半定量RT-...目的:探讨胃癌细胞系MKN中HLA-Ⅰ类抗原表达水平异常的分子机制。方法:以胃癌细胞系BGC823、MGC803和SGC7901为对照,利用流式细胞仪检测细胞表面HLA-Ⅰ类复合物的表达情况,W estern b lot检测HLA-Ⅰ类分子重链及轻链表达情况,半定量RT-PCR检测HLA-Ⅰ类分子重链、轻链和抗原加工分子mRNA水平的表达情况,并利用HLA基因区域的微卫星序列检测MKN细胞HLA基因在DNA水平的完整性。结果:胃癌细胞系MKN表面HLA-Ⅰ类复合物表达降低,重链A及B/C位点蛋白无表达而轻链表达无异常。在mRNA水平,胃癌细胞系MKN存在重链A、B、C各位点和抗原加工分子TAP1、LMP2、Tapasin、PA28β的表达缺失。微卫星DNA扩增结果初步显示,MKN细胞无HLA基因区域DNA的大片段丢失。结论:胃癌细胞系MKN HLA-Ⅰ类分子复合物表达降低可能是由HLA-Ⅰ类分子重链及其加工分子在转录水平改变而引起的。展开更多
Olfactory ensheathing glia promote axonal regeneration in the mammalian central nervous system,including retinal ganglion cell axonal growth through the injured optic nerve.Still,it is unknown whether olfactory enshea...Olfactory ensheathing glia promote axonal regeneration in the mammalian central nervous system,including retinal ganglion cell axonal growth through the injured optic nerve.Still,it is unknown whether olfactory ensheathing glia also have neuroprotective properties.Olfactory ensheathing glia express brain-derived neurotrophic factor,one of the best neuroprotectants for axotomized retinal ganglion cells.Therefore,we aimed to investigate the neuroprotective capacity of olfactory ensheating glia after optic nerve crush.Olfactory ensheathing glia cells from an established rat immortalized clonal cell line,TEG3,were intravitreally injected in intact and axotomized retinas in syngeneic and allogeneic mode with or without microglial inhibition or immunosuppressive treatments.Anatomical and gene expression analyses were performed.Olfactory bulb-derived primary olfactory ensheathing glia and TEG3 express major histocompatibility complex classⅡmolecules.Allogeneically and syngenically transplanted TEG3 cells survived in the vitreous for up to 21 days,forming an epimembrane.In axotomized retinas,only the allogeneic TEG3 transplant rescued retinal ganglion cells at 7 days but not at 21 days.In these retinas,microglial anatomical activation was higher than after optic nerve crush alone.In intact retinas,both transplants activated microglial cells and caused retinal ganglion cell death at 21 days,a loss that was higher after allotransplantation,triggered by pyroptosis and partially rescued by microglial inhibition or immunosuppression.However,neuroprotection of axotomized retinal ganglion cells did not improve with these treatments.The different neuroprotective properties,different toxic effects,and different responses to microglial inhibitory treatments of olfactory ensheathing glia in the retina depending on the type of transplant highlight the importance of thorough preclinical studies to explore these variables.展开更多
文摘Background In addition to the well-known antibodies against human leukocyte antigens （HLA）-induced kidney-graft rejection, polymorphic major-histocompatibility-complex （MHC） class Ⅰ-related chain A （MICA） antigens can elicit antibodies and have been suggested to play a role in the antibody-mediated allograft rejection （AMR）. We carded out a prospective study of MICA antibodies in post-renal transplant patients to determine the association between MICA antibodies, C4d staining, histological features, and graft outcome.Methods We tested 52 patients who had biopsy results due to graft dysfunction. The MICA antibodies in concurrent sera were determined by Luminex. All patients were followed up for one year after renal biopsy. The influence of antibody production on the function of graft was analyzed.Results Antibodies against MICA were positive in 15 out of the 52 patients （28.9%）. The presence of MICA antibodies was associated with renal-allograft deterioration. During one-year follow-up, the estimated glomerular filtration rate （eGFR） decreased （24.0±3.4）% among recipients with anti-MICA antibodies. However, among recipients without anti-MICA antibodies, the eGFR has declined only （8.4＋3.0）% （P=0.017）. The association between C4d staining,histological features and MICA antibody production was found no significant difference.Conclusion Besides anti-HLA antibodies, the presence of post-transplant MICA antibody is associated with poor graft outcome and increases the risk of graft failure.
文摘目的构建人类MHC-Ⅰ类链相关基因A(MICA)的真核表达载体,转染人舌鳞癌脑高转移Tca8113-Tb细胞,建立稳定过表达MICA基因的口腔鳞癌细胞系。方法采用PCR技术扩增pCMV-SPORT6-MICA中编码MICA基因的cDNA序列,重组至有绿色荧光蛋白标记的真核表达载体pEGFP-N1,构建最终的表达载体pEGFP-N1-MICA,脂质体法转染Tca8113-Tb细胞,G418筛选,荧光显微镜下观察绿色荧光蛋白的表达,有限稀释法建立稳定过表达MICA基因的Tca8113-Tb细胞系,RT-PCR、real time PCR和免疫细胞化学检测MICA在该细胞中的表达。结果通过PCR技术获取了MICA基因并成功克隆入载体,测序鉴定该序列与GenBank中的序列相同。转染的细胞可见绿色荧光蛋白表达,RT-PCR、real time PCR及免疫细胞化学检测到目的基因MICA在转染细胞中为过表达。结论 pEGFP-N1-MICA真核表达载体的成功构建与稳定转染Tca8113-Tb细胞系的建立,为进一步研究该基因的功能奠定了良好的实验基础。
文摘目的:探讨胃癌细胞系MKN中HLA-Ⅰ类抗原表达水平异常的分子机制。方法:以胃癌细胞系BGC823、MGC803和SGC7901为对照,利用流式细胞仪检测细胞表面HLA-Ⅰ类复合物的表达情况,W estern b lot检测HLA-Ⅰ类分子重链及轻链表达情况,半定量RT-PCR检测HLA-Ⅰ类分子重链、轻链和抗原加工分子mRNA水平的表达情况,并利用HLA基因区域的微卫星序列检测MKN细胞HLA基因在DNA水平的完整性。结果:胃癌细胞系MKN表面HLA-Ⅰ类复合物表达降低,重链A及B/C位点蛋白无表达而轻链表达无异常。在mRNA水平,胃癌细胞系MKN存在重链A、B、C各位点和抗原加工分子TAP1、LMP2、Tapasin、PA28β的表达缺失。微卫星DNA扩增结果初步显示,MKN细胞无HLA基因区域DNA的大片段丢失。结论:胃癌细胞系MKN HLA-Ⅰ类分子复合物表达降低可能是由HLA-Ⅰ类分子重链及其加工分子在转录水平改变而引起的。
基金supported by the Spanish Ministry of Economy and Competitiveness,No.PID2019-106498GB-I00(to MVS)the Instituto de Salud CarlosⅢ,Fondo Europeo de Desarrollo Regional“Una manera de hacer Europa”,No.PI19/00071(to MAB)+1 种基金Ministerio de Ciencia e Innovación Project,No.SAF2017-82736-C2-1-R(to MTMF)in Universidad Autónoma de MadridFundación Universidad Francisco de Vitoria(to JS)。
文摘Olfactory ensheathing glia promote axonal regeneration in the mammalian central nervous system,including retinal ganglion cell axonal growth through the injured optic nerve.Still,it is unknown whether olfactory ensheathing glia also have neuroprotective properties.Olfactory ensheathing glia express brain-derived neurotrophic factor,one of the best neuroprotectants for axotomized retinal ganglion cells.Therefore,we aimed to investigate the neuroprotective capacity of olfactory ensheating glia after optic nerve crush.Olfactory ensheathing glia cells from an established rat immortalized clonal cell line,TEG3,were intravitreally injected in intact and axotomized retinas in syngeneic and allogeneic mode with or without microglial inhibition or immunosuppressive treatments.Anatomical and gene expression analyses were performed.Olfactory bulb-derived primary olfactory ensheathing glia and TEG3 express major histocompatibility complex classⅡmolecules.Allogeneically and syngenically transplanted TEG3 cells survived in the vitreous for up to 21 days,forming an epimembrane.In axotomized retinas,only the allogeneic TEG3 transplant rescued retinal ganglion cells at 7 days but not at 21 days.In these retinas,microglial anatomical activation was higher than after optic nerve crush alone.In intact retinas,both transplants activated microglial cells and caused retinal ganglion cell death at 21 days,a loss that was higher after allotransplantation,triggered by pyroptosis and partially rescued by microglial inhibition or immunosuppression.However,neuroprotection of axotomized retinal ganglion cells did not improve with these treatments.The different neuroprotective properties,different toxic effects,and different responses to microglial inhibitory treatments of olfactory ensheathing glia in the retina depending on the type of transplant highlight the importance of thorough preclinical studies to explore these variables.
