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
目的探讨循环可溶性MHC-I类链相关蛋白A[soluble major histocompatibility complex class I-related chain A,sMICA)、可溶性MHC-I类链相关蛋白B(soluble major histocompatibility complex class I-related chain B,sMICB]与系统性红...目的探讨循环可溶性MHC-I类链相关蛋白A[soluble major histocompatibility complex class I-related chain A,sMICA)、可溶性MHC-I类链相关蛋白B(soluble major histocompatibility complex class I-related chain B,sMICB]与系统性红斑狼疮(systemic lupus erythematosus,SLE)疾病活动性、自身抗体的关系。方法选择2020年1月~2023年1月重庆大学附属黔江医院收治的156例SLE患者(SLE组)和门诊体检中心体检的103例健康志愿者(对照组)。根据SLE疾病活动度评分(SLE disease activity index,SLEDAI)将SLE患者分为轻度活动组(n=43)、中度活动组(n=69)和重度活动组(n=44)。检测血清sMICA,sMICB水平以及自身抗体、外周血NK细胞占比,Spearman或Pearson分析sMICA,sMICB与评分、自身抗体、外周血NK细胞占比的相关性,受试者工作特征(ROC)曲线用来分析sMICA和sMICB诊断SLE活动度的价值。结果SLE组血清sMICA(173.65±23.92 pg/ml),sMICB(96.35±15.74 pg/ml)水平高于对照组(32.51±6.27 pg/ml,12.03±2.47 pg/ml),外周血CD3^(-)CD56^(+)NK细胞(12.02%±2.65%)占比低于对照组(18.35%±3.71%),差异具有统计学意义(t=58.498,53.897,-16.010,均P<0.05)。重度活动组血清sMICA,sMICB水平高于中度活动组和轻度活动组(t=8.192,12.352;19.652,23.742,均P<0.05),外周血CD3^(-)CD56^(+)NK细胞占比低于中度活动组和轻度活动组(t=8.154,10.658,均P<0.05),差异具有统计学意义。不同疾病活动SLE患者抗‐dsDNA抗体、抗核抗体、抗核小体抗体和抗组蛋白抗体阳性率比较,差异具有统计学意义(χ^(2)=8.795,7.216,7.539,8.946,均P<0.05)。SLE患者血清sMICA,sMICB水平与SLEDAI评分、抗‐dsDNA抗体、抗核抗体、抗核小体抗体、抗组蛋白抗体呈正相关(r=0.206~0.402,均P<0.05),与外周血CD3^(-)CD56^(+)NK细胞占比呈负相关(r=-0.563,-0.427,均P<0.05)。sMICA和sMICB诊断SLE重度活动的曲线下面积为0.652,0.704,联合sMICA,sMICB诊断SLE重度活动的曲线下面积为0.812,高于单独诊断(Z=3.050,2.346,均P<0.05)。结论SLE患者血清sMICA和sMICB水平增高,且与SLE自身抗体阳性率增加、外周血NK细胞占比降低、疾病活动性增强有关,可作为SLE的潜在标志物。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。
