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
Background Humoral immunity is an important factor for long-term survival of renal allograft. Here we performed a four-year follow-up to explore the clinical significance of monitoring anti-human leukocyte antigens （...Background Humoral immunity is an important factor for long-term survival of renal allograft. Here we performed a four-year follow-up to explore the clinical significance of monitoring anti-human leukocyte antigens （HLA） and anti-major histocompatibility complex class I-related chain A （MICA） antibody expression after kidney transplantation. Methods We obtained serial serum samples from 84 kidney transplant patients over a four-year period. All patients were followed up at least 6 months after transplantation and had at least two follow-up points. Anti-HLA and anti-MICA antibody titres and serum creatinine （SCr） levels were evaluated at each follow-up. Patients were divided into 4 groups： HLA（＋） MICA（-）, HLA（-）MICA（＋）, HLA（＋）MICA（＋） and HLA（-）MICA（-）. The impact of post-transplant antibody level on kidney allograft function was evaluated. Results Antibodies were detected in 38.1% （32/84） of the renal allograft recipients. HLA, MICA and HLA＋MICA expression was observed in 18.89%, 14.44% and 5.93% of the recipients respectively. The most frequent anti-HLA and anti-MICA specific antibodies identified were All, A24, A29, A32, A33, A80; B7, B13, B37; DR17, DR12, DR18, DR52, DR53, DR1, DR4, DR9, DR51; DQ7, DQ4, DQ8, DQ2, DQ9, DQ5, DQ6 and MICA02, MICA18, MICA19, MICA07, MICA27. As the time after transplantation elapsed, more recipients developed de novo antibody expression. Total 11.91% （10/84） of the recipients had de novo antibody expression during the follow up. The average level of SCr and the percentage of recipients with abnormal allograft function were significantly higher in recipients with anti-HLA and/or anti- MICA antibody expression than those without. The appearance of anti-HLA and anti-MICA antibody expression always preceded the increase in SCr value. Conclusions Anti-HLA and anti-MICA antibody expression has predictive value for early and late allograft dysfunction. The presence of donor specific antibody is detrimental to graft function and graft survival.展开更多
Objective To review the role of polymorphism of major histocompatibility complex class I-related chain A （MICA） gene and antibodies against MICA antigens in transplant immunology. Data sources The data used in this ...Objective To review the role of polymorphism of major histocompatibility complex class I-related chain A （MICA） gene and antibodies against MICA antigens in transplant immunology. Data sources The data used in this review were mainly from our own results and from the relevant English language literatures published from 1999 to 2010. Some data presented in this review are in press.Study selection Articles regarding MICA gene discovery and pioneering finding of antibodies against MICA antigen and allograft rejection were selected. This review chronicles the development of our understanding of the role that MICA antigens and antibodies may play in organ transplantation.Results Polymorphic glycoprotein MICA antigens were detected on freshly isolated human umbilical cord endothelial cells, but not on peripheral lymphocytes. Antibodies were found and typing of recipients and donors by sequencing the MICA alleles has established that de novo antibodies produced in kidney transplant recipients are directed at mismatched MICA epitopes and are associated with acute rejection and chronic transplant failure. The specificity of antibodies against the epitopes of MICA antigens were well characterized by donor MICA typing, single antigen array testing with antibody absorption and elution. Acute graft-versus-host disease was observed in stem-cell recipients who were mismatched for MICA.Conclusions Immunization against mismatched MICA epitopes encountered in donor organs after transplantation may result in antibodies against MICA alleles. Testing for MICA donor-specific antibodies （DSA） which are associated with early failure of kidney transplants may be helpful for identifying some of the targets of antibodies against antigens other than the human leukocyte antigen （HLA） and for improving transplantation outcome.展开更多
Background Sensitized recipients have a high risk of immunological graft loss due to hyperacute rejection and/or accelerated acute rejection. The presence of major histocompatibility complex class I-related chain A （...Background Sensitized recipients have a high risk of immunological graft loss due to hyperacute rejection and/or accelerated acute rejection. The presence of major histocompatibility complex class I-related chain A （MICA） antibodies has also been described associated with an increased rate of kidney-allograft rejection. The aim of this study was to describe the expression of MICA antibodies in sensitized recipients of renal transplantation and evaluate its influence on the kidney transplantation recipients.Methods A total of 29 sensitized recipients were included in this study. All patients received the MICA antibodies detection before and after protein A immunoadsorption. Panel reactive antibody （PRA）, HLA-matches, acute rejection and postoperative one to four-week serum creatinine level were also collected and analyzed, respectively. No prisoners were used in this study.Results Eight patients （27.6%） in all 29 sensitized recipients expressed the MICA antibodies but did not show higher acute rejection rate than the non-expressed patients （3/8, 37.5% vs. 8/21, 38.1%; P=1.000）. Recipients with PRA 〉40% showed higher expression levels of MICA antibodies than the recipients with PRA 〈40% （7/16, 43.8% vs. 1/13, 8.3%; P=0.044）. HLA mismatch did not have any effect on the expression of MICA antibodies （P=1.000）. MICA antibodies positive group had higher serum creatinine level than the control in postoperative one week （（135.4±21.4） μmol/L vs. （108.6±31.6） μmol/L, P=0.036）, but no significant difference in postoperative four weeks （（89.0±17.1） μmol/L vs. （77.1±15.9） μmol/L, P=0.089）. MICA antibodies decreased significantly after protein A immunoadsorption.Conclusions MICA antibodies increase in the sensitized recipients, which have significant effects on the function of aliograft in early postoperative period. Protein A immunoadsorption can decrease MICA antibodies effectively in sensitized recipients.展开更多
Proper understanding of global distribution of infectious diseases is an important part of disease management and policy making. However, data are subject to complexities caused by heterogeneities across host classes ...Proper understanding of global distribution of infectious diseases is an important part of disease management and policy making. However, data are subject to complexities caused by heterogeneities across host classes and space-time epidemic processes. This paper seeks to suggest or propose Bayesian spatio-temporal model for modeling and mapping tuberculosis relative risks in space and time as well identify risks factors associated with the tuberculosis and counties in Kenya with high tuberculosis relative risks. In this paper, we used spatio-temporal Bayesian hierarchical models to study the pattern of tuberculosis relative risks in Kenya. The Markov Chain Monte Carlo method via WinBUGS and R packages were used for simulations and estimation of the parameter estimates. The best fitting model is selected using the Deviance Information Criterion proposed by Spiegelhalter and colleagues. Among the spatio-temporal models used, the Knorr-Held model with space-time interaction type III and IV fit the data well but type IV appears better than type III. Variation in tuberculosis risk is observed among Kenya counties and clustering among counties with high tuberculosis relative risks. The prevalence of HIV is identified as the determinant of TB. We found clustering and heterogeneity of TB risk among high rate counties and the overall tuberculosis risk is slightly decreasing from 2002-2009. We proposed that the Knorr-Held model with interaction type IV should be used to model and map Kenyan tuberculosis relative risks. Interaction of TB relative risk in space and time increases among rural counties that share boundaries with urban counties with high tuberculosis risk. This is due to the ability of models to borrow strength from neighboring counties, such that nearby counties have similar risk. Although the approaches are less than ideal, we hope that our study provide a useful stepping stone in the development of spatial and spatio-temporal methodology for the statistical analysis of risk from tuberculosis in Kenya.展开更多
目的探讨骨肉瘤组织中MHCⅠ类链相关蛋白A(MHC class Ⅰ chain-related A,MICA)、MMP-9和NF-κB的表达及相互间的关系,为研究骨肉瘤组织中MICA蛋白表达和脱落机制提供组织学依据。方法应用免疫组织化学方法检测66例骨肉瘤、11例骨母细胞...目的探讨骨肉瘤组织中MHCⅠ类链相关蛋白A(MHC class Ⅰ chain-related A,MICA)、MMP-9和NF-κB的表达及相互间的关系,为研究骨肉瘤组织中MICA蛋白表达和脱落机制提供组织学依据。方法应用免疫组织化学方法检测66例骨肉瘤、11例骨母细胞瘤,8例骨化性纤维瘤和6例正常骨组织中MICA蛋白的表达情况,并检测66例骨肉瘤组织中MMP-9和NF-κB蛋白的表达情况。结果(1)MICA蛋白在骨肉瘤组织、骨母细胞瘤、骨化性纤维瘤和正常骨组织的表达率分别为51.6%(34/66)、9%(1/11)、0(0/8)、0(0/6),骨肉瘤组织中MICA表达与骨母细胞瘤(P=0.01)、骨化性纤维瘤(P<0.01)和正常骨组织(P<0.05的差异有显著性。(2)骨肉瘤组织中MMP-9和NF-κB表达率分别为55%(36/66)和73%(48/66);NF-κB与MICA(r=0.373,P<0.01)和MMP-9(r=0.536,P<0.01)的表达呈正相关关系。结论MICA蛋白可作为骨肉瘤诊断的分子标志物;NF-κB可能参与MICA蛋白表达和脱落的分子机制,NF-κB可作为骨肉瘤免疫治疗的候选靶点。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】检测骨肉瘤组织中MHCI类链相关蛋白A(MHC class I chain-related A,MICA)基因mRNA和蛋白的表达情况,为探讨骨肉瘤的肿瘤免疫机制提供信息。【方法】应用RT-PCR方法检测MICA mRNA在11例新鲜骨肉瘤组织和5个骨肉瘤细胞株中的表达...【目的】检测骨肉瘤组织中MHCI类链相关蛋白A(MHC class I chain-related A,MICA)基因mRNA和蛋白的表达情况,为探讨骨肉瘤的肿瘤免疫机制提供信息。【方法】应用RT-PCR方法检测MICA mRNA在11例新鲜骨肉瘤组织和5个骨肉瘤细胞株中的表达情况;分别应用免疫组织化学方法、Western blot和流式细胞术检测66例石蜡包埋骨肉瘤组织、6例石蜡包埋正常人骨组织、9例新鲜骨肉瘤组织和5个骨肉瘤细胞株中MICA蛋白的表达情况。【结果】骨肉瘤组织中MICA mRNA表达率为81.8%(9/11),5个骨肉瘤细胞株均表达MICA mRNA;MICA蛋白在骨肉瘤组织和正常骨组织的表达率分别为51.6%(34/66)和0%(0/6);骨肉瘤细胞株Saos-2、MG63和HOS阳性表达膜表面MICA蛋白,而细胞株U2OS和OS732只有少量细胞呈阳性表达。【结论】MICA mRNA和蛋白广泛表达于骨肉瘤组织和细胞株。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.
基金the grants from the the National Science Foundation of China，the Key Discipline of Medicine of Jiangsu Province，the Outstanding Medical Academic Leader Program of Jiangsu Province，the Science Foundation of Jiangsu Province，the Key Laboratory Foundation of Suzhou
文摘Background Humoral immunity is an important factor for long-term survival of renal allograft. Here we performed a four-year follow-up to explore the clinical significance of monitoring anti-human leukocyte antigens （HLA） and anti-major histocompatibility complex class I-related chain A （MICA） antibody expression after kidney transplantation. Methods We obtained serial serum samples from 84 kidney transplant patients over a four-year period. All patients were followed up at least 6 months after transplantation and had at least two follow-up points. Anti-HLA and anti-MICA antibody titres and serum creatinine （SCr） levels were evaluated at each follow-up. Patients were divided into 4 groups： HLA（＋） MICA（-）, HLA（-）MICA（＋）, HLA（＋）MICA（＋） and HLA（-）MICA（-）. The impact of post-transplant antibody level on kidney allograft function was evaluated. Results Antibodies were detected in 38.1% （32/84） of the renal allograft recipients. HLA, MICA and HLA＋MICA expression was observed in 18.89%, 14.44% and 5.93% of the recipients respectively. The most frequent anti-HLA and anti-MICA specific antibodies identified were All, A24, A29, A32, A33, A80; B7, B13, B37; DR17, DR12, DR18, DR52, DR53, DR1, DR4, DR9, DR51; DQ7, DQ4, DQ8, DQ2, DQ9, DQ5, DQ6 and MICA02, MICA18, MICA19, MICA07, MICA27. As the time after transplantation elapsed, more recipients developed de novo antibody expression. Total 11.91% （10/84） of the recipients had de novo antibody expression during the follow up. The average level of SCr and the percentage of recipients with abnormal allograft function were significantly higher in recipients with anti-HLA and/or anti- MICA antibody expression than those without. The appearance of anti-HLA and anti-MICA antibody expression always preceded the increase in SCr value. Conclusions Anti-HLA and anti-MICA antibody expression has predictive value for early and late allograft dysfunction. The presence of donor specific antibody is detrimental to graft function and graft survival.
文摘Objective To review the role of polymorphism of major histocompatibility complex class I-related chain A （MICA） gene and antibodies against MICA antigens in transplant immunology. Data sources The data used in this review were mainly from our own results and from the relevant English language literatures published from 1999 to 2010. Some data presented in this review are in press.Study selection Articles regarding MICA gene discovery and pioneering finding of antibodies against MICA antigen and allograft rejection were selected. This review chronicles the development of our understanding of the role that MICA antigens and antibodies may play in organ transplantation.Results Polymorphic glycoprotein MICA antigens were detected on freshly isolated human umbilical cord endothelial cells, but not on peripheral lymphocytes. Antibodies were found and typing of recipients and donors by sequencing the MICA alleles has established that de novo antibodies produced in kidney transplant recipients are directed at mismatched MICA epitopes and are associated with acute rejection and chronic transplant failure. The specificity of antibodies against the epitopes of MICA antigens were well characterized by donor MICA typing, single antigen array testing with antibody absorption and elution. Acute graft-versus-host disease was observed in stem-cell recipients who were mismatched for MICA.Conclusions Immunization against mismatched MICA epitopes encountered in donor organs after transplantation may result in antibodies against MICA alleles. Testing for MICA donor-specific antibodies （DSA） which are associated with early failure of kidney transplants may be helpful for identifying some of the targets of antibodies against antigens other than the human leukocyte antigen （HLA） and for improving transplantation outcome.
文摘Background Sensitized recipients have a high risk of immunological graft loss due to hyperacute rejection and/or accelerated acute rejection. The presence of major histocompatibility complex class I-related chain A （MICA） antibodies has also been described associated with an increased rate of kidney-allograft rejection. The aim of this study was to describe the expression of MICA antibodies in sensitized recipients of renal transplantation and evaluate its influence on the kidney transplantation recipients.Methods A total of 29 sensitized recipients were included in this study. All patients received the MICA antibodies detection before and after protein A immunoadsorption. Panel reactive antibody （PRA）, HLA-matches, acute rejection and postoperative one to four-week serum creatinine level were also collected and analyzed, respectively. No prisoners were used in this study.Results Eight patients （27.6%） in all 29 sensitized recipients expressed the MICA antibodies but did not show higher acute rejection rate than the non-expressed patients （3/8, 37.5% vs. 8/21, 38.1%; P=1.000）. Recipients with PRA 〉40% showed higher expression levels of MICA antibodies than the recipients with PRA 〈40% （7/16, 43.8% vs. 1/13, 8.3%; P=0.044）. HLA mismatch did not have any effect on the expression of MICA antibodies （P=1.000）. MICA antibodies positive group had higher serum creatinine level than the control in postoperative one week （（135.4±21.4） μmol/L vs. （108.6±31.6） μmol/L, P=0.036）, but no significant difference in postoperative four weeks （（89.0±17.1） μmol/L vs. （77.1±15.9） μmol/L, P=0.089）. MICA antibodies decreased significantly after protein A immunoadsorption.Conclusions MICA antibodies increase in the sensitized recipients, which have significant effects on the function of aliograft in early postoperative period. Protein A immunoadsorption can decrease MICA antibodies effectively in sensitized recipients.
文摘Proper understanding of global distribution of infectious diseases is an important part of disease management and policy making. However, data are subject to complexities caused by heterogeneities across host classes and space-time epidemic processes. This paper seeks to suggest or propose Bayesian spatio-temporal model for modeling and mapping tuberculosis relative risks in space and time as well identify risks factors associated with the tuberculosis and counties in Kenya with high tuberculosis relative risks. In this paper, we used spatio-temporal Bayesian hierarchical models to study the pattern of tuberculosis relative risks in Kenya. The Markov Chain Monte Carlo method via WinBUGS and R packages were used for simulations and estimation of the parameter estimates. The best fitting model is selected using the Deviance Information Criterion proposed by Spiegelhalter and colleagues. Among the spatio-temporal models used, the Knorr-Held model with space-time interaction type III and IV fit the data well but type IV appears better than type III. Variation in tuberculosis risk is observed among Kenya counties and clustering among counties with high tuberculosis relative risks. The prevalence of HIV is identified as the determinant of TB. We found clustering and heterogeneity of TB risk among high rate counties and the overall tuberculosis risk is slightly decreasing from 2002-2009. We proposed that the Knorr-Held model with interaction type IV should be used to model and map Kenyan tuberculosis relative risks. Interaction of TB relative risk in space and time increases among rural counties that share boundaries with urban counties with high tuberculosis risk. This is due to the ability of models to borrow strength from neighboring counties, such that nearby counties have similar risk. Although the approaches are less than ideal, we hope that our study provide a useful stepping stone in the development of spatial and spatio-temporal methodology for the statistical analysis of risk from tuberculosis in Kenya.
文摘目的探讨骨肉瘤组织中MHCⅠ类链相关蛋白A(MHC class Ⅰ chain-related A,MICA)、MMP-9和NF-κB的表达及相互间的关系,为研究骨肉瘤组织中MICA蛋白表达和脱落机制提供组织学依据。方法应用免疫组织化学方法检测66例骨肉瘤、11例骨母细胞瘤,8例骨化性纤维瘤和6例正常骨组织中MICA蛋白的表达情况,并检测66例骨肉瘤组织中MMP-9和NF-κB蛋白的表达情况。结果(1)MICA蛋白在骨肉瘤组织、骨母细胞瘤、骨化性纤维瘤和正常骨组织的表达率分别为51.6%(34/66)、9%(1/11)、0(0/8)、0(0/6),骨肉瘤组织中MICA表达与骨母细胞瘤(P=0.01)、骨化性纤维瘤(P<0.01)和正常骨组织(P<0.05的差异有显著性。(2)骨肉瘤组织中MMP-9和NF-κB表达率分别为55%(36/66)和73%(48/66);NF-κB与MICA(r=0.373,P<0.01)和MMP-9(r=0.536,P<0.01)的表达呈正相关关系。结论MICA蛋白可作为骨肉瘤诊断的分子标志物;NF-κB可能参与MICA蛋白表达和脱落的分子机制,NF-κB可作为骨肉瘤免疫治疗的候选靶点。
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】检测骨肉瘤组织中MHCI类链相关蛋白A(MHC class I chain-related A,MICA)基因mRNA和蛋白的表达情况,为探讨骨肉瘤的肿瘤免疫机制提供信息。【方法】应用RT-PCR方法检测MICA mRNA在11例新鲜骨肉瘤组织和5个骨肉瘤细胞株中的表达情况;分别应用免疫组织化学方法、Western blot和流式细胞术检测66例石蜡包埋骨肉瘤组织、6例石蜡包埋正常人骨组织、9例新鲜骨肉瘤组织和5个骨肉瘤细胞株中MICA蛋白的表达情况。【结果】骨肉瘤组织中MICA mRNA表达率为81.8%(9/11),5个骨肉瘤细胞株均表达MICA mRNA;MICA蛋白在骨肉瘤组织和正常骨组织的表达率分别为51.6%(34/66)和0%(0/6);骨肉瘤细胞株Saos-2、MG63和HOS阳性表达膜表面MICA蛋白,而细胞株U2OS和OS732只有少量细胞呈阳性表达。【结论】MICA mRNA和蛋白广泛表达于骨肉瘤组织和细胞株。
