HLA-G参与骨髓间充质干细胞的抑制作用机制探讨

目的：探讨人骨髓间充质干细胞（MSC）在活体肾移植供受者间的混合淋巴细胞培养（MLR）中的作用机制,证明人白细胞抗原（HLA）-Ⅰ类分子HLA-G参与了MSC的免疫调节作用。方法：从人骨髓中分离培养MSC,采用形态学观察及流式细胞术分析鉴定后,加入活体肾移植供受者间的MLR,观察MSC的HLA-G表达及对T细胞增殖的影响,淋巴细胞增殖实验采用MTT法。结果：MSC细胞表面和胞浆内均表达HLA-G,流式细胞术分析细胞表面HLA-G平均表达率为37.3%,胞浆内HLA-G平均表达率为65.1%,ELISA法检测细胞培养上清中可溶性HLA-G含量为18.9ng/mL。将MSC和培养上清液加入活体肾移植供受者间的MLR体系中,均使得T细胞抑制率增高;而加入HLA-G特异性抗体,MSC的抑制率降低。结论：MSC表面和胞浆内均有HLA-G表达,在其培养上清中检测到由MSC分泌的可溶性HLA-G5;MSC表达和分泌的HLA-G是其发挥抑制功能的关键因素之一。为MSC在预防器官移植术后排斥反应的临床应用提供了理论依据。     

组织相容性Ⅰ类相关链A位点基因多态性在预判肾移植物排斥中的意义

目的 研究组织相容性Ⅰ类相关链A位点(MICA)基因多态性和抗MICA特异性抗体在肾移植排斥反应发生中的意义.方法 采用免疫磁珠液相芯片技术对40例肾移植患者在移植术前和移植术后1个月、3个月、6个月、1年和2年动态检测抗MICA抗体的特异性和阳性分值的变化,同时采用SSOP方法分析16对肾移植供受者的MICA基因分型...     

肾移植受者生活质量和健康素养现状调查及影响因素分析

目的:调查肾移植受者生活质量(QOL)、健康素养(HL)现状,并分析其QOL、HL的影响因素。方法:选择我院2015年1月～2016年1月收治的369例肾移植受者为研究对象,采用自制问卷结合病历信息的方式收集入选患者的临床资料,分别采用健康状况调查简表(SF-36)、中文版成人快速健康素养评估量表(REALAM-T)对肾移植受者的QOL、HL的现状进行调查,并分析肾移植受者QOL、HL的影响因素。结果:369例肾移植受者QOL评分为(561.08±54.95)分,HL评分为(62.75±5.26)分,其中288例(78.05%)患者处于HL充足水平,56例(15.18%)患者处于HL临界水平,25例(6.78%)患者处于HL缺乏水平。单因素分析结果显示,不同婚姻状况、家庭人均月收入、文化程度、费用支付方式、移植肾来源、移植术后时间的肾移植受者QOL评分比较差异有统计学意义(P0.05);不同家庭人均月收入、文化程度、费用支付方式、移植肾来源、移植术后时间的肾移植受者HL评分比较差异有统计学意义(P0.05)。多元线性回归分析显示,家庭人均月收入、费用支付方式、移植肾来源、移植术后时间是肾移植受者QOL的影响因素(P0.05),文化程度、移植肾来源、移植术后时间是肾移植受者HL的影响因素(P0.05)。结论:肾移植受者QOL较差,HL整体不高,家庭人均月收入、费用支付方式、移植肾来源、移植术后时间是肾移植受者QOL的影响因素,文化程度、移植肾来源、移植术后时间是肾移植受者HL的影响因素,临床应根据以上因素采取针对性的措施,以提高肾移植受者的QOL和HL。     

环孢素A对大鼠心脏移植中血红素氧化酶-1表达的影响

目的　研究血红素氧化酶 1(HO 1)在大鼠同种异体心脏移植模型中的组织表达以及环孢素A (CsA)对该表达的影响。方法　心脏移植大鼠分为Ⅰ组 (对照组 )、Ⅱ组 (CsA 7 5mg/KgB W .)和Ⅲ组 (CsA 15mg/KgB .W .)。依据国际心肺移植组织 (ISHLT) 1990年心脏急性排异反应分级标准进行排异反应分析 ;免疫组化方法检测HO 1的表达。结果　心脏移植后 ,供者心脏和主动脉标本中 ,1和 3天时出现轻度炎性浸润 ,排异反应为 1A或 1B级 ;11天和 12天时出现弥漫性炎性浸润和心肌坏死 ,排斥级别达 3B或 4级。CsA治疗可使心脏移植排异反应降低 1～ 2 5级。移植后HO 1广泛表达于受体各类组织中 ,其中尤其以肝、肾、脾等器官表达增强比较明显 ,供心心肌细胞、血管内皮细胞和部分浸润淋巴细胞的HO 1表达均显著升高 ,CsA在一定时间内可明显增强供者心脏及主动脉的HO 1表达 ,减低移植排斥反应 ,从而延长移植心脏存活时间。结论　HO 1在心脏移植后受者体内表达广泛增强 ,CsA治疗可增强HO 1表达 ,这可能与其减弱移植排斥反应机制相关。     

他克莫司和环孢素A对肾移植术后治疗的Meta分析

目的:评价肾移植术后他克莫司(FK506)与环孢素A(Cs A)为基础治疗的有效性与安全性。方法:通过电子检索中国知网CNKI、万方、维普等数据库近五年相关文献,对纳入的文献进行数据提取后进行质量评价,并采用Rev Man5.2软件进行分析。结论:他克莫司与环孢素A可降低急性排斥反应和高血脂症发生率,同时对肝起到一定的保护作用,但是会引起血糖异常,对于糖尿病患者建议使用环孢素A。     

肾移植排斥患者肠道菌群变化的研究

摘要：目的 研究肾移植术后排斥患者肠道菌群变化的特点。方法 比较肾移植术后排斥患者及未排斥患者肠道菌群的变化,分析肠道菌群与肾移植排斥反应的相关性;以肾功能相关指标：肌酐（Scr）、尿素（BUN）、胱抑素C（Cys-C）、移植肾超声、移植肾穿刺活检评估肾脏移植患者排斥反应状态;检测患者淋巴细胞对供者骨髓源性树突状细胞（DC）刺激的反应,评价肾移植术后患者的免疫反应功能。结果 肾移植术后,排斥患者粪便中类杆菌含量[（5.53±0.74）CFU/g]明显少于未排斥患者[（6.84±1.66）CFU/g]（P<0.05）;双歧杆菌数量、肠球菌数量与肾移植患者尿素水平相关（r=－0.482,P=0.027;r=0.439,P=0.046）;类杆菌数量与肾移植患者尿素、肌酐、胱抑素C相关（r=－0.457,P=0.037;r=－0.515,P=0.014;r=－0.463,P=0.035）。排斥组患者的淋巴细胞对供者骨髓源性树突状细胞刺激的强度显著高于未排斥组（P＜0.05）。结论 肾移植术后排斥反应患者肠道菌群发生了显著改变,并与肾功能指标密切相关,此发现为合理应用抗排异药物、抗生素以及微生态制剂提供了理论依据,或可为减轻移植排斥提供新的思路和手段。     

HLA-G在肿瘤组织中表达情况研究进展

人类白细胞抗原G(human leukocyte antigen,HLA-G)属于非经典HLA-I类分子,在多种肿瘤细胞上均有表达。从结构上可以将HLA-G分为7种亚型:膜结合型HLA-G1-HLA-G4和可溶型HLA-G5-HLA-G7。研究表明,HLA-G1和HLA-G5具有明确的生物学活性也是研究较为深入的两种亚型,他们可以与T淋巴细胞、B淋巴细胞和NK细胞表面的ILT2/CD85j/LILRB1,ILT4/CD85d/LILRB2,KIR2DL4/CD158d受体结合而发挥免疫抑制功能。目前,HLA-G分子可以在肝癌、肾癌、肺癌、胃癌、食道癌、鼻咽癌、卵巢癌、乳腺癌、宫颈癌、直肠癌和血液肿瘤中表达。本文从HLA-G分子的结构和功能出发,综述了HLA-G分子在上述肿瘤中表达的情况,旨在分析HLA-G在各种肿瘤组织中表达的特点以及临床意义,为临床早期诊断和治疗肿瘤提供参考。     

大鼠肾移植慢性排斥反应模型的建立

目的建立稳定而可靠的大鼠肾移植慢性排斥反应模型。方法选用30只Wistar大鼠为供体,30只SD大鼠为受体。取供体左肾,采用HC-A离体肾保存液原位灌注,将供肾动、静脉分别与受体腹主动脉、下腔静脉行端侧吻合,以输尿管膀胱植入法行尿路重建,建立大鼠同种异体肾移植模型。分别于术后3、6、9周取移植肾观察大体和组织形态学变化,观察术后并发症及排斥反应情况。结果移植肾脏大体和组织形态学呈渐进性变化,至术后9周可出现明显的慢性排斥反应病理改变。移植肾脏可顺利存活,部分出现肾积水并发症,但不影响排斥反应病理变化。结论本方法可建立稳定、可靠的肾移植慢性排斥反应模型,是研究慢性排斥反应的理想模型。     

人白细胞抗原G在结直肠癌中的表达及意义

目的：在蛋白水平检测人白细胞抗原G（HLA-G）在结直肠癌组织中的表达,探讨HLA-G分子在结直肠癌不同分级和不同分期中的表达差异及在肿瘤逃逸中作用。方法：用免疫组织化学法检测结直肠癌和癌旁正常结直肠组织HLA-G的表达情况,用SPSS13.0软件、Kruskal-Wallis test进行分析。结果：HLA-G分子在结直肠癌组织中的表达率为42.3%（41/97）,在癌旁正常结直肠组织的表达率为0（0/20）;HLA-G分子表达与结直肠癌临床TNM分期（P〈0.05）和组织学分级（P〈0.01）相关。结论：HLA-G分子在结直肠癌组织中表达上调,且与结直肠癌的侵袭性生长密切相关;HLA-G分子可能下调宿主对肿瘤细胞的免疫应答反应,使肿瘤细胞逃避机体的免疫监视。     

Frequency of HLA-G exon 8 polymorphisms and kidney allograft outcome in Iranian population

The 14-bp polymorphism in exon 8 of the HLA-G gene is associated with HLA-G mRNA stability and the patterns of alternative isoform splicing and may influence the functionality of the HLA-G molecule. HLA-G expression was related to allograft acceptance and fewer episodes of acute rejection during heart, kidney and liver–kidney transplantation. In order to determine a possible correlation between the 14-bp insertion/deletion polymorphism and kidney allograft outcome in our population, genomic DNA was isolated from 144 patients who had received isolated kidney allografts. The recipients was divided into two groups, grafts presenting features of rejection group and a non-rejection group, and compared them with a control group of 100 healthy subjects. There was no significant difference in allelic frequencies of 14-bp insertion/deletion polymorphism between normal controls and kidney transplant patients. No significant difference was found between the RG and the NRG regarding the 14-bp genotypes and alleles. Therefore, additional studies with more sample size from other populations with analysis of other HLA-G polymorphisms are necessary to define this polymorphism as a valuable clinical marker.     

同种异体血管移植后肿瘤坏死因子表达的变化

目的探讨同种异体血管移植后移植血管和心肌肿瘤坏死因子(tumornecrosisfactor,TNF)表达的变化。方法取成年、雄性日本大白兔作为血管受者,新西兰大白兔为血管供者,进行颈动脉血管移植,通过免疫组织化学方法检测移植血管和心肌TNF表达的变化。结果自体原位移植组和液氮冷冻组TNF表达率低于新鲜移植组和抗生素处理组,新鲜移植组TNF表达率最高,与其它各组比较有显著性差异,P<0·05。血管移植后,心肌组织切片结果与血管移植组有类似结果:新鲜移植组心肌的TNF表达最高,与原位移植组和液氮冷冻组比较有显著性差异,P<0·05,而与抗生素处理组比较无显著差异性。结论液氮冷冻处理可降低移植血管和心肌TNF的产生;缺血再灌注损伤和/或免疫排斥反应能激发移植血管和全身各组织器官TNF的产生,从而造成移植血管的病理损害。     

Involvement of interaction between viral VP466 and host tropomyosin proteins in virus infection in shrimp

The C3 component of complement has different roles in kidney disease and its local production in donor kidney may affect allograft function and rejection after organ transplantation. A single base substitution in c3 gene (rs2230199), defines two common allelic variants with different mobility on gel electrophoresis: S (Slow) and F (Fast). In order to evaluate the effect of this polymorphism on acute renal allograft rejection, one hundred samples of donor and recipients were collected and genotyping was done by PCR-RFLP method. The allelic frequencies were: C3S=0.791, C3F=0.209. There was no significant association between recipient's genotype and acute rejection (p value<0.05). No correlation between donor genotype and acute rejection was also present. Patients were divided into four groups, according to the recipient and donor genotypes: SS recipients and FS or FF donor, SS recipient and donor, FF or FS recipient and SS donor and FS or FF recipient and donor. There was no significant difference in rate of acute rejection between groups. Although the results didn't find any association between C3 complement polymorphisms and acute allograft rejection, there was no acute rejection in FS or FF donors and SS recipient group.     

Assessment of different biomarkers provides valuable diagnostic standards in the evaluation of the risk of acute rejection

Acute rejection (AR) is a strong risk factor for chronic rejection in renal transplant recipients. Accurate and timely diagnosis of AR episodes is very important for disease control and prognosis. Therefore, objectively evaluated the immune status of patients is essential in the field of post-transplantation treatment. This longitudinal study investigated the usefulness of five biomarkers, human leukocyte antigen (HLA)-G5 and sCD30 level in sera, intracellular adenosine triphosphate (iATP) release level of CD4(+) T cells, and granzyme B/perforin expression in peripheral blood mononuclear cells (PBMCs) and biopsies, to detect AR and the resolution of biomarkers in a total of 84 cases of renal transplantation. The data demonstrated that recipients with clinical or biopsy proven rejection significantly increased iATP release level of CD4(+) T cells, and elevated sCD30 but lowered HLA-G5 level in sera compared with individuals with stable graft function. Expression levels of granzyme B and perforin were also elevated in PBMCs and graft biopsies of AR patients. Taken together, we identified that upregulation of sCD30, iATP, granzyme B, perforin, and downregulation of HLA-G5 could provide valuable diagnostic standards to identify those recipients in the risk of AR. And iATP may be a better biomarker than others for predicting the graft rejection episode.     

Polymorphisms in CTLA4 Influence Incidence of Drug-Induced Liver Injury after Renal Transplantation in Chinese Recipients

Genetic polymorphisms in cytotoxic T lymphocyte-associated antigen 4 (CTLA4) play an influential role in graft rejection and the long-term clinical outcome of organ transplantation. We investigated the association of 5 CTLA4 single-nucleotide polymorphisms (SNPs) (rs733618 C/T, rs4553808 A/G, rs5742909 C/T, rs231775 A/G, and rs3087243 G/A) with drug-induced liver injury (DILI) in Chinese renal transplantation (RT) recipients. Each recipient underwent a 24-month follow-up observation for drug-induced liver damage. The CTLA4 SNPs were genotyped in 864 renal transplantation recipients. A significant association was found between the rs231775 genotype and an early onset of DILI in the recipients. Multivariate analyses revealed that a risk factor, recipient rs231775 genotype (p = 0.040), was associated with DILI. Five haplotypes were estimated for 4 SNPs (excluding rs733618); the frequency of haplotype ACGG was significantly higher in the DILI group (68.9%) than in the non-DILI group (61.1%) (p = 0.041). In conclusion, CTLA4 haplotype ACGG was partially associated with the development of DILI in Chinese kidney transplant recipients. The rs231775 GG genotype may be a risk factor for immunosuppressive drug-induced liver damage.     

Altered Osteocyte-Specific Protein Expression in Bone after Childhood Solid Organ Transplantation

Background

Bone fragility is common post solid organ transplantation but little is known about bone pathology on a tissue level. Abnormal osteocytic protein expression has been linked to compromised bone health in chronic kidney disease (CKD) and immunosuppressant medications may impact osteocyte function.

Methods

Transiliac bone biopsies were obtained from 22 pediatric solid organ allograft recipients (average age 15.6 years) an average of 6.3 ± 1.2 years after transplantation and from 12 pediatric pre-dialysis CKD patients (average age 13.2 years). Histomorphometry and immunohistochemistry for FGF23, DMP1, sclerostin, and osteopontin were performed on all biopsies.

Results

FGF23 and sclerostin were increased in transplant recipients relative to non-transplant CKD, regardless of the type of allograft received and despite, in the case of liver and heart recipients, a higher GFR. Bone DMP1 expression was higher in liver or heart than in kidney recipients, concomitant with higher serum phosphate values. Osteopontin expression was higher in CKD than in transplant recipients (p<0.01). Bone FGF23 and sclerostin correlated directly (r = 0.38, p<0.05); bone FGF23 expression and osteoid thickness correlated inversely (r = - 0.46, p<0.01).

Conclusions

Solid-organ transplantation is associated with increased FGF23 and sclerostin expression. The contribution of these findings to compromised bone health post transplantation warrants further evaluation.     

Treatment with cyclosporin and risks of graft rejection in male kidney and heart transplant recipients with non-O blood.

In a consecutive series of 146 kidney transplant recipients treated with cyclosporin A a strong correlation between matching for the HLA-A, HLA-B, and HLA-DR loci specificities and outcome of the grafts was observed in male recipients with non-O blood groups. Such a beneficial effect of matching was not found in female patients or male patients with blood group O. In these patients survival of the grafts at one year was good irrespective of the number of HLA-A, B, and DR mismatches. Also in 47 male heart transplant recipients immune responsiveness against mismatched HLA antigens was related to blood group. A significantly higher incidence of rejection episodes was observed in male patients with non-O blood groups (n = 32) than in those with blood group O (n = 15). Matching for HLA-DR reduced the number of acute rejection episodes in male patients with non-O blood. These findings may help explain the controversial reports about the importance of HLA matching in organ transplantation. Furthermore, as most candidates for heart transplantation are male and not of blood group O, the higher incidence of graft rejection in these patients underscores the need for an exchange strategy of donor hearts.     

PDIA3 mRNA expression and IL-2, IL-4, IL-6, and CRP levels of acute kidney allograft rejection in rat

Kidney transplantation to treat end-stage renal disease has evolved rapidly from the first successful transplantations to the current widespread use of grafts from both cadaveric and living donors. But acute rejection is still a strong risk factor for chronic rejection in recipients of renal grafts. To investigate possible mechanisms, we describe a comparison between differentially proteins expression and immune markers profile (IL-2, IL-4, IL-6, and CRP) of acute rejection and the controls. Through quantitative real-time RT-PCR confirmation, PDIA3 mRNA and protein expression levels in serum and transplanted kidney in experiment group was significantly (P < 0.05) higher than that in control group. Immunity analysis showed that plasma IL-2, IL-4, IL-6, and CRP levels were higher in experimental rats than those in control rats. Our data thus indicate that PDIA3 might be potentially involve into the occurence and development of acute rejection response in renal transplantation and increased plasma IL-2, IL-4, IL-6, and CRP levels play an important role to prevent acute kidney allograft rejection in rats.     

TLR4在急性肝衰竭模型大鼠中的动态变化及意义

目的 探讨TLR4在D-Gal/LPS诱导的大鼠急性肝衰竭中的表达变化及其作用。方法 65只SD雄性大鼠随机分为正常对照组（n=5）、D-Gal/LPS模型组（n=25）、PDTC干预组（n=25）,于不同时间点检测肝功能,免疫组化观察肝组织TLR4的表达,TUNEL法观察肝脏细胞凋亡。结果 模型组大鼠4h开始ALT、AST明显升高,8h达峰值,4-24h ALT、AST明显高于正常对照组（P〈0.05）。模型组各时间点TLR4表达明显高于正常对照组（P〈0.05）。模型组凋亡的肝脏细胞数随时间的延长逐渐增多,均高于正常对照组（P〈0.05）。PDTC干预后ALT、AST水平,TLR4的表达及凋亡的肝脏细胞数低于模型组（P〈0.05）。结论 急性肝衰竭模型鼠TLR4表达增强,PDTC干预可下调其表达,提示TLR4在急性肝衰竭的发生发展中可能具有一定作用。