群体反应性抗体与HLA分型技术在肾移植上的联合应用

目的 总结群体反应性抗体（PRA）与HLA分型在肾移植联合应用上的意义。方法 PRA应用ELISA方法,HLA－Ⅰ、ⅡAg应用单克隆抗体分型技术,对20例肾移植病人手术前的PRA和HLA分型结果进行分析。结果受者PRA20％以下者12例,其余8例为20％－50％,供受者HLA－A、B、DR完全符合交叉反应组（CREG）为13例。其中A、B、DR分别有一相同抗原的为6例。术后11例发生急性排斥,经冲击治疗9例逆转,2例肾脏摘除91例PRA为20％,1例PRA为40％）。18例肾功能恢复。结论 PRA与HLA配型联合应用,可提高肾移植效果。     

PRA配型技术在肾移植中的应用研究

目的探讨群体反应性抗体 (PRA)配型新技术对肾移植近远期的效果。方法 85 4例患者肾移植前运用PRA新技术进行组织配型 ,并行血浆置换 ,未采用PRA组织配型的 42 3例作为对照 ,观察肾移植术后免疫指标变化、近期 (AR)发生率以及对长期存活的影响。结果未采用PRA组织配型组发生超急性排斥反应 (HR) 9例 (2 1% )、急性排斥反应 198例 (47% ) ;1年人/肾存活率 86 7% /76 3%、3年人 /肾存活率 72 5 % /6 7 9%、5年人 /肾存活率 6 9 5 % /4 9 3%。采用PRA配型新技术共 85 4例 ,肾移植术后未发生超急性排斥反应 ,发生急性排斥反应 16 2例 (19 0 % ) ,1年人肾存活率达 97 3% /95 0 %、3年人肾存活率 92 0 % /84 2 %、5年人 /肾存活率 87 0 % /81 6 %。结论PRA阴性配型可杜绝超急性排斥反应发生 ,降低急性排斥反应发生率 ,提高人 /肾长期存活率。     

The importance of PRA detection and PRA clearance in cadaveric renal transplantation

Though complement-dependent cytototicity (CDC)asSay is widely adopted as a standald histocompatibilitytest before haplantalon in most of the transplantationcenters in our countw, it yields unacceptably high ~ ofhypemeute rejechon (HR) Of the allograft because of itslow sensihvity[']. In some cases, even the new powerful~nosuPPressants fail to suPPress the disastIDus edejection (GR). In this stUdy, we examined the validity.Of Panel reactive antibodies (PRA) measmnt as a pretranSPlant h…     

亲属肾移植患者术后肾功能与HLA配型和PRA相关性研究

目的研究活体亲属肾移植受者人类白细胞抗原(human leukocyte antigen,HLA)配合率、群体反应性抗体(panelreactive antibody,PRA)的产生和肾功能变化对移植肾存活的影响。方法将336例活体亲属肾移植患者分为5组:①父母给子女供肾组(118例)。②子女给父母供肾组(12例)。③兄弟姐妹间供肾组(107例)。④其他亲属供肾组(92例)。⑤夫妻间供肾组(7例)。进行HLA供受者配合率分析,并于肾移植术后1-4年追踪肾功能变化和PRA产生的情况。HLA分型检测采用美国One lanmbda公司提供的HLA-PCR-SSP分型试剂盒。PRA采用美国莱姆德公司和美国GTI公司提供的酶联免疫吸附试剂盒检测。结果第1组的118例肾移植供受者HLA-A、B、DR、DQ抗原单体型半相合,其中有22例抗原配合率高于单体型半相合。肾移植术后有36例患者出现肾功能下降,其中8例为PRA阳性。第2组的12例肾移植供受者HLA-A、B、DR、DQ抗原单体型半相合,其中有7例HLA抗原配合率高于单体型半相合。肾移植术后1例患者肾功能下降,且为再次肾移植患者。第3组的107例兄弟姐妹间HLA-A、B、DR、DQ配型完全相配合有18例,73例为单体型半相合或大于单体型半相合,其余为低于单体型半相合或不相合。肾移植术后中有13例患者肾功能下降,其中3例PRA阳性。第4组的92例其他亲属移植的供受者间HLA-A、B、DR、DQ等于或大于单体型半相合的有24例,完全不相合的有9例,虽然HLA抗原配合率大于4个抗原,但并不是单体型半相合抗原的有8例,等于或小于3个抗原配合的有51例。肾移植术后有11例患者肾功能下降,其中6例患者PRA阳性。第5组的7例夫妻间肾移植患者,HLA配合率均≤3个抗原。肾移植术后有2例患者肾功能下降,且为PRA阳性。结论父母、子女及兄弟姊妹等直系亲属肾移植供受者中HLA配合率高于其他亲属间移植供受者,但兄弟姊妹间HLA配型完全相同的则较低。HLA配型与近期移植肾存活无关,而与供者的年龄有关。良好的HLA配型与肾移植术后PRA生成的概率低有关。     

肾移植致敏受者的配型研究及应用

目的:探讨人类白细胞抗原(HLA)配型在肾移植致敏受者中的应用及意义。方法:对32例PRA阳性的肾移植受者采用HLA交叉反应组(CREGs)配型原则选择供者,观察其供受者相配情况及移植效果。结果:按交叉反应组配型原则,供受者HLACREGS+DR0,1,2和3错配(MM)分别为8例(25％)、10例(31.25％)、12例(37.5％)和2例(6.25％),无4～6错配;而按传统的HLA-A,B,DR抗原配型原则,其0,1,2,3和4MM分别为2例(6.25％)、3例(9.38％)、10例(31.25％)、9例(28.12％)和8例(25％),其中0～1错配仅有5例。肾移植术后仅有9例发生急性排斥反应,经OKT_3治疗逆转,所有受者术后肾功能均恢复正常。结论:良好的HLA CREGs配型,可以显著提高供受者相配率,对减少致敏受者的排斥反应,提高移植肾存活率具有重要的意义。     

Role of Molecular Typing in Live Related Donor Renal Transplantation

In renal transplantation, a good HLA-DR match is associated with higher success rate of graft outcome. It is particularly so In high risk recipients. Serological HLA-DR typing is not always easy due to a number of technical problems. In view of this, a comparison of serological and molecular typing was done in our institutions. A total of 64 live related donor patients of renal transplantation were studied. Serological typing was done by conventional methods. Molecular HLA class II typing was done by polymerase chain reaction (PCR) based sequence specific oligonucleotide probe (SSOP) hybridization technique. An overall discrepancy of 19.5% was observed in the DR typing obtained by serology and PCR-SSOP of all the recipients and donors. 14.5% of cases showed discrepancy in the results of only one DR antigen. Serological typing failure was seen in 10.9% of total cases. In 19.5% cases, only one DR antigen was assigned by PCR-SSOP as compared to two antigens by serological methods. Maximum number of discrepancies were seen in DR 2 antigens. There was no appreciable difference of graft survival shown in the patients typed by both methods. However, higher incidence of acute graft rejection episodes were seen in patients with 1 antigen mismatch as compared to zero mismatch. It is concluded that HLA-DR typing should be carried out by molecular methods as these have been found to be more specific and accurate.Key Words: HLA-DR, Molecular typing     

Impact of human leukocyte antigen matching and recipients′ panel reactive antibodies on two-year outcome in presensitized renal allograft recipients

Background Renal transplantation in sensitized candidates remains a highly significant challenge worldwide. The production of panel reactive antibody （PRA） against human leukocyte antigen （HLA） is a major risk factor in presensitized recipients. The aim of this study was to evaluate the impact of HLA matching and recipients＇ PRA on two-year outcome in presensitized renal allograft recipients.
Methods We determined the percentage of panel reactivity and specificity of anti-HLA immunoglobulin （Ig） G antibodies in 73 presensitized renal allograft recipients compared with 81 unsensitized recipients （control group）. HLA genotyping of both recipients and corresponding donors was performed by PCR with sequence-specific primers （PCR-SSP）. We analyzed the factors influencing the early graft outcome （two-year rejection rates and survival rates of the grafts）, including HLA mismatching, class and degree of panel reactivity, and target antigen of donors.
Results Presensitized recipients had a worse two-year outcome than unsensitized recipients （P=0.019 for rejection rate, P=0.01 for survival rate）. The difference in number of HLA-mismatched alleles with either 6-antigen matching （Ag M） standard or amino acid residue matching （Res M） standard was not significant between the rejection and non-rejection groups of presensitized recipients or between the graft survival group and graft loss group. Compared with the control group, recipients with both PRA-I and PRA-II antibodies had a significantly worse two-year outcome （P=0.001 for rejection rate, P=0.002 for survival rate）. The two-year outcomes of the peak PRA 〉50% group and its subgroup, at-transplant PRA 〉50% group, were significantly worse compared with the control group （P=0.025 and P=0.001 for rejection rate, P=0.043 and P=0.024 for survival rate）. The rejection rates of the at-transplant target antigen positive group and its subgroup, HLA-I target antigen positive group, were significantly higher than the control group （P=0.001 and P=-0.001）, target antigen negative group （P=0.003 and P=0.001）, and peak target antigen positive with negative at-transplant target antigen group （P=0.024 and ,0=-0.002）. Two-year graft survival rates of the target antigen positive group and HLA-I target antigen positive group were significantly lower than the control group （P=0.012 and ,P=0.001）. The two-year outcome of target antigen unknown group was similar to that of the target antigen positive group. Presensitized recipients with pre-transplant plasmapheresis or immunoadsorption （PRA prepared group） had a better but non-significant two-year outcome than the control group. However, the PRA unprepared presensitized recipients were different to the control group （P=-0.004 for rejection rate and P=-0.005 for survival rate）. Hyperacute rejection （HR） occurred in three recipients with positive HLA-I target antigen and without mismatch according to Res M and in one case with positive PRA-II （for an unknown target antigen）. No HR occurred in eight cases with positive HLA-II target antigens.
Conclusions Pre-transplant PRA preparations might improve the access of presensitized patients to renal donors. Avoiding antigen-positive donors remains a fundamental measure in preventing HR and early rejections.     

良好的HLA配型可改善PRA高敏受者的移植效果

报告5例淋巴细胞群体反应抗体（PRA）为48％～76％，采用理想的HLA配型，供受者间HLA－A、B、DR位点在3～5个抗原相合情况下进行移植，术后3例发生急性排斥反应，经用甲基强的松龙及OKT3冲击治疗后排斥逆转。5例全部成功，术后均获得随访，人／肾存活9个月～2年4例，3年半1例。讨论了PRA高敏感对移植物的影响，良好的HLA配型对移植效果的影响。     

发作性睡病患者的人类白细胞抗原易感基因研究

目的　探讨人类白细胞抗原 (HLA)Ⅱ抗原基因在发作性睡病患者发病中的作用。方法　 31例患者经系统的病史询问、查体及头颅CT ,MRI检查排除了神经系统的其他疾患。均经多次小睡潜伏时间试验 (MSLT)测试及应用血清学方法进行HLADR2 测定 ;2 1例经特异性引物体外基因扩增 (PCR SSP)方法测定HLADR及HLADQ基因型。结果　 31例患者均符合发作性睡病的诊断。均有嗜睡、发作性猝倒 ,14例诉睡瘫 ,19例有入睡幻觉。MSLT试验示平均睡眠潜伏期为 2 1min± 1 3min ( 0 5～ 6min)。在 5次小睡中 ,30例患者出现异常的快动眼 (REM)睡眠 ,且均大于 2次 ,平均为4 2次± 1次 ( 2～ 5次 ) ,异常REM睡眠的潜伏期为 4 0min± 1 8min ( 0 8～ 7 9min)。在 31例患者中 ,HLADR2 阳性率为 96 8%( 30 / 31)。PCR SSP测定发现 2 0例HLADR2 阳性患者的亚型均为HLADRB 15 ,大部分是HLADRB1 15 0 1,但有 2例为HLADRB1 15 0 2。HLADQB1 0 6 0 2的阳性率为86 %( 18/ 2 1)。结论　HLADR2 及HLADQw6是发作性睡病的可能易感基因 ,但不同于文献报道东方人发作性睡病HLADR2 HLADQw6 10 0 %阳性的报道。     

HLA-DR位点基因分型在肾移植配型中的应用研究

应用聚合酶链式反应 序列特异性引物（ＰＣＲ ＳＳＰ）技术，对３３１例肾移植供受者进行ＨＬＡ ＤＲ位点基因分型，其中１０９例同时与血清学分型进行对比研究。显示血清学方法误差率为２９．７％；而ＰＣＲ ＳＳＰ法的ＤＲ位点分型，所有特异性均能显示，无假阴性及假阳性。随机抽取的２０例样本进行重复实验，重复性为１００％。分型结果经ＩＮＮＯ ＬＩＰＡＰＣＲ 反向ＳＳＯ实验确证，符合率为１００％。此方法耗时４ｈ，较血清学方法操作简便、快速、结果准确可靠，适合于临床器官移植ＨＬＡ分型。