HLA-DQA反相杂交法分型与多聚酶链反应－单链构象多态性配型的比较

为了验证多聚酶链反应－单链构象多态性方法在骨髓移植配型中的可靠性和准确性，采用掺入生物素的多聚酶链反应对ＨＬＡ－ＤＱＡ位点基因进行了特异性扩增。将所选区分ＨＬＡ－ＤＱＡ位点等位基因的特异性探针点于尼龙膜上，用标记的扩增产物与膜上探针杂交，并用碱性磷酸酶显色法检测杂交信号，确定待测标本的基因型。对２０对异基因骨髓移植的供、受者进行ＨＬＡ－ＤＱＡ位点的基因分型，并与多聚酶链反应－单链构象多态性配型作比较。结果表明，２０对供、受者的多聚酶链反应－单链构象多态性配型结果与该反相杂交法基因分型结果一致。多聚酶链反应－单链构象多态性方法是一种骨髓移植的供、受者配型，快速可靠的方法     

HLA-D基因配型的单链构型多态性银染分析

用PCR/单链构型多态性(SSCP)/银染方法做HLA-D基因配型。取供、受者DNA,对HLA-DQA、DRB、DQB及DPB位点作PCR(第2外显子多态部分),然后对产物进行SSCP,电泳条带经银染后分析结果,以确定供、受者HLA-D基因型是否一致,银染较之溴化乙锭染色灵敏度高25倍左右,为骨髓移植前进行HLA-DQA、DRB、DQB及DPB基因配型提供了较为灵敏的非放射性的手段。该方法亦可应用于其它单链构型多态性分析。     

96例血小板基因配型患者的HLA-Ⅰ类抗体特性分析及抗原免疫原性估算

目的 分析血小板基因配型患者的HLA-Ⅰ抗体特异性,并估算HLA抗原免疫原性。方法 标本来自医院送检本中心申请血小板基因配型的血小板输注无效(PTR)患者,共计96份。采用Luminex技术对患者标本进行HLA-Ⅰ抗体特异性检测,并根据MFI分析抗体表达水平。人群HLA抗原错配率、相对免疫原性根据HLA-A、-B位点等位基因频率分布和抗体检出频次分别进行估算。结果 96份患者标本均检出HLA-Ⅰ类抗体,但抗体种类分布较广。HLA-A、-B、-C磁珠包被抗原(共计97个)对应抗体的平均阳性检出率分别为0.38、0.47、0.28。在浙江人群中HLA-A和-B系统中,抗原频率较高的分别为HLA-A2、A11、A24和HLA-B60、B46、B58,其相对免疫原性分别为0.403、0.283、0.342和0.100、0.067、0.178。结论 PTR患者存在不同种类特性的HLA-Ⅰ抗体,证实不同HLA-A、-B抗原的相对免疫原性存在差异。     

HLA-Ⅰ类分子与肿瘤免疫

人MHCI类基因分两类:经典的Ⅰa基因(包括HLA-A、HLA-B、HLA-C)和非经典的Ⅰb基因(主要包括HLA-E与HLA-G、HLA-F).经典的HLA-Ⅰ类基因集中在远离着丝点的一端,包括B、C、A三个座位,到1996年WHO命名委员会公布的HLAI类基因中,已发现HLA-A位点有61个复等位基因,B位点和C位点各有136个和37个复等位基因,其产物称为HLA分子,经典的HLAI类抗原(HLA-A、B、C系列)广泛分布于人体各种组织的有核细胞表面,各种组织细胞表达HLA-Ⅰ类抗原的数量不同,以外周血白细胞和脾淋巴结,胸腺细胞的含量最丰富,其次为肺、肝、肾、皮肤、主动脉和肌肉.     

HLA-DRB PCR-单链构象多态性分析

采用聚合酶链技术(PCR)扩增10例HLA-DR血清学定型者的DRB基因,然后作单链构象多态性(SSCP)分析,发现不同基因型的DNA单链区带数及电泳速度不同,提示本方法可用于器官移植时供、受者DRB基因配型及法医犯罪识别。     

用PCR-指纹图和PCR-单链构象多态性对骨髓移植供、受体进行HLA-D区配型

为探讨HLA血清学分型与DNA配型结果的相关性,本文建立了HLA-DQA、DQB、DPB、DRB位点聚合酶链反应-指纹图(PCR-F)、聚合酶链反应-单链构象多态性(PCR-SSCP)配型方法,并用PCR-F、PCR-SSCP对21对骨髓供、受者进行了基因配型。与血清学结果比较,其中10对血清学配型一致的供、受体,在DQA位点PCR-F、PCR-SSCP有8对相合,在DRB位点有7对相合。提示在配型中,应用PCR-F和PCR-SSCP可发现血清学不易检测出的基因亚型。     

骨髓移植供受者HLA－DRB的单链构型多态性配型

用ＰＣＲ和单链构型多态性（ＰＣＲ－ＳＳＣＰ）法检查供受者的ＨＬＡ－ＤＲＢ－１基因座的多态性的一致性，以此做ＤＲＢ的基因配型。１６例同胞供受者对的ＤＲＢ－１的ＰＣＲ－ＳＳＣＰ配型，有６对相合（６／１６），１０对不合（１０／１６）。此基因配型结果与其它配型结果比较，说明ＤＲＢ－１的ＰＣＲ－ＳＳＣＰ配型在区分同胞供受者对的Ⅱ类抗原多态性上有较高的灵敏度，同时表明此类配型在应用于骨髓移植前检查同胞供者的意义。     

HLA-A、B、DR血清学分型与基因分型的比较研究

目的比较研究HLA-Ⅰ、Ⅱ血清学分型与基因分型结果,分析HLA-A、B、DR血清学分型误定规律,提高移植配型的准确性。方法应用聚合酶链反应-序列特异性引物(PCR-SSP)技术,对240名骨髓资料库中已用血清学分型的自愿者进行HLA-A、B、DR基因分型,并对血清学分型与基因分型结果进行低分辨水平的比较研究。结果HLA-A特异性血清学分型错误率在纯合子与杂合子中分别占30.65%与11.52%,总错误率14.35%;HLA-B特异性血清学分型错误率在纯合子与杂合子中分别占42.22%与16.15%,总错误率18.85%;HLA-DR特异性血清学分型错误率在纯合子与杂合子中分别占37.50%与14.58%,总错误率18.63%。HLA-A特异性血清学分型假阴性16.55%,假阳性1.44%,错误指认特异性7.42%,HLA-B分别为20.32%,1.84%和13.56%,HLA-DR分别为13.33%,2.21%和10.05%。结论HLA-A、B、DR纯合子血清学分型错误率明显高于对应的杂合子分型,HLA-B,DR特异性血清学分型错误率显著高于HLA-A特异性;为了提高移植配型的准确性,骨髓资料库中自愿者HLA-A、B、DR位点须重新用基因分型方法分型。     

低分辨率和高分辨率HLA-Ⅱ类PCR-SSP基因分型在骨髓移植中的应用

为满足临床骨髓移植对HLA配型的要求,本研究采用先进、快速的DNA提取方法和低分辨率与高分辨率HLA-ⅡPCR-SSP分型方法,对150例临床骨髓移植供、受者进行HLA-Ⅱ类分型研究,并且对分型方法进行了改进,建立了随时间释放DNA聚合酶活性的分型方法。结果表明:DNA提取方法可以满足微量HLA-Ⅱ类DNA分型方法对DNA样本的要求,200μl全血DNA产量为4-12μg,A_(260)/A_(280)为1.7～1.9。低分辨率和高分辨率HLA-Ⅱ类PCR-SSP均具有良好稳定性、可靠性和准确性。低分辨率HLA-Ⅱ类PCR-SSP方法耗时1.5小时,适用于同胞兄弟姐妹间的骨髓移植配型。高分辨率HLA-Ⅱ类亚型PCR-SSP分型方法耗时2小时50分钟,适用于同胞兄弟姐妹间骨髓移植配型的特殊病例和无血缘关系骨髓移植供、受者间的配型。本研究的分型方法对于推动我国骨髓移植工作的开展和未来的发展均具有重要的意义。     

HLA-DQ与妊娠期糖尿病关联的研究进展

人类白细胞抗原(human leucocyte antigen，HLA)是人类主要组织相容性复合体的编码产物。HLA基因位于第6号染色体短臂的2区l带，长度约3500kb，共有数十个基因座，可分为三类，分别命名为HLA-Ⅰ、Ⅱ、Ⅲ类基因。HLA-A、B、C为经典的HLA-Ⅰ类基因，而经典的HLA-Ⅱ类基因一般指HLA-DR、DQ、DP，它们编码双肽链(a、     

HLA低分辨相合非亲缘造血干细胞供受者高分辨全匹配的概率分析

本研究旨在分析HLA-A、-B、-DRB1低分辨位点相合的非亲缘关系造血干细胞供受者HLA-A、-B、-C、-DRB1和-DQB1高分辨匹配程度。采用PCR-SBT进行HLA-A、-B、-C、-DRB1和-DQB1位点高分辨检测。结果表明：在HLA-A、-B、-DRB1低分辨位点相合的274对非亲缘关系供受者标本中：①患者总数为166例,供者为274例。97例（58.43%）患者为1个供者,47例（28.31%）患者为2个供者。②HLA-A、-B、-C、-DRB1、-DQB1座位10个位点高分辨配合情况为6/10相合32对（11.68%）,7/10相合为54对（19.71%）,8/10相合为62对（22.63%）,9/10相合为49对（17.88%）,10/10相合为48对（17.52%）。③供受者HLA-A、-B、-C、-DRB1、-DQB1座位相合程度不同,以HLA-C位点不相合程度最高。274例供者检出23个HLA-A、46个HLA-B、21个HLA-C、30个HLA-DRB1和17个HLA-DQB1等位基因。166例患者检出20个HLA-A、40个HLA-B、22个HLA-C、29个HLA-DRB1和16个HLA-DQB1等位基因。④274例供者检出313条单倍型,166例患者检出224条单倍型,两者间频率最高的单倍型为A＊02：07-B＊46：01-C＊01：02-DRB1＊09：01：02-DQB1＊03：03（5.63%和6.88%）。结论：在HLA-A、-B、-DRB1低分辨位点相合的基础上,非亲缘关系造血干细胞供受者HLA-A、-B、-C、-DRB1和-DQB1座位高分辨完全相合的概率较低。     

Donor-specific transfusions. Donor-recipient HLA compatibility, recipient HLA haplotype, and antibody production

HLA profiles of 25 donor-specific transfusion (DST) kidney donor-recipient pairs were analyzed for HLA antigen compatibility. Serum samples collected during and after DST were tested for cytotoxic antibodies against T and B lymphocytes of the donors and 30 normal individuals. Eleven recipients did not produce cytotoxic antibodies to the antigens of their DST donors, and eight produced cold and/or warm, broadly reactive B-cell antibodies. Six patients (24%) produced HLA-A, B, C, and/or DR antibodies. Three of these individuals produced antibodies after two immunizations, while others required three immunizations. Three of the 11 antibody nonproducers (17%) had not received previous transfusions, as compared to three of the eight antibody producers (43%). Comparison of HLA profiles revealed 22 percent of the HLA-A, B, DR identities between the transfusion donor and recipient in antibody nonproducers as compared to 9 percent of the HLA-A, B, DR identities in antibody producers. The HLA-A2, B40, DR4 haplotype and HLA-DRW6 antigen were more common among antibody producers than among nonproducers, who had an excess of the HLA-B8, DR3 haplotype. These results are consistent with the hypothesis that there may be high- and low-responder HLA haplotypes that control immunologic responsiveness to histocompatibility antigens.     

HLA-I类基因与白血病相关性研究

本研究目的是从常见类型白血病群体的HLA多态性分布特征探讨白血病与HLA-Ⅰ类基因的相关性。采用序列特异性引物扩增方法（SSP—PCR）检测了605例白血病患者（ALL189例、AML184例、CML232例）的HLA-A、B、C基因特异性，以900份脐血库资料作为正常群体对照。结果发现，在急性淋巴细胞性白血病（LL）患者中HLA—A＊26、A＊68、B＊56的基因频率分别为4．46％、2．65％和1．17％，明显高于对照组（2.31％、0．95％和0．22％）；Cw＊06基因频率（3．64％）明显低于对照组（11．65％）。在急性髓性白血病（AML）患者中HLA—A＊01、B＊37基因频率（9．41％、3．60％）明显高于正常对照（3．57％、1．75％），A＊33、B＊51基因频率（3．60％、4．73％）明显降低（对照：7．64％、7．93％）；在慢性髓性白血病（CML）患者中HLA-A＊32、B＊27、B＊44、B＊54、B＊55基因频率分别为2．18％、3．96％、5．06％、4．63％和2．84％，明显高于正常对照（0．84％、2．04％、3．07％、2．44％和1．29％）。结论：各类白血病分别与不同的HLA基因有明显的相关性，这对白血病发病机制的研究有重要意义。     

标准HLA-A、B、DR配型与氨基酸残基配型在肾移植术后早期急性排斥反应中的差异

目的　研究标准HLA -A、B、DR配型和氨基酸残基配型与肾移植术后早期急性排斥反应中的差异。方法　将 14 47例次肾移植受者 ,按HLA误配率 (MM )对标准配型和氨基酸残基配型 ,分别进行分组 ,统计各组术后 1～ 2个月内 ,急性排斥反应的发生率。结果　标准HLA配型 6误配组 ,术后早期急性排斥反应发生率为 2 1 .6% ;5误配组为 2 8. 9% ;4误配组为 19 .1% ;3误配组为 16. 5 % ;2误配组为 16 .3 % ;1和 0误配组未见术后早期急性排斥反应发生。而氨基酸残基配型 6误配组 ,术后早期急性排斥反应发生率为 2 8 .6% ;5误配组为 2 0 . 2 % ;4误配组为2 3 . 6% ;3误配组为 2 0 . 2 % ;2误配组为 2 0 . 4% ;1误配组为 19% ;0误配组为 16%。结论　标准HLA -A、B、DR配型比氨基酸残基配型在肾移植术后早期急性排斥反应中的作用更为明显。     

应用不同类型白血病患者群体HLA-A、B和DRB1遗传学数据评估HLA相合供者的概率

目的探讨不同类型白血病汉族患者群体找到HLA-A、B和DRB1低分辨全相合无关供者可能性。方法依据1201名急性淋巴细胞白血病(ALL)、1065名急性非淋巴细胞白血病(AML)和1432名慢性粒细胞白血病(CML)汉族患者群体HLA-A、B和DRB1低分辨分型结果,计算这3类白血病汉族患者群体的HLA-A、B和DRB1抗原频率,单体型频率和表型频率,联合应用中华骨髓库(CMDP)中健康汉族供者HLA表型频率数据,进一步推导出这3类白血病汉族患者群体找到至少1名HLA低分辨全相合供者概率及回归方程。结果ALL、AML、CML汉族患者群体表型种类数分别为416851、373903和464373种,其中31%的表型频率均>1/100万;ALL、AML、CML汉族患者群体在CMDP的汉族供者群体中找到至少1名HLA表型相同供者回归方程分别为:P=0.0824LgN-0.3667、P=0.0825LgN-0.3636和P=0.0829LgN-0.3644;计算出81.6%的ALL、81.9%的AML、82.4%的CML汉族患者要找到1名HLA低分辨表型相同的无关供者,CMDP中汉族有效供者群体数平均146.7万。结论应用不同类型白血病患者群体HLA-A、B和DRB1遗传学数据评估患者找到HLA相合供者概率较仅以健康供者群体HLA遗传学数据评估更为精确。为有效地提高患者的移植效果,需要征募更多的无关供者。     

The changing activities of a regional marrow donor program

BACKGROUND: A regional marrow donor program was established in 1982. Following the establishment of the National Marrow Donor Program (NMDP) in 1987, the activities of this regional program changed. STUDY DESIGN AND METHODS: To better understand the changes that occurred in the regional marrow donor program, its donor recruitment and marrow collection activities through 1991 were studied retrospectively. Data analyzed included the total number of potential donors, the number and types of potential donors recruited each year, the number of searches performed, the number of samples collected for HLA-DR typing and mixed lymphocyte culture testing, and the number of transplants in both programs from 1987 through 1991. Statistical analysis was performed by using chi-square. RESULTS: Initially, only persons who donated platelets by apheresis were enrolled into the program. In 1986, the regional program's first drive to recruit people who were not apheresis donors occurred. The number of such drives increased each year, and in 1991, 12 drives occurred, which resulted in the recruitment of 1313 potential marrow donors. From 1987 to 1991, the number of potential donors in the regional program grew from 3252 to 9146, but the proportion of apheresis donors in the program decreased. In 1987, 91.9 percent of marrow donors at the regional center had been apheresis donors, but in 1991, 41.7 percent had been apheresis donors. The number of marrows donated at the regional center increased from 11 in 1987 to 29 in 1989, but then fell to 24 per year in 1990 and 1991. The decrease in the number of donations at the regional program was due to the rapid growth in the NMDP file of potential marrow donors and the selection of donors whose HLA antigens were more compatible with those of the transplant recipients. In 1989, the regional program contained 4.6 percent of all HLA-A,B-typed and 11.2 percent of all HLA-A,B,DR-typed potential donors in the NMDP and collected 15.3 percent of all marrows. However, in 1991, the regional program contained 2.0 percent of HLA-A,B- typed donors and 4.1 percent of HLA-A,B,DR-typed donors and collected 5.3 percent of marrows. In 1987, 18 percent of the people who donated marrow at the regional center were phenotypically HLA-A,B,DR identical with the recipient, but in 1991, 92 percent of donor-recipient pairs were phenotypically HLA-A,B,DR identical. CONCLUSION: Recruitment activities became an increasingly larger part of the Regional Marrow Donor Program's activities. Increasing the size of the file of potential donors was necessary to maintain a constant number of donations. Persons who were not regular blood donors were an important part of the marrow donor program.     

Potential for transfusion-associated graft-versus-host disease due to apheresis platelets matched for HLA class I antigens

Transfusion-associated graft-versus-host disease (TA-GVHD) has been reported in immunocompetent recipients of nonirradiated cellular blood components from donors who are homozygous for an HLA haplotype shared with the patient. In these cases, donor lymphocytes have no antigens foreign to the recipient, and this similarity in HLA antigens appears important for the development of TA-GVHD. Experience with 65 patients receiving apheresis platelets matched for class I HLA antigens was reviewed to determine the incidence of such a transfusion among HLA- matched, unrelated donor-recipient pairs. In 5 percent of transfusions (31/673), the patient received lymphocytes from a donor exhibiting no antigens foreign to the recipient, but the patient had additional HLA-A or -B antigens not present on donor lymphocytes. Twenty-three percent (n = 15) of patients received at least one such transfusion. In addition, most patients were immunosuppressed as a result of their underlying disease or therapy, which may decrease the degree of antigen matching required to initiate TA-GVHD. Until the pathogenesis of this disease is better understood, it is recommended that the transfusion of an HLA-matched cellular blood component be considered a risk factor for the development of TA-GVHD regardless of the patient's immune status, and that all such blood components be irradiated.     

Role of Ia-Like Products of the Main Histocompatibility Complex in Conditioning Skin Allograft Survival in Man

This report correlates the survival time of 93 intrafamilial skin allografts performed under conditions of main histocompatibility complex (HLA) haploidentity with donor-recipient compatibility for products of the HLA-A, -B, -C, and -DR, as well as C3 proactivator, Glyoxalase I, and P loci located on the human 6th chromosome. Incompatibilities for HLA-A and -B (and to a lesser extent for HLA-C) and(or) for HLA-DR products exerted a strong influence upon the fate of skin allografts. When HLA-A and -B were considered alone, the most compatible group of grafts had a mean survival time of 15.8 d, as compared with 11.3 d for the most incompatible transplants. HLA-DR compatibility alone was associated with a mean survival time of 15.3 d, whereas HLA-DR-incompatible grafts had a mean survival time of 11.5 d. Incompatibilities for C3 proactivator, Glyoxalase I, and P did not have a significant effect upon graft survival.There was no evidence of an association between donor-recipient incompatibility at HLA-A, -B, or -C or at HLA-DR; such incompatibilities occurred independently of each other, in spite of the state of linkage disequilibrium known to exist between HLA-B and -DR. Incompatibilities for HLA-A, -B, and for HLA-DR exerted a potent additive effect upon graft survival. Skin grafts bearing one, two, or three incompatibilities had a mean survival time of 16.2, 13.7, and 10.7 d, respectively (P <0.0005).The results point to the important role played by the Ia-like products of the HLA complex (HLA-DR) in conditioning skin allograft survival in man. This consideration may be of direct relevance to the potential clinical usefulness of in vitro serological techniques for the detection of donor-recipient compatibility for HLA-DR.     

中华骨髓库造血干细胞捐献志愿者HLA基因分型中模棱两可结果解决方案的探讨

目的 探讨中华骨髓库造血干细胞捐献志愿者HLA基因分型中模棱两可结果的解决方案。方法 采用聚合酶链式反应(PCR)-测序分型技术(SBT)对20621名中华骨髓库供者的HLA-A、B、DRB1基因进行高分辨分型，并对其中的模棱两可分型结果进行精确分型。根据精确分型后的HLA等位基因频率计算模棱两可结果中真基因型的相...