HLA分型方法的研究进展

通过对人类白细胞抗原(human leucocyte antigen,HLA)结构和功能的研究,有助于认识其在免疫学、人类学等领域中的作用,揭示其在疾病发生、发展中的作用机理,而对HLA分型研究却是认识其作用的关键一步。随着PCR技术的广泛应用,HLA基因分型方法得到了迅速的发展。本文对HLA基因分型技术的进展情况进行综述。     

HLA多态性及HIV进化与疾病关联的研究进展

HIV-1的高度多样性除受病毒自身的复制特征影响外,HLA-I类分子介导CTL免疫反应的选择压力造成的突变对HIV-1多样性的产生有一定作用。近期许多研究显示HLA-I类等位基因、单体型和超型与HIV-1多样性及疾病进程的关系,HIV-1多样性和编码HLA-I类分子基因的多态性是平行的,本文就HLA多态性与HIV多样性及疾病关联的研究进展进行综述。     

国产及进口HLA—B27分型试剂盒的比较

人类白细胞抗原(HLA)是机体内有高度多态性的抗原系统,在器官移植、输血、人类学研究等领域广泛的应用,B_(27)是HLA—B位点上的一个重要的等位基因,与强直性脊椎炎、反应性关节炎、急性虹膜炎等疾病相关,临床上已开始将B_(27)分型用于这些疾病的辅助诊断.目前许多实验室均是采用补体依赖性微量淋巴细胞毒试验进行检测,本文用国产及进口HLA—B_(270分型试剂来检测HLA—B_(27)抗原进行了比较研究.1 材料与方法1.1 材料 国产试剂购自北京解放军307医院免疫室.进口试剂由美国莱姆德公司北京办事处提供.1.2 方法 国产试剂的实验方法为通用的美国NIH微量淋巴细胞毒试验,进口试剂的实验方法按美国莱姆德公司提供.1.3 血标本 30个标准细胞采自广州医学院HLA细胞库,其中有5个带有HLA—B_(27)抗原.1.4 统计方法 评价血清特异性强度用X~2和相关系数估计,血清反应强度用强度指数SI表示:SI=记分为8的反应数/记分为6、8反应系数用中国军事医学科学院计算中心编制的HLA软件进行处理数据.2 结果     

DNA序列分析鉴定HLA新等位基因B*4446

目的鉴定中国人群的HLA新等位基因。方法使用PCR-序列特异性引物及PCR-序列特异性寡核苷酸探针技术进行HLA分型。发现1个与HLA-B*44相关的未知基因,使用DNA序列分析技术鉴定并分析该基因与同源性最高的B*4409基因序列的差异。结果新基因第3外显子区域序列与所有已知的HLA-B等位基因序列均不相同,与同源性最高的HLA-B*4409基因序列相比有3个碱基发生替代。第538位碱基由G→C,第539位碱基A→T以及第540位碱基C→G,使编码产物180位氨基酸由天冬氨酸变成亮氨酸。结论发现一个新的HLA-B等位基因,于2005年9月由WHOHLA因子命名委员会正式命名为HLA-B*4446。     

HLA－B27的分型方法

本文综述了ＨＬＡ－Ｂ２７的分型方法以及各种方法的此缺点。至今建立的ＨＬＡ－Ｂ２７分型方法有补体依赖性微量淋巴细胞毒法、流式细胞术法、玫瑰花法、等电聚焦法、酶联免疫法和聚合酶链反应（ＰＣＲ）法。这些方法虽各有其特点，但ＰＣＲ法分型ＨＬＡ－Ｂ２７亚型将是未来的发展方向。     

我国中部地区HIV感染者HLA Ⅰ类等位基因多态性与病毒载量的相关性研究

目的 分析我国中部地区既往采浆HIV感染者HLA Ⅰ类等位基因多态性及其与病毒载量的相关性.方法 应用PCR-SSP(序列特异性引物)技术对106例HIV感染者进行HLA Ⅰ类基因分型,分析HLA Ⅰ类等位基因多态性和单元型与血浆病毒载量的相关性.利用ELISPOT技术检测Gag蛋白特异性的CTL反应,并分析其与血浆病毒载量的相关性.结果 携带HLA Ⅰ类等位基因纯合子的感染者具有较高的病毒载量(P=0.0098);HLA-A*30、-B*13、-Cw*06、-Cw*14等位基因及A*30/B*13/Cw*06单元型与低病毒载量相关(P=0.0004,0.0103,0.0058,0.0371和0.0006);携带HLA-A*30/B*13/Cw*06单元型的感染者p2p7p1p6蛋白特异性的CTL反应与病毒载量负相关;携带HLA-Cw*14的感染者p17蛋白特异性的CTL反应与病毒载量负相关.结论 我国中部地区既往采浆HIV感染人群HLA-A*30、-B*13、-Cw*06、-Cw*14等位基因及A*30/B*13/Cw*06单元型携带者具有较低的病毒载量,其保护性与Gag蛋白特异性的CTL反应有关.     

HLA多态性与HIV感染及AIDS发病相关性的研究

为研究人类白细胞抗原（ＨＬＡ）和人免疫缺陷病毒（ＨＩＶ）易感性的关系，本文分析比较了以下三组人群的ＨＬＡ表型频率和单倍型频率：①１７２例正常人；②１７例血清ＨＩＶ阴性高危人群；③１８０例血清ＨＩＶ阳性患者，其中２１例发展为艾滋病（ＡＩＤＳ），３７例６个月内ＣＤ４＋淋巴细胞降低至少２０％（ＣＤ４ｄｅｃｌｉｎｅ）。在１７２例对照和１８０例血清ＨＩＶ阳性受试者的比较中发现，其ＨＬＡ表型频率和单倍型频率没有显著差别，提示ＨＩＶ感染与ＨＬＡ无关联。然而，ＨＬＡ－Ｂ２１、ＨＬＡ－Ｂ８、ＨＬＡ－Ｂ３５表型及ＨＬＡ－Ａ１－Ｂ８、ＨＬＡ－Ｂ８－ＤＲ３、ＨＬＡ－Ａ１－Ｂ８－ＤＲ３单倍型与ＣＤ４阳性淋巴细胞下降，或与血清ＨＩＶ感染发展成ＡＩＤＳ病显著相关。１７例ＨＩＶ血清阴性高危人群中，无一携带单倍型ＨＬＡ－Ａ１－Ｂ８－ＤＲ３，提示该单倍型可能与ＨＩＶ感染的抗性相关。     

应用SBT直接测序分型法研究中国南方汉族人群HLA五个基因座单倍型

目的 研究中国南方汉族人群HLA-A、B、Cw、DRBl、DQBl等位基因多态性及单倍型的分布特征.方法 应用聚合酶链反应-直接测序分型(polymerase chain reaction sequence-based typing,PCR-SBT)法对186名中国南方汉族健康人群HLA-A、B、Cw、DRBl、DQBl进行基因分型.结果 检出的HLA-A、B、Cw、DRBl、DQBl等位基因分别有28、49、24、29、20种.经统计分析A*0207-B*4601(10.81%),A*3303-B*5801(6.14%),B*4601-DRBl*0901(6.22%),B*4001*DRBl*0901(3.78%),DRBl*0901-DQBl*0303(12.16%)和DRBl*1202-DQBl*0301(8.38%)单倍型呈强连锁不平衡单倍型(RLF≥0.5,X<'2>>3.84,P<0.05);A*0207-B*4601-Cw*0102(10.75%),A*3303-B*5801-Cw*0302(5.14%),A*0207-B*4601-DR*0901(5.07%),A*3303-B*5801-DRBl*0301(2.96%),A*0207-B*4601-Cw*0102-DRBl*0901-DQBl*0303(4.87%)和A*1101-B*1301-Cw*0304-DRBl*1501-DQBl*0601(2.43%)单倍型分别是中国南方汉族人群常见单倍型.结论 中国南方汉族人群HLA 5个基因座单倍型分布具有高度的遗传多态性且有其自身分布特点.本研究获得的较完整的HLA 5个基因座单倍型分布数据,将为人类学、HLA疾病相关性和器官移植等研究提供遗传学参考数据.     

歧视知觉与男同性恋艾滋病感染者创伤后应激障碍的关系:应对方式的中介作用

目的:探讨应对方式在歧视知觉与男同性恋艾滋病感染者的创伤后应激障碍(PTSD)关系中的中介作用。方法:采用问卷调查法测量145名男同性恋艾滋病感染者的歧视知觉、应对方式和创伤后应激障碍。结果:1歧视知觉与情感中心应对方式、PTSD均呈显著正相关,但与问题中心应对方式无显著相关。2情感中心应对方式在男同性恋艾滋病感染者群体的歧视知觉与创伤后应激障碍之间起部分中介作用。结论:歧视知觉既直接预测男同性恋艾滋病感染者PTSD的症状严重程度,也可以通过情感中心应对方式间接预测其PTSD严重程度。     

KIR and HLA gene combinations in Vogt-Koyanagi-Harada disease

Vogt-Koyanagi-Harada (VKH) disease is a putative autoimmune ocular inflammatory disease and is known to be associated with HLA-DR4 and -DR1 in Mestizos. We examined the genes encoding KIR receptors and human leukocyte antigen (HLA) class I ligands in patients with VKH disease and compared to published controls. We found trends toward more group B KIR haplogroups (p=0.059), with more activating KIR genes, in patients compared to controls. All putative activating KIR-HLA combinations were more common in patients, and some inhibitory KIR-HLA combinations were more common in controls, although the differences were not statistically significant. The trends observed in this study are consistent with those reported for other autoimmune diseases.     

瘢痕疙瘩与HLA-II类基因相关性研究

目的 :探讨免疫遗传学因素在瘢痕疙瘩发病机制中的作用。方法 :用聚合酶链反应 序列特异性引物 (PCR SSP)技术对 5 0例瘢痕疙瘩患者进行HLA Ⅱ类基因分型 ,并以 1 0 0例正常人的HLA Ⅱ类基因为对照。结果 :瘢痕疙瘩组HLA DQ5抗原频率 ( 3 0 % )明显高于对照组 ( 1 4% ) ,相对危险率 (RR) =2 .63 ;HLA DR1 7和HLA DQ8抗原频率 ( 2 % )明显低于对照组 ( 1 4% ,1 5 % ) ,RR =0 .1 3 ,0 .1 2。结论 :瘢痕疙瘩与HLA DQ5基因呈正相关 ,与HLA DR1 7和HLA DQ8基因呈负相关。HLA DQ5、HLA DR1 7和HLA DQ8基因可能是瘢痕疙瘩患者易感的单倍型     

常德市MSM人群艾滋病、性病感染状况调查

目的了解常德市男男性行为者（MSM）人群艾滋病、梅毒和丙型肝炎病毒的感染状况,为制定有效的防治措施提供信息和依据。方法对287例MSM分别进行人类免疫缺陷病毒（HIV）、丙型肝炎病毒（HCV）和梅毒螺旋体（TP）抗体检测。结果 MSM的总感染率为16.4%,其中HIV阳性10份（3.5%）,HCV阳性3份（1.0%）,TP阳性37份（12.9%）。结论常德市MSM人群的HIV和梅毒感染率较高,是艾滋病防治的重点人群,应立即采取积极有效的预防和干预措施。     

HLA in a cohort of Brazilian patients with sarcoidosis

Sarcoidosis is a multisystem granulomatous disorder of unknown etiology. Hereditary etiology has been proposed as a cause of the sarcoidosis, and some Human Leucocyte Antigen (HLA) alleles have been related to the diagnosis or severity of the disease. Löfgren’s syndrome has been linked to patients with the DRB1*03 allele, and non-resolving disease has been associated with the DRB1*07, DRB1*14 and DRB1*15 alleles. However, the results observed in Caucasian patients are not reproducible in other populations, such as in Japanese individuals. The aim of this study is to examine the HLA alleles in Brazilian patients with sarcoidosis confirmed by biopsy.     

Increased level of serum HLA class I antigens in HIV infection correlation with disease progression

Analysis of (sHLA-I) antigens in a large number of HIV-positive subjects found a significant increase of their level, but did not detect any change in their molecular profile. Monitoring at yearly intervals for four years of the sHLA-I antigen level in 14 HIV-positive subjects with a normal sHLA-I antigen level at study entry showed a significant correlation between progressive increase of sHLA-I antigen level and disease progression. Furthermore, a Kaplan-Meier plot of the frequency of development of AIDS in 34 patients whose cases were followed for 7 years showed that sHLA-I antigen level is a strong predictor of progression to AIDS. Its predictive value is comparable to that of serum β₂-μ level, greater than that of serum neopterin, and lower than that of CD4 + T-cell percentage. The predictive value of sHLA-I antigen level in combination with serum β₂-μ level, neopterin level, or CD4 + T-cell percentage is greater than that of each individual variable. These results suggest that measurement of the sHLA-I antigen level may provide useful prognostic information in HIV-positive subjects. Human Immunology 40, 259–266 (1994)     

HIV/AIDS患者CCR5、CXCR4的表达与疾病进展的关系

目的　了解HIV AIDS患者淋巴细胞表面第二受体CCR5、CXCR4的表达 ,分析其与疾病进展的关系 ,探讨HIV感染的免疫基础。方法　收集 33例HIV AIDS患者及 13例健康对照的抗凝全血 ,用流式细胞仪检测第二受体CCR5、CXCR4的表达 ,并分析第二受体表达与病毒载量、CD4 + T淋巴细胞绝对值及T淋巴细胞活化 (HLA DR+ CD38+ )的相关性。结果　艾滋病组CD4 + 、CD8+ T淋巴细胞表面CCR5表达高于无症状HIV 1感染组及健康对照 (P <0 .0 0 1) ;艾滋病组CD8+ T淋巴细胞表面CXCR4表达低于健康对照 (P <0 .0 1)。HIV AIDS患者CD4 + 、CD8+ T淋巴细胞表面CCR5的表达与病毒载量明显正相关 (P <0 .0 1) ;与CD4 + T淋巴细胞绝对值明显负相关 (P <0 .0 1) ,与T淋巴细胞活化(HLA DR+ CD38+ )水平明显正相关 (P <0 .0 0 1)。结论　HIV 1感染者第二受体CCR5的表达与机体对HIV的免疫反应及疾病进展密切相关。     

尿调节素及UMOD基因与慢性肾病的相关性研究进展

尿调节素,又称Tamm-Horsfal蛋白,是由肾小管髓袢升支粗段的小管上皮细胞分泌、也是正常人尿液中含量最多的蛋白。研究发现尿调节素具有多种生物学功能,如调节水盐代谢、调节免疫、防止肾结石产生和防止尿路感染等。近期全基因组关联分析发现编码尿调节素的基因UMOD启动子区单核苷酸多态性与慢性肾病的进展及预后具有很高关联度,并进一步通过孟德尔随机化方法证实尿调节素与慢性肾脏病的因果关系。     

Genotype-phenotype correlation in five Pelizaeus-Merzbacher disease patients with PLP1 gene duplications

Pelizaeus–Merzbacher disease (PMD) is an X-linked myelination disorder most frequently caused by duplication of a genomic segment of variable length containing the PLP1 gene. We studied five PMD male patients affected by the classic PMD form carrying a PLP1 gene duplication. On the basis of clinical and neuroradiological features, two of the five patients appeared to be the most severely affected. In order to establish a possible genotype–phenotype correlation, the extent of the duplication was determined in each patient and in the respective mother by quantifying the copy number of genomic markers surrounding the PLP1 gene by a real-time PCR-based approach. Duplications, ranging in size from 167–195 to 580–700 kb, were in the same genomic interval of the majority of the reported duplications. The extent of the duplicated genomic segments does not correlate with the clinical severity. Interestingly enough, each duplication had one of the two breakpoints in or near to low copy repeats (LCRs), supporting recent evidence concerning a possible role of LCRs in the generation of the duplications in PMD.     

Association of complementary and alternative medicines with HIV clinical disease among a cohort of women living with HIV/AIDS

To assess the association between the use of complementary and alternative medicine (CAM) and HIV clinical disease indicators, CD4+ T-cell counts, viral load, number of HIV-related infections, Centers for Disease Control and Prevention categories, and Karnofsky scores. Data were collected from 391 HIV-positive women aged 18 to 50 years in Alabama and Georgia. A survey examining CAM use and other sociodemographic variables was used. Multiple logistic regression analyses were used to identify predictors of CAM use. Approximately 60% of study participants used 1 or more type of CAM. Predictors of CAM use included higher educational level (odds ratio [OR] = 2.4; P = 0.0008), absence of health insurance (OR = 0.49; P = 0.0055), longer disease duration (OR = 2.21; P = 0.0006), and higher number of infections (OR = 0.58; P = 0.017). Vitamins were the most commonly used CAM ( approximately 36%). Sociodemographic variables associated with vitamin use included higher educational level (OR = 2.34; P = 0.0055), longer disease duration (OR = 1.87; P = 0.026), and higher use among white women than among African-American women (OR = 0.41; P = 0.017). The use of CAM is prevalent among HIV-positive women, and vitamins are the most commonly used CAM among our study population. Several sociodemographic and clinical factors predicted CAM use. These findings have implications for improvement of care for HIV-positive women.     

Frequency and prognostic value of HLA antigens in osteosarcoma patients

A homogeneous group of 53 Caucasian subjects with high-grade osteosarcoma (OS) was typed for HLA-A and B locus antigens. Although no significant differences in the distribution of these antigens were found in comparison with 425 local controls, a trend towards an increase of HLA-B18 and decrease of HLA-B21 was observed. All the patients underwent amputation plus adjuvant chemotherapy and among the 29 patients with a follow-up longer than one year, 9 out of 10 subjects with HLA-A3 antigens developed metastases within a few months. None of the OS patients had the HLA-A3, B7 haplotype which is present in linkage-disequilibrium in the control population.