胸椎黄韧带骨化与HLA-DQA1等位基因的相关性研究

目的分析黄韧带骨化与HLA-DQA1等位基因的相关性.方法用聚合酶链式反应/序列特异引物(PCR-SSP)法对30例OLF患者、51例正常对照进行HLA-DQA1等位基因的基因的分型.结果HLA-DQA1*0401同OLF呈显著正相关(GF=20.0%,RR=7.327,P＜0.01),DQA1*0201与OLF呈显著性负相关(GF=5.0%,RR=0.201,P＜0.01);且DQA1*0401的病因分数＞DQA1*0201的预防分数(EF/PF=0.173/0.162).结论HLA-DQA1*0401可能与OLF的疾病易感性相关,HLA-DQA1*0201可能与OLF的抗性相关,在HLA-DQA1位点存在易感和抵抗双重作用,等位基因之间的共同作用可能是影响OLF发病的重要因素之一.     

慢性乙型肝炎和肝硬化及肝癌与HLA-DQA1基因多态性关系的研究

目的:研究HLA-DQA1等位基因多态性与慢性乙型肝炎(HBV)病毒感染、肝硬化及肝癌的关系。方法:采用聚合酶链序列特异性引物(PCR-SSP)技术分别对168例慢性HBV感染者(包括48例慢性乙型肝炎、42例乙型肝炎肝硬化和78例乙型肝炎后肝癌患者)以及100例对照(感染后自发恢复者)进行HLA-DQA1等位基因的检测。结果:慢性HBV感染者HLA-DQA1*0102的表型频率显著低于对照组(25.6%vs47.0%,OR=0.39,Pc=0.003),DQA1*0601表型频率高于对照组(4.2%vs0,OR=0.96,P=0.039),但后者差异无统计学意义(Pc>0.05)。肝硬化患者DQA1*0104的表型频率显著低于无肝硬化患者(6.4%vs28.4%,OR=0.17,Pc=0.001),DQA1*0201的表型频率高于无肝硬化患者(27.7%vs12.2%,OR=2.76,P=0.014),但后者差异无统计学意义(Pc>0.05)。HLA-DQA1各等位基因的表型频率在肝癌患者与非肝癌患者间差异无统计学意义。结论:HLA-DQA1*0102等位基因可能降低慢性HBV感染的风险,而DQA1*0104等位基因可能降低乙型肝炎肝硬化的风险。乙型肝炎后肝癌的发生与HLA-DQA1等位基因无明显相关性。     

严重多发伤患者单核细胞表面人白细胞DR抗原表达与预后

目的　探讨严重多发伤患者单核细胞表面人白细胞DR抗原表达在创伤病情发展中的作用及与预后的关系。方法　采用直接免疫荧光法流式细胞术对 3 9例严重多发伤患者创伤后的第 1、3、5、7天外周血单核细胞HLA DR抗原表达量进行连续性检测。结果　创伤严重度记分≥2 1分者HLA DR的表达量明显降低 (P <0 .0 1) ,平均荧光道数分别为 (2 2 .2 1± 5 .5 2 )、(18.2 9± 4.2 2 )、(15 .3 7± 3 .76) ;死亡组和多器官功能障碍综合征组HLA DR表达量比康复组明显较少 (P <0 .0 1) ,康复组单核细胞HLA DR水平较治疗前明显升高 (P <0 .0 5或P <0 .0 1)。结论　严重多发伤患者外周血单核细胞HLA DR抗原表达量与创伤严重程度和预后有明显的相关性。     

国人HLA基因与多发性大动脉炎相关性研究

目的：探讨与中国人多发性大动脉炎（TA）相关的白细胞抗原（HLA）基因。方法：采用PCR－SSOP分型技术及群体遗传学方法对44例TA病人及135例健康人的HLA－DQA1及HLA－DQB1位点进行分析。结果：TA组DQA1－0301基因频率（14．8％）较正常人组（30．7％）显著降低（P＜0．05），而TA组DQB10601基因频率（26．1％）较对照组（11．2％）显著升高（P＜0．05），TA病人DQA1－0101－DQB1－0401及DQA1－0103－DQB1－0601单倍型频率升高，而DQA1－0301－DQB1－0401单倍型频率降低，但无显著性差异（P＞0．05），结论：中国人携带HLA Ⅱ－DQB1－0601基因的患TA的可能性显著增加，而携带HLAⅡ－DQA1－0301基因的患TA的机率显著降低。     

黄芪对尿毒症患者血浆sFasL及外周血淋巴细胞亚群状态的作用

目的 :通过测定血浆中sFasL水平和外周血T淋巴细胞亚群状态的分析 ,探讨黄芪对改善尿毒症患者肾功能可能的机制。方法 :采用流式细胞仪和ELISA方法测定 2 5例尿毒症患者黄芪治疗前后及 2 0例正常人血浆中sFasL水平和外周血淋巴细胞亚群状态 ,以及患者治疗前后的肾功能变化。结果 :与正常人比较 ,尿毒症患者血浆中可溶性Fas配体 (sFasL)水平明显升高 ;CD3 CD4 细胞的百分率明显下降 ,CD8 细胞的百分率明显升高 ,CD4 /CD8 的比例小于 1(P <0 .0 1)。 2 5例患者黄芪治疗后血浆中sFasL水平降低 (P <0 .0 5 )。CD4 /CD8 的比例升高 (P <0 .0 1) ;肌酐 (Scr)和尿素氮 (BUN)水平亦有不同程度降低 (P <0 .0 5 )。且动态观察 12例尿毒症患者血肌酐和尿素氮的下降与血浆中sFasL水平的下调呈正相关 (r =0 .5 6 ,P <0 .0 1)。结论 :黄芪能改善尿毒症患者的肾功能 ,提高患者的细胞免疫功能及减慢患者的细胞凋亡是其可能的机制之一。     

IgA肾病患者血与尿血管内皮生长因子水平变化的临床意义

目的 :探讨IgA肾病患者血、尿血管内皮生长因子 (VEGF)水平的临床意义。 方法 :IgA肾病患者2 0例和健康对照 14例 ,留取空腹血清和晨起清洁中段尿 ,采用抗体夹心ELISA方法测定血、尿VEGF水平。详细收集IgA肾病患者的血压、肾功能、2 4h尿蛋白定量和血尿程度等资料 ,与血、尿VEGF对比分析其临床意义。结果 :IgA肾病患者血、尿VEGF水平均高于健康对照组 (P <0 .0 1或P <0 .0 5 ) ;IgA肾病患者血、尿VEGF水平与 2 4h尿蛋白定量呈正相关 (P <0 .0 1或P <0 .0 5 ) ,2 4h尿蛋白定量 <1.5g的患者血、尿VEGF水平明显低于尿蛋白定量 >1.5 g的患者 (P <0 .0 1或P <0 .0 5 ) ;与患者血尿程度、血压无相关性。结论 :IgA肾病患者血、尿VEGF水平与IgA肾病尿蛋白排出量呈正相关     

人类白细胞抗原-DRB1基因多态性与山西省家族性乙型肝炎预后的相关性研究

目的探讨人类白细胞抗原（human lymphoeyte antigen,HLA）-DRB1等位基因多态性与山西省家族性乙型肝炎临床转归及乙型肝炎病毒（HBV）复制状态的相关性。方法采用前瞻性临床流行病学研究方法,以山西省家族性乙型肝炎家庭中的295位成员为研究对象,将其分为健康对照组、慢性无症状携带（ASC）组、慢性乙型肝炎（CHB）组及肝硬化组。采用聚合酶链反应．序列特异性寡核苷酸探针技术（polymerase chain reaction—sequence specific oligonucleotide probe,PCR—SSOP）结合荧光磁珠流式检测技术,进行HLA—DRB1等位基因检测。采用,检验或Fisher’s确切概率计算法比较HLA—DRB1各等位基因频率在各组间以及在HBV DNA不同载量下的分布情况,组间计量资料比较采用方差分析,相关疾病的等位基因风险率以相对危险系数（RR）表示。结果HLA—DRB＊04基因频率在健康对照组为0．159,明显高于ASC组（0．069）和CHB组（0．079）,差异具有统计学意义（χ^2分别为4．892和4．072,P值均〈0．05）;HLA—DRB1＊07等位基因在CHB组和肝硬化组的频率分别为0．131和0．154,明显高于健康对照组（0．049）,差异具有统计学意义（χ^2值分别为4．140和5．529,P值均〈0．05）;HLA—DRB1＊13等位基因频率在健康对照组为0．037,高于CHB组（0）,2组比较差异具有统计学意义（χ^2=3．316,P〈0．05）。HLA—DRB1＊15等位基因频率在ASC组为0．206,在肝硬化组为0．115,2组比较差异具有统计学意义（χ^2=4．287,P〈0．05）。其他各等位基因频率在各组间差异无统计学意义。患者年龄在ASC、CHB及肝硬化3组间的差异有统计学意义（F=33．38,P〈0．01）;HBV DNA阳性组的HLA—DRB1＊07基因频率（0．167）高于HBV DNA阴性组（0．096）（χ^2=5．268,P=0．002）。结论HLA—DRB1＊07与家族性HBV易感性有关,可能是山西省家族性乙型肝炎易感基因或连锁基因。HLA—DRBI＊04及HLA—DRB1＊13与家族性乙型肝炎抗性相关,可能是山西省家族性乙型肝炎抗性基因。     

胰岛素抵抗与尿毒症高血压的关系

目的 :探讨胰岛素抵抗 (IR)与尿毒症患者高血压的关系。方法 :对 4 5例高血压和 14例非高血压尿毒症的患者进行口服葡萄糖耐量试验 (OGTT)和胰岛素释放试验 (IRT) ,计算胰岛素敏感指数 (ISI)、机体糖代谢率 (M )、糖及胰岛素反应曲线下面积 (AUCG、AUCINS) ,分析上述指标与平均动脉压 (MBP)的关系。结果 :(1)尿毒症高血压组糖负荷后的血糖、胰岛素水平、AUCG、AUCINS均显著高于非高血压组 (P <0 .0 5 ) ,而ISI、M低于非高血压组 (P <0 .0 5 ) ;(2 )胰岛素抵抗组 (IR)血压值明显高于非胰岛素抵抗组 (NIR) ,P <0 .0 5 ;(3)尿毒症患者平均动脉压与ISI、M呈负相关 (r =- 0 .5 8、- 0 .5 4 ) ,与AUCG、AUCINS呈正相关 (r =0 .6 2、0 .5 3) ,P≤ 0 .0 0 1。结论 :IR、高胰岛素血症及糖耐量异常 (IGT)与尿毒症患者高血压明显相关 ,可能是引起尿毒症高血压的重要因素之一。     

胸腹部联合损伤患者树突状细胞数量与表面标志的变化

目的　讨胸腹联合损伤后患者外周血树突状细胞 (DC)的变化。方法　胸腹联合损伤患者 ( 2 6例 ,胸腹联合损伤组 ) 2 4h内和健康人 ( 10例 ,对照组 )外周血分离出DC ,通过流式细胞仪检测各组的DC数量 (CMRF -4 4标记法 )及DC表面HLA -DR、CD80 、CD86表达水平以及DC诱导的T细胞反应性增殖。结果　胸腹联合损伤组DC细胞数为 ( 7.5± 3 .3 )× 10 6/L ,明显低于对照组的 ( 14 .9± 5 .1)× 10 6/L(P <0 .0 1)。胸腹联合损伤组DC表面HLA -DR及CD80 、CD86的表达水平与对照组相比明显下降 (P <0 .0 1)。DC诱导的T细胞增殖能力胸腹联合损伤组明显低于对照组 (P <0 .0 1)。结论　胸腹联合损伤患者早期外周血DC数量少 ,功能低下 ,与创伤早期免疫功能低下有关。     

HLA—DRB1基因与IgA肾病关联性的研究

目的：探讨HLA—DRB1等位基因与山西汉族IgA肾病的关联性。方法：用聚合酶链反应序列特异性探针（PCR—SSO）法，对30例IgA肾病患者和45例正常对照者进行了HLA—DRB1等位基因分型。结果：IgA肾病组的HLA—DRB1＊15基因频率明显较正常对照低（5.00％ vs 17.78％，P=0．021，OR=0．243，95％CI：0．068，0.876），IgA肾病组的HLA—DRB1＊04基因频率明显较正常对照高（30．00％ vs 15．56％，P=0．034，OR=2．327，95％CI：1.052，5．145）。结论：HLA—DRB1＊15可能与山西IgA肾病的抵抗性有关，HLA—DRB1＊04可能与山西IgA肾病的易感性有关。     

Association between HLA class II locus and the susceptibility to Artemisia pollen-induced allergic rhinitis in Chinese population.

OBJECTIVE: The aim of the study was to determine whether susceptibility or resistance to Artemisia pollen-induced allergic rhinitis was associated with HLA class II DQA1, DQB1 loci.Study design and setting Forty-one subjects with allergic rhinitis and 41 healthy controls from Beijing were genotyped at HLA class II DQA1, DQB1 alleles by polymerase chain reaction amplification with sequence-specific primers-based technique. RESULTS: The allele frequencies of HLA-DQA1*0201, DQB1*0602 were lower in patients with allergic rhinitis compared with the controls (24.39% versus 46.34%, P = 0.038; 4.88% versus 26.83%, P = 0.007), and the frequency of DQA1*0302 was higher among patients than the controls (58.54% versus 14.63%, P = 0.00004, Pc = 0.0004). CONCLUSION AND SIGNIFICANCE: HLA-DQA1 and -DQB1 genes may be involved in the development of Artemisia pollen-induced allergic rhinitis. HLA-DQA1*0201, DQB1*0602 alleles may be a protective factor and DQA1*0302 may be a susceptible factor for Artemisia pollen-induced allergic rhinitis.     

New DQA1 allele specific antibody against epitope 2D (an exon 1 encoded amino acid). Considerations for alleles under the same P-group designation

The majority of polymorphisms of the Human Leukocyte Antigen (HLA) proteins are clustered at the peptide binding domain (PBD), defined as the area coded by exon 2 and 3 for class I and exon 2 for class II. HLA alleles with the same amino acid (AA) sequence at the PBD are considered functionally equivalent and can be grouped under the same P-group designation. Here we present a case of a kidney recipient, typed as DQA1*01:04, 01:05 and DQB1*05:01P, 05:03, who developed antibodies against all DQ antigens on our Luminex Single Antigen (LSA) panel. Our LSA panel does not include DQA1*01:05 or 01:04, but both alleles belong to the DQA1*01:01P group and beads carrying DQA1*01:01 tested positive. Mature protein sequence alignment demonstrated a single AA mismatch between DQA1*01:04/01:05 and DQA1*01:01 located at position 2 (G vs D), which is encoded by exon 1. Luminex assay by another manufacturer which include a bead carrying patient's own DQA1* type and crossmatch studies with surrogate donors confirmed the presence of an antibody against mismatched epitope 2D. This case illustrates that alleles included in the same P-group may have polymorphisms able to trigger immunological responses and brings attention to the fact that some mature HLA proteins express AA encoded by exon 1, which is structurally part of the PBD.     

A method for HLA-DQA typing by the PCR-SP technique

Abstract There is preliminary evidence that matching for HLA-DQ is important for kidney graft survival. We developed a method for HLA-DQA typing based on the PCR-SP principle. The procedure consisted of three steps: DNA isolation, PCR amplification and visualization of the PCR product under UV light. For the identification of all currently known DQA1 alleles, we designed 18 different primers that allowed typing for the specificities DQA1*0101, *1012, *0103, *0104, *0201, *03, *0401, *0501 and DQA1*0601. For the typing of a single individual, 12 PCR mixes were needed, each containing a primer pair specific for a certain allele group, and a pair of control primers that amplified a non-polymorphic region. The time required for this procedure was approximately 3 h from the time of blood collection. Comparison of this method with DQA typing by the RFLP method in 151 individuals revealed only a single discrepancy. The method can be easily applied for prospective cadaver donor typing.     

Heterozygosity or homozygosity for 2 HLA class II haplotypes predict favorable outcomes for renal cell carcinoma treated with cytokine therapy

PURPOSE: Durable responses to cytokine therapy occur in a small subset of patients with renal cell carcinoma. We determined if a common HLA genotype existed among these patients which might be associated with response and survival. MATERIALS AND METHODS: The study population consisted of 80 patients with metastatic renal cell carcinoma who had received cytokine therapy. DNA obtained from these patients was used for high resolution typing of HLA A, B, C, DRB1, DQA1 and DQB1 alleles. RESULTS: The class II alleles from patients with prolonged disease-free survival were predominantly composed of haplotype DRB1*0301/DQA1*0501/DQB1*0201 and DRB1*1501/DQA1*0102/DQB1*0602. The frequency of heterozygosity or homozygosity for these alleles was significantly greater in the good outcome group of patients than in those whose disease progressed during therapy. Heterozygosity or homozygosity at these loci was also associated with significant prolongation of survival. CONCLUSIONS: We conclude that heterozygosity or homozygosity for the class II haplotypes DRB1*0301/DQA1*0501/DQB1*0201 and DRB1*1501/DQA1*0102/DQB1*0602 is associated with durable response and survival in patients with metastatic renal cell carcinoma treated with cytokine therapy.     

Association between a polymorphism of the transforming growth factor-beta1 gene and genetic susceptibility to ossification of the posterior longitudinal ligament in Japanese patients

STUDY DESIGN: A study was conducted to determine the association between polymorphism of the transforming growth factor-beta1 (TGF-beta1) gene and ossification of the posterior longitudinal ligament (OPLL) prevalence. OBJECTIVE: To examine whether the T869-->C polymorphism of the TGF-beta1 gene is associated with genetic susceptibility to OPLL in Japanese subjects. SUMMARY OF BACKGROUND DATA: In the posterior longitudinal ligament, OPLL is associated with abnormal calcium metabolism. Several candidate genes are associated with the prevalence of OPLL. In the ossified matrix and chondrocytes of adjacent cartilaginous areas of OPLL, TGF-beta1 is overexpressed. METHODS: The TGF-beta1 genotype was identified with an allele-specific polymerase chain reaction method in 319 Japanese subjects (46 subjects with OPLL and 273 control subjects). RESULTS: There was a significant association between the T869-->C genotype and the prevalence of OPLL in the cervical spine. Multivariable logistic regression analysis, adjusted for gender, age, height, and body weight, showed that the frequency of the C allele was significantly higher in subjects with OPLL than in control subjects. CONCLUSIONS: The T869-->C polymorphism of the TGF-beta1 gene is a genetic determinant of a predisposition to OPLL, with the C allele representing a risk factor for genetic susceptibility to OPLL in Japanese subjects. Therefore, TGF-beta1 genotyping may be useful in the prevention of OPLL.     

Particular HLA-DQ alpha beta heterodimer associated with IDDM susceptibility in both DR4-DQw4 Japanese and DR4-DQw8/DRw8-DQw4 whites

Insulin-dependent diabetes mellitus (IDDM) susceptibility is associated with the DR4-DQw4 haplotype in Japanese and the DR4-DQw8/-Drw8-DQw4 genotype (among others) in whites. We investigated whether these Japanese and white individuals encode the same or a similar DQ alpha beta heterodimer, which may be an IDDM-susceptibility molecule in both populations. First, we carried out genomic DQA1 and DQB1 typing with sequence-specific oligonucleotide probes. The results revealed that Japanese DR4-DQw4 and white DR4-DQw8/DRw8-DQw4 IDDM patients carried the DQA1*0301 allele and the DQB1*0401 or DQB1*0402 allele, either in the cis (Japanese DR4-DQw4 individuals) or trans (white DR4-DQw8/DRw8-DQw4 individuals) position. Because the DQB1*0401 and DQB1*0402 alleles differ only at residue 23, these DQB1 genes are very similar. We next tested cells from these individuals with a particular DQ-specific T-lymphocyte clone, HH58. The clone was only restimulated with cells from Japanese individuals who carried the DQA1*0301 and DQB1*0401 alleles in the cis position or white individuals who carried the DQA1*0301 and DQB1*0402 alleles in the trans position. Thus, particular cis- or trans-encoded DQ alpha beta heterodimers, which in both cases are recognized by T lymphocytes, may confer susceptibility to IDDM in both ethnic groups.     

白细胞介素1受体拮抗剂等位基因与紫癜性肾炎和IgA肾病的相关联系

目的 为了进一步阐明紫癜性肾炎(HSPN)及IgA肾病(IgAN)二者在发病机理上可能存在的联系。方法 用PCR方法对43例HSPN,97例IgAN患者和98名正常人IL-l受体拮抗剂(IL-lra)基因数目可变的串联重复(VNTR)多态性进行了分析。结果 HSPN患者白细胞介素l受体拮抗剂等位基因(IL IRN 2)的携带率明显高于正常人(P<0.02)和IgAN患者(P<0.05)。在IgAN患者中表现为反复发作性肉眼血尿者其IL IRN 2等位基因的携带率明显高于其它临床类型IgAN患者(P<0.o1),而与HSPN无显著性差异(P>0.05)。结论 HSPN患者IL-lra基因特定的遗传背景在其发病机理中可能起一定作用。IgAN患者中表现为反复发作性肉眼血尿者较其它临床类型IgAN患者与HSPN有更多的相似之处,而ILlRN 2等位基因高携带率可能是二者发病机理的共同点之一。     

HLA DRβ1 Alleles in Pakistani Patients with Rheumatoid Arthritis

Objective: To determine frequencies of HLA DRβ1 alleles in rheumatoid arthritis in Pakistani patients. Study Design: Cross sectional / analytical study. Place and Duration of Study: Department of Immunology, Armed Forces Institute of Pathology, Rawalpindi in collaboration with Rheumatology departments of Military Hospital, Rawalpindi and Fauji Foundation Hospital, Rawalpindi, from January 2009 to January 2010. Methodology: HLA DRβ1 genotyping of one hundred Pakistani patients, diagnosed as having RA as per American College of Rheumatology revised criteria 1987, was done. HLA DRβ1 genotyping was carried out at allele group level (DRβ1*01-DRβ1*16) by sequence specific primers in RA patients. Comparison of HLA DRβ1 allele frequencies between patients and control groups was made using Pearson's chi-square test to find possible association of HLA DRβ1 alleles with RA in Pakistani rheumatoid patients. Results: HLA DRβ1*04 was expressed with significantly increased frequency in patients with rheumatoid arthritis (p <0.05). HLA DRβ1*11 was expressed statistically significantly more in control group as compared to rheumatoid patients indicating a possible protective effect. There was no statistically significant difference observed in frequencies of HLA DRβ1 allele *01, DRβ1 allele *03, DRβ1 allele *07, DRβ1 allele *08, DRβ1allele *09, DRβ1 allele*10, DRβ1 allele *12, DRβ1 allele *13, DRβ1 allele *14, DRβ1 allele *15 and DRβ1 allele *16 between patients and control groups. Conclusion: The identification of susceptible HLA DRβ1 alleles in Pakistani RA patients may help physicians to make early decisions regarding initiation of early intensive therapy with disease modifying anti rheumatic medicines and biological agents decreasing disability in RA patients.