全身性红斑狼疮、银屑病和胰岛素依赖性糖尿病病人补体成份BfSS亚型的检测

用薄层聚丙烯酰胺凝胶等电聚焦方法对62例全身性红斑狼疮(SLE)、61例寻常型银屑病(PV)及17例胰岛素依赖性糖尿病(IDDM)病人补体第二途径成份B因子SS型的亚型分布及基因频率进行了检测。结果表明,SLB病人BfSS亚基因频率分别为,S_A0.516,S_B0.484;PV为:S_A0.492,S_B0.508,IDDM为:S_A0.676,S_B0.324。与正常人(S_A0.414,S_B0.586)比较,三种疾病的S_A亚基因频率均有不同程度的增高,并伴有S_B亚基因频率的降低。统计结果表明,IDDM与正常人的差异显著(P<0.01),SLE次之(P<0.05),PV不著(P>0.1)。此外,这三种疾病病人BfS_A-S_A亚型的表型频率(分别为30.6%、31.1%和41.1%)都显著高于正常19.3%),统计学表明差异均显著(P<0.05)。     

胰岛素依赖型糖尿病患者ApoE表型与基因型差异

为了研究胰岛素依赖型糖尿病（IDDM）患者载脂蛋白（Apo）E表型与基因型的差异，选择了中国汉族IDDM患者54例。采用等电聚焦法(IEF)和PCR/ASO探针杂交及扩增不应突变系统(ARMS)分别检测其ApoE表型和基因型。结果显示ApoE表型与PCR/ASO法分析的ApoE基因型差异14例(25.9%)，与ARMS法分析的ApoE基因型差异12例(22.2%)（P＜0.01）。相比之下，ApoE2异型频率相对增高(16.67%)。从而推断IDDM患者ApoE转录后的化学修饰可导致ApoE表型与基因型不一致。     

HLA和胰岛素依赖型糖尿病

本文对35名胰岛素依赖型糖尿病病人(IDDM)及53名正常人进行了HLA分型。分型试剂由第二届亚洲大洋洲组织相容性专题讨论会提供包括HLA-A、B、C和DR。同时测定了第六条染色体的遗传标志包括GLO、Bf和C2的变异体。 结果发现病人组A11和CW4的表现型频率减低,校正后仍有显著意义。病人组DR3和DRW9表现型频率与对照组比较有增高,特别是DRW9校正后仍然有非常显著的意义(校正p值为0.00062)。 我们的结果提示中国人IDDM病人可能存在二个HLA-DR抗原与IDDM相关联。     

抗胰岛细胞抗体:Ⅰ型糖尿病的免疫学标志

IDDM是一种自身免疫器官特异性疾病.在前驱期和发病后短期内病人血清中可检测出抗胰岛细胞胞浆抗体(ICA),ICA检测已用于IDDM一级亲属及普通学龄儿童的IDDM预测.多数IDDM发病时血清ICA阳性.部分Ⅱ型糖尿病(NIDDM)病人ICA也呈持续阳性,预示为缓慢发病的IDDM.ICA还可用于IDDM病人免疫抑制治疗及胰腺或胰岛移植治疗的疗效判断.     

C反应蛋白酶联免疫分析和化学发光免疫分析的建立及其初步应用

采用高纯度的C反应蛋白(CRP)免疫兔,获取高效价的CRP抗体.提取抗血清中IgG,包被酶标板.用生物素标记CRP抗原,辣根过氧化物酶标记亲和素,建立CRP的ELISA和化学发光免疫分析法(CLIA).用ELISA测定正常人80名,心梗患者43例和重症监护室(ICU)病人39例的血清CRP水平,发现心梗患者(27.21±38.67μg/mL)明显高于正常人(0.57±2.99μg/mL)(P《0.05);ICU病人(117.48±97.05μg/mL)明显高于正常人及心梗患者(P《0.001和P《0.01).用CLIA检测正常人68名,结果为0.77±3.29μg/mL.随机收集病人血清样品73份,分别用免疫比浊法、ELISA和CLIA检测,经统计三种方法高度相关.结果提示:ELISA和CLIA是一种简便、灵敏和特异的CRP测定方法,可用于人血清中CRP的测定.     

HBsAg无症状携带者T、B细胞数及血清花环形成抑制因子的观察

本文用抗胸腺细胞血清细胞毒试验检测了29例HBsAg无症状携带者外周血T细胞数,与正常人(20例)相比显著低下(P<0.001);而用Et法检测的结果与正常人(20例)相比也显著低下(P<0.001)。用FBC花环试验检测19例HBsAg无症状携带者外周血B细胞数,与正常人(20例)相比无显著差别(P>0.06)。此外,HBsAg无症状携带者血清中花环形成抑制因子(RIF)阳性为6/20例(30％)。     

免疫治疗与放、化疗联合疗法的淋巴细胞表型及临床疗效研究

目的探讨恶性肿瘤患者进行放、化疗联合免疫治疗对淋巴细胞表型的影响及其与临床表现的关系。方法共42例晚期肿瘤病人，其中22例在放、化疗的同时输入共刺激的正常人外周血淋巴细胞(PBLs)进行免疫治疗，每周2次，8次为一疗程。另20例病人作为对照组，仅接受放、化疗。治疗前后用流式细胞仪检测淋巴细胞表型并观察临床症状。结果放、化疗联合免疫治疗组22例病人的淋巴细胞表型与治疗前比较CD+3、CD+4、CD+4/CD+8有升高，CD+8降低，但无显著差异，PS评分改善，P＜0.05；与20例对照组比较，淋巴细胞表型治疗前后均无显著差异，但治疗后PS评分改善，P＜0.05。放、化疗联合免疫治疗组病人中有15例(68.18%)的淋巴细胞表型明显改善，CD+3、CD+4治疗后明显升高，与治疗前比较P＜0.05，CD+95升高，P＜0.02，PS评分改善，P＜0.01；与20例对照组相比，治疗前各项无显著差异，治疗后CD+4升高，P＜0.05，PS评分改善，P＜0.05；放化疗联合免疫治疗病人中有7例(31.82%)的淋巴细胞表型改善不明显，与治疗前及对照组比较，均无显著差异，但PS评分改善，P＜0.02。对照组淋巴细胞表型与临床症状改变均不明显。结论免疫治疗与放化疗联合治疗，可能有利于减轻放化疗的副作用，提高疗效，改善生活质量。     

癌症患者T细胞集落形成及血清免疫抑制作用

采用T细胞集落甲基纤维素微量培养方法,发现20例癌症病人外周血T细胞集落(CFU-TL)形成能力显著低于正常人(P<0.001)。20例病人的血清对自身及正常人的CFU-TL均有明显抑制作用(P<0.001,P<0.001),抑制率分别为41.33±25.30%和47.96±22.94%。16例经手术切除肿瘤病人术后7至10天中,CFU-TL产率较手术前有所提高(P<0.01),同时,病人的血清对自身及正常人CFU-TL抑制作用明显减弱(P<0.0025,P<0.0025),术后病人CFU-TL产率提高的程度与病人血清抑制率下降的程度呈正相关关系(r=0.8024,P<0.0005)。结果表明:癌症病人的T细胞集落形成能力明显降低,部分原因由病人血清中存在免疫抑制物质所致。     

炎性关节炎患者调节性T细胞亚群的变化及其对TNF-α拮抗剂治疗的反应

人外周血CD4+Foxp3+调节性T细胞(regulatory T cells,Treg)的表型和功能存在异质性。本文用Foxp3和CD45RO共同标记Treg,研究强直性脊柱炎(AS)和类风湿性关节炎(RA)患者外周血Treg亚群数量以及接受TNF-α拮抗剂治疗对其亚群的改变。分别采集未经治疗的21例活动性AS患者和27例活动性RA患者的外周血。两种疾病各有14名患者接受TNF-α拮抗剂治疗40周。应用流式细胞术检测外周血Treg细胞数量。在CD4+T细胞中,活动性AS和RA患者其表型为CD45RO+Foxp3hi的效应型Treg(AS与正常对照相比,P=0.020;RA与正常对照相比,P0.001)和表型为CD45RO-Foxp3low的幼稚型Treg(AS与正常对照相比,P=0.009;RA与正常对照相比,P=0.001)细胞数量均显著下降。与正常对照相比,AS和RA患者的CD4+Foxp3+T细胞中非Treg亚群细胞(表型为CD45RO+Foxp3low)的比例显著增加(AS与正常对照相比,P=0.027;RA与正常对照相比,P0.001)。TNF-α拮抗剂治疗后,AS患者外周血中效应型Treg(P=0.025)和幼稚型Treg(P=0.005)细胞数量均增加,RA患者中仅效应型Treg细胞数量增加(P=0.003)。因此,不仅分析CD4+Foxp3+T细胞总数,而且分析其中不同表型的亚群细胞数量更有助于明确炎性关节炎的发病机制。     

葡萄糖-6-磷酸酶是一种潜在类风湿关节炎新自身抗原

目的检测类风湿关节炎(rheumatoid arthritis,RA)早期和晚期患者血清中葡萄糖-6-磷酸酶水平,探讨葡萄糖-6-磷酸酶与RA的相关性及其诊断意义。方法患者血清和正常人血清采用二维凝胶电泳免疫印迹的比较寻找差异蛋白点,质谱测序和Western bolt证明患者血清葡萄糖-6-磷酸酶在患者高表达在正常人则检测不到。在此基础上,分别对早期、晚期、非RA患者(其它自身免疫性疾病)和正常人20例、40例、30例、40例血清通过ELISA的方法检测葡萄糖-6-磷酸酶水平,分析血清葡萄糖-6-磷酸酶水平与RA的相关性。结果 RA血清中的葡萄糖-6-磷酸酶水平明显高于正常人(P0.001)和其它自身免疫病患者(P0.001);进一步比较与RA现有诊断指标的相关性,发现葡萄糖-6-磷酸酶血清水平与ESR、CCP、DAS28评分具有显著相关性(P0.05)而与RF无显著相关(P0.05);与临床皮质激素治疗治疗呈负相关。结论提示葡萄糖-6-磷酸酶是RA相关抗原,其血清水平对RA可能具有早期诊断价值;与激素治疗负相关说明该指标反映了疾病活动度;其在RA疾病发生及进展中的作用机理有待于进一步研究。     

全身性红斑狼疮病人补体第二途径B因子多态性的检测

用琼脂糖凝胶高压电泳继以单相抗B因子血清免疫固定技术对81例份全身性红斑狼疮(SLE)病人血清进行了补体第二途径B因子(Bf)遗传多态性的检测。检测结果表明,SLE病人的BF F型的基因频率(0.1914)显著高于正常人(0.1159),P_(修正)<0.05。F型阳性人群SLE发病的相对危险性(RR)为1.79,亦具统计学意义(P<0.05)。     

Properdin factor B and complement factor C4 allotypes in rheumatoid arthritis: results of a follow-up study.

The association between allotypes of properdin factor B (Bf), the fourth component of complement (C4A and C4B), and rheumatoid arthritis (RA), was investigated in a well-characterized cohort of RA patients who were followed from an early phase of the disease for a mean duration of 6 years. The frequencies of probable heterozygous C4AQ0 and of C4A3 were lower in RA patients compared to controls, irrespective of the presence of DR4 [relative risk (RR): 0.52 and 0.49, respectively, 95% confidence intervals (95% CI): 0.34-0.80 and 0.29-0.82]. The frequency of C4A4 was higher in RA patients compared to controls (RR: 1.86, 95% CI: 1.03-3.35), especially in DR4 positive RA patients compared to DR4 positive controls (RR: 2.58, 95% CI: 1.07-6.25), indicating a positive association of this allotype with RA additional to DR4. Bf and C4B allotypes were comparable in RA patients and controls. We did not find significant differences in Bf and C4 allotype frequencies in RA patients subdivided according to severity of the disease into a mild group and a progressive group. Because of inconsistent results in all studies on Bf and C4 allotypes, we conclude that C4 and Bf allotypes do not seem to have an important independent effect on determining disease susceptibility.     

HLA, A, B, C, DR, C4, Bf in insulin-dependent diabetics in the Tunisian population

There is no doubt that the autoimmune process in human disease depends on genetic factors. Varying associations were noticed between HLA DR and autoimmune disorders. The frequency of HLA-A-B and DR antigens as well as the Bf and C4 allotypes have been investigated in insulinodependant diabetes mellitus (IDDM) and compared to that of healthy controls in Tunisian population. An increase of A30, DR3, DR4, BfF1, C4AQ0 and C4BQ0 and decrease of B40, DR2, DR5 and DR6 were found in diabetes when compared to the value observation controls. The strongest association was noticed with HLA, DR3 and DR4. The prospective role of DR2 and DR5 antigens were also confirmed. Examination of HLA, Bf and C4 alleles. Two supratypes associated with IDDM have been observed among the Tunisian patients.     

HLA antigens and complotypes in insulin-dependent diabetes mellitus

One hundred and thirty-six Finnish patients with insulin-dependent (type I) diabetes mellitus were investigated for the HLA-A, B, D and DR antigens as well as the Bf and C4 allotypes. The statistically significant increase in the frequencies of HLA-A9, B8, B15, Dw3, Dw4, DR3, DR4, C4A0 and C4B3 was observed when compared with the healthy controls. About 79% of the patients had HLA-DR4, and 53% had HLA-DR3 antigens. A rare C4 allele C4B3 was found in 21% of the patients, whereas only in 2% among the controls (relative risk 16.35). The etiological fraction (EF) values indicated that HLA D/DR alleles were the best markers for IDDM, the observed EF for HLA-DR4 in diabetes was as high as 0.70. Examination of HLA, Bf and C4 phenotypes suggested that at least two supratypes "B15 BfS C4A3B3 D(R)4" and "B8 BfS C4A0B1 D(R)3" were markers for the susceptibility to type I diabetes, one third of our patients had either of these supratypes. The protective role of DR2 and Dw2 antigens was also confirmed: no HLA-Dw2 positive patients and only one with HLA-DR2 was found.     

HLA-A, B, C, DR antigens, Bf, C4 and glyoxalase I (GLO) polymorphisms in French Basques with insulin-dependent diabetes mellitus (IDDM)

The Basques were previously shown to present a high frequency of HLA—B18 and Bf Fl. which are known to be associated with insulin dependent diabetes mellitus (IDDM). During the VIII International Histocompatibility Workshop, we studied HLA-A, B, C, DR; Bf, C4 and GLO.I polymorphisms in 51 unrelated French Basque IDDM patients and in 50 controls. Haplotypes were established by family studies in all controls and some patients. Two haplotypes were frequently found in the controls: HLA- A1, Bw57, Bf S, C4 FIS, DR7 and HLA- Aw30, Cw5, B18, Bf Fl, C4 Fs°, DR3. The first one was not found in the patients. All the components of the second haplotype had increased frequencies possibly as a consequence of linkage disequilibrium with HLA— DR3 : a highly significant association between IDDM and HLA-DR3 was observed (90.2% vs 24.0%, relative risk (RR) = 29.1. P < 10⁻¹¹). The HLA-DR4 frequency was slightly increased (37.3% vs 16.0%). and HLA—DR2 was not found. The silent allele C4 s ° was particularly associated with early diagnosed IDDM (86.7% in patients with age at onset under 20 years vs 57.1% in other patients, P < 0.02). The high relative risk for HLA-DR3/DR4 heterozygous vs that of individuals, possibly HLA-DR3 homozygous, supported the hypothesis that two HLA-DR linked genetic factors could be involved in the inheritance of IDDM susceptibility.     

Graves氏病人血清补体第二途径B因子遗传多态性的检测

1972年Alper等人用琼脂糖凝胶电泳加免疫固定首先发现人群中补体第二途径B因子(Properdin Factor B,Bf)呈现遗传多态性,他们当时描述了四种Bf表型,常见的S(从slow)与F(fast),及少见的SO7(slow than S)与Fl(fast than F),并提出Bf表型受常染色体共显性等位基因控制。十余年来,各国学者对Bf的多态现象进行了     

Complement C2, C3, C4 and factor B allele distribution in the Gipsy population in Hungary

Allotype frequencies of four complement proteins (C3, C2, factor B, and C4) were tested in 150 healthy Hungarian and 126 healthy Gipsy individuals living in Hungary. We observed significant differences between the two ethnic groups in the incidence of C3*F, Bf*F, C4A*Q0, C4A*3, C4B*1 and C4B*2 allotypes. Bf*F occurred more frequently among Gipsies, while frequencies for the other three allotypes was lower in this group than in Hungarians. The similarities in the allotype frequencies of C3 and Bf among Gipsy and Gaddis (India) populations supports the Indian origin of the former ethnic group.     

Association of HLA-Bw65 with two major complotypes

The association between the HLA-B14 subtypes Bw64 and Bw65 and complement allotypes (C2, Bf and C4) was investigated in both population and family studies. Bf, C4A and C4B allotyping was performed on 37 Bw64 and 35 Bw65 positive unrelated Welsh/English subjects. Sixteen HLA-Bw65 bearing haplotypes were characterized for HLA-ABC, DR and DQ antigens and complement allotypes, including C2. The findings of the population study suggested that the complement haplotype associated with Bw64 is BfS, C4A2, C4B2. The population and family studies revealed two major complement haplotypes associated with HLA-Bw65: (i) C2C, BfF, C4A3, C4A1 - often associated with HLA-A3, Cw8 and DRw13, and (ii) C2C, BfS, C4A2, C4B2 - often associated with HLA-Aw33, Cw8 and DR1 or with A28, Cw8 and DRw13. The HLA-Bw65 bearing haplotypes of three families carried a C4B2B1 duplication of the C4B locus. In these families three C4B gene products were identified in the Bw65 positive members using an anti-C4B monoclonal antibody. It is suggested that most, if not all, HLA-Bw65 bearing haplotypes may possess a C4B locus duplication.     

贵州苗族补体Bf和C4B同种异型的多态性

用高压琼脂糖电泳后的免疫固定试验，检测了贵州苗族健康成人的补体旁路Ｂ因子（Ｂｆ）和Ｃ４Ｂ的同种异型。在检出的基因型中Ｂｆ有３个，Ｃ４Ｂ有１０个，最高基因频率的基因是Ｂｆ＊Ｓ０．９４４４。Ｃ４Ｂ＊２０．５０００．     

HLA－B51与中国北方汉族中的白塞氏病相关联

对１２０例白塞氏病患者和１００名无关健康人进行了ＨＬＡ－Ａ、－Ｂ、－Ｃ、－ＤＲ和－ＤＱ抗原分型检测，并使用Ｌｙｍｐｈｏ－Ｂ－Ｋｗｉｋ分离出Ｂ－淋巴细胞进行ＨＬＡ－ＤＱ和－ＤＲ分型，用琼脂糖电泳，免疫固定等技术测定了Ｂｆ和Ｃ４同种异型。其中，５５．８３％（６７／１２０）患者中检出ＨＬＡ－Ｂ５１，而对照组仅１２％。Ｘ２和ＲＲ分别为４５．５４和９．２７（Ｐ＜０．０００５）．完全型组ＨＬＡ－Ｂ５１更为常见（６２．７９％）；除患者组的Ｃ４ＡＱ０明显高于对照组外，其它的ＨＬＡ抗原、Ｂｆ和Ｃ４同种异型无明显的组间差异。结果提示：某些内在因素，诸如免疫遗传背景（存在有与ＨＬＡ－Ｂ５１密切相关的ＢＤ易感基因）在其病因及发病机理中可能起着重要作用。