HLA-B2704转基因小鼠模型的建立

目的　制备HLA B2 70 4转基因小鼠。方法　应用显微注射将HLA B2 70 4基因注入C5 7BL/6×昆明鼠和昆明鼠×昆明鼠F1代受精卵 ,出生动物及其后代经PCR初筛 ,采用斑点杂交和Southern杂交对阳性标本作进一步鉴定和整合基因拷贝数测定 ,阳性者行免疫组化染色和流式细胞术检测HLA B2 70 4蛋白表达水平。结果　 10只仔鼠 (首建转基因鼠 ,founder)表达有HLA 2 70 4基因。由注射基因后移卵出生的 10 1只小鼠中 ,C5 7BL/6×昆明鼠杂种 3 7只 ,阳性 7只 ( 18 9% ) ,昆明鼠×昆明鼠杂种 64只 ,阳性 3只 ( 4 7% ) ,阳性动物外周血淋巴细胞、皮肤和胃肠道上皮细胞内均有HLA B2 70 4蛋白表达。结论　应用C5 7BL/6和昆明鼠成功建立了HLA B2 70 4的转基因小鼠。与昆明鼠×昆明鼠F1代相比 ,C5 7BL/6×昆明鼠F1代更适合用于转基因动物制备。     

HLA—B2704转基因小鼠自发性前葡萄膜炎

目的 观察ＨＬＡ－Ｂ２７０４转基因小鼠葡萄膜炎发病情况和糖皮质治疗的初步疗效。方法 应用ＰＣＲ、斑点杂交、Ｓｏｕｔｈｅｒｎ杂交、免疫组织化学和流式细胞术进行阳性动物筛选和表达检测。隔日观察眼部症状，可疑动物行眼底镜以及裂隙灯检查，眼球组织切片行ＨＥ染色，房水行细胞计数和分数统计。结果 部分ＨＬＡ－Ｂ２７０４转基因阳性鼠出现明显畏光，眼科检查发现角膜后沉淀。病理发现角膜上皮细胞增生，排列紊乱。房水     

HLA-B2704转基因小鼠自发性前葡萄膜炎

目的　观察HLA B2 70 4转基因小鼠葡萄膜炎发病情况和糖皮质激素治疗的初步疗效。方法　应用PCR、斑点杂交、Southern杂交、免疫组织化学和流式细胞术进行阳性动物筛选和表达检测。隔日观察眼部症状 ,可疑动物行眼底镜以及裂隙灯检查 ,眼球组织切片行HE染色 ,房水行细胞计数和分类统计。结果　部分HLA B2 70 4转基因阳性鼠出现明显畏光 ,眼科检查发现角膜后沉淀。病理发现角膜上皮细胞增生 ,排列紊乱。房水总细胞数增加 ,炎性细胞与总细胞数之比为9 2 4%。正常对照比值为 0 75 %。 χ2 检验显示二者差异存在显著性 ,P <0 0 5。地塞米松治疗可以减轻症状 ,停药后症状加重。符合前葡萄膜炎诊断。结论　首次在转基因小鼠观察到类似人的HLA B2 7相关性前葡萄膜炎。     

银屑病关节炎临床特点及与HLA—B27的相关性研究

目的 分析银屑病关节炎（ＰＡ）的临床特点及与ＨＬＡ－Ｂ２７的相关性。方法 选５５例住院患者，总结分析其皮疹和关节炎，关节外表现，Ｘ线特征及骶髂关节炎与ＨＬＡ－Ｂ２７的关系。结果 ３６例皮疹先于关节炎出现，１３例关节炎在先，６例同时发生。３８例有指甲改变。手足小关节最易受累，共计４５例，占８１．８％，２８例（６２．２％）伴远端指间关节（ＤＩＰ）受累。２８例有关节外表现。２０例有手足小关节Ｘ线破坏征，     

以关节炎为首发表现的银屑病关节炎的临床特征

目的　分析以关节炎为首发表现的银屑病关节炎 (PA)的临床特征。方法　对 15例以关节炎为首发表现的PA的临床体征、实验室及放射学特点进行分析。结果　 15例PA患者中 ,男性 11例 ,女性 4例 ,男女之比为 2 75∶1。发病年龄 11～ 4 7岁 ,平均 (2 4± 13)岁 ,病程 2个月～ 30年 ,平均 (11± 15 )年 ,关节炎和皮疹间隔时间为 2个月～ 30年 ,平均 (9± 9)年。HLA B2 7阳性者 13例 ,有骶髂关节炎者 10例。结论　以关节炎为首发表现的PA患者骶髂关节受累多 ,并与HLA B2 7有一定相关。     

HLA-B27转基因动物研究进展

ＨＬＡ Ｂ2 7是第一个发现与疾病相关的ＨＬＡ等位基因 ,也是到目前为止相关性最高的一个。自 1973年Ｂｒｅｗｅｒｔｏｎ等[1] 和Ｓｃｈｌｏｓｓｔｅｉｎ等[2 ] 提出强直性脊柱炎 (ａｎｋｙｌｏｓｉｎｇｓｐｏｎｄｙｌｉｔｉｓ ,ＡＳ)与ＨＬＡ Ｂ2 7抗原有非常强的关联以来 ,人们发现ＨＬＡ Ｂ2 7还与反应性关节炎、银屑病性关节炎、炎症性肠病性关节炎、青少年脊柱关节病、急性前葡萄膜炎等明显相关。随着研究的深入 ,虽然人们提出了许多ＨＬＡ Ｂ2 7与ＡＳ相关的假说 ,如分子模拟假说、关节源性肽假说、Ｂ2 7自身的变异等 ,但是Ｂ2 7在Ａ…     

HLA-B27相关性反应性关节炎的发病机制

ＨＬＡＢ２７相关性反应性关节炎的发病机制已有二条线索：首先，关节炎由某些细胞内寄生菌感染诱发，包括鼠伤寒沙门菌、福氏志贺菌、小肠结肠耶尔森菌和沙眼衣原体；其次，多数患者ＨＬＡＢ２７阳性。由于ＨＬＡＢ２７在其相关性疾病—强直性脊柱炎的阳性率很高，...     

HLA-B27阴性与阳性强直性脊柱炎的对比研究

了解ＨＬＡ－Ｂ２７在强直性脊柱炎发病及病情活动中的作用。方法；对比分析５１例ＨＬＡ－Ｂ２７阴性，ＡＳ患者和１０２例病程相同的ＨＬＡ－Ｂ２７阳性ＡＳ患者的临床表现及实验室检查结果。结果；Ｂ２７组与Ｂ２７组最明显的差异是前者以女性多见，平均发病年龄偏晚，且临床上较少出现全身症状，外周关节炎及严重的髋关节病变，但合并炎风湿关节炎明显多于阳性组，家族聚集现象，虹膜睫状体炎和脊柱的Ｘ线片改变在两组无差异。     

银屑病关节炎临床特点及与HLA-B27的相关性研究

目的分析银屑病关节炎（ＰＡ）的临床特点及与ＨＬＡＢ２７的相关性。方法选５５例住院患者,总结分析其皮疹和关节炎、关节外表现、Ｘ线特征及骶髂关节炎与ＨＬＡＢ２７的关系。结果３６例皮疹先于关节炎出现,１３例关节炎在先,６例同时发生。３８例有指甲改变。手足小关节最易受累,共计４５例,占８１８％,２８例（６２２％）伴远端指间关节（ＤＩＰ）受累。２８例有关节外表现。２０例有手足小关节Ｘ线破坏征,１９例出现骶髂关节炎或脊柱炎。３８例检测ＨＬＡＢ２７,１９例（５０％）为阳性,其中有骶髂关节炎者为８４２％（１６／１９）。结论伴ＤＩＰ受累的寡关节炎是ＰＡ的特征性表现,中轴关节受累常无症状,并与ＨＬＡＢ２７密切相关。治疗应选择对皮疹及关节炎均有效的甲氨蝶呤或环孢霉素Ａ,以及包括金制剂、柳氮磺胺吡啶等在内的联合方案。     

与HLA-B27相关的关节炎的动物模型

与ＨＬＡＢ２７相关的关节炎的动物模型黄烽李胜光余得恩与ＨＬＡＢ２７相关的关节炎是一组关节的炎性疾病，主要包括强直性脊柱炎和赖特综合征。虽然迄今在强直性脊柱炎尚未分离出已知病原，但在部分赖特综合征病人，关节炎发病前常有肠道和泌尿系感染，其病原菌包括...     

Frequency of HLA-B27 alleles in Brazilian patients with psoriatic arthritis

This prospective study analyzed the frequency of HLA-B27 and its alleles in 102 Brazilian patients with psoriatic arthritis (PsA). The association of the HLA-B27 alleles with these variants was compared to a control healthy HLA-B27 positive group of 111 individuals. There was a predominance of male gender (59.8%), Caucasian race (89.2%), and negative HLA-B27 (79.4%) patients. Asymmetric oligoarthritis (62.7%) was the most frequently observed clinical PsA subgroup, followed by spondylitis (16.7%), and polyarthritis (15.7%). Male gender and the spondylitis subgroup were statistically associated to the positive HLA-B27, and the oligoarthritis subgroup was associated to the negative HLA-B27. Among the 21 HLA-B27-positive PsA patients, there was a significant prevalence of the HLA-B*2705 allele (90.5%), similar to that observed in the control group (80.2%); HLA-B*2703 and HLA-B*2707 were statistically associated to the control group.     

人类白细胞抗原B2704基因在转基因小鼠中的整合与表达

目的研究人类白细胞抗原(HLA)-B2704基因在转基因小鼠中的整合与表达。方法应用显微注射技术将HLA-B2704基因注入C57BL/6×昆明鼠和昆明鼠×昆明鼠F0代受精卵,出生动物进行聚合酶链反应(PCR)筛选及Southern杂交鉴定。阳性者进行反转录(RT)-PCR和流式细胞光度术检测其在mRNA和蛋白水平的表达。结果101只F0代仔鼠中,10只F0代仔鼠基因组中有HLA-B2704基因整合。其中昆明鼠×昆明鼠F0代3只,整合率为4.7%,C57BL/6×昆明鼠F0代7只,整合率为18.9%(字2检验,P<0.05)。阳性小鼠的皮肤、结肠、睾丸和脾脏组织中均有HLA-B2704基因的mRNA水平表达,其外周血淋巴细胞膜上HLA-B2704基因的蛋白表达为7.87%(直方图统计表,5%以上为阳性)。结论尽管受精卵的注射成活率差异无统计学意义,在注射浓度相同的情况下,含有C57BL/6基因背景的受精卵更易整合外源基因,而单纯昆明鼠的受精卵不易接受外源基因。C57BL/6×昆明鼠F0代比单纯昆明鼠F0代更适合于转基因动物的制备。HLA-B2704的重链分子与小鼠茁2微球蛋白结合的复合物不稳定,不易于在细胞表面表达。     

Risk of haematological malignancies in HLA-B27 carriers

Human leucocyte antigen (HLA)-B27 carriers are predisposed to inflammatory and autoimmune diseases. Among 1137 patients with haematological diseases, 59 HLA-B27 carriers were identified. Compared with 18 774 volunteers, the incidence of HLA-B27 was increased in patients with acute leukaemia (relative risk RR = 1.67, P = 0.002), for both acute myeloid leukaemia (AML) (RR) = 1.67, P = 0.007) and acute lymphoblastic leukaemia (ALL) (RR = 1.68, P = 0.094). Of all the HLA-B27 carriers, four patients had ankylosing spondylitis (AS), all with lymphoid malignancies (three ALL, one Hodgkin's disease), whereas no HLA-B27 carriers with myeloid leukaemia had AS symptoms (P = 0.006). This suggests that HLA-B27 carriers may have an increased risk of acute leukaemia and those with concomitant AS may be predisposed to lymphoid malignancies.     

Beta 2-microglobulin amyloid deposit in HLA-B27 transgenic rats

Abstract

Beta 2-microglobulin (beta 2-m) amyloid deposition develops serious orthopedic complications in patients with long-term hemodialysis. However, the exact pathogenesis of amyloid deposition in patients with dialysis is unknown. We used transgenic rats with HLA-B27 and beta 2-m to develop an in vivo animal model with beta 2-m amyloid-associated disorders to investigate the mechanism of formation and to report histological findings of beta 2-m amyloid deposition. Transgenic rats were divided into two groups: group 1, no treatment; group 2, arthritis induced by type II collagen. A large number of cells labeled with beta 2-m were observed in the bone marrow and synovium of the knee joint in transgenic rats. In addition, amyloid deposition, identified by Congo red staining, was found only in the knee joints of the transgenic rats with collagen-induced arthritis. Immunostaining with beta 2-m demonstrated the same pattern of tissue distribution as Congo red in serial sections. We hypothesized that elevated serum beta 2-m level-associated local chronic inflammation leads to the development of amyloid deposits and resultant arthropathy.     

Beta 2-microglobulin amyloid deposit in HLA-B27 transgenic rats

Beta 2-microglobulin (beta 2-m) amyloid deposition develops serious orthopedic complications in patients with long-term hemodialysis. However, the exact pathogenesis of amyloid deposition in patients with dialysis is unknown. We used transgenic rats with HLA-B27 and beta 2-m to develop an in vivo animal model with beta 2-m amyloid-associated disorders to investigate the mechanism of formation and to report histological findings of beta 2-m amyloid deposition. Transgenic rats were divided into two groups: group 1, no treatment; group 2, arthritis induced by type II collagen. A large number of cells labeled with beta 2-m were observed in the bone marrow and synovium of the knee joint in transgenic rats. In addition, amyloid deposition, identified by Congo red staining, was found only in the knee joints of the transgenic rats with collagen-induced arthritis. Immunostaining with beta 2-m demonstrated the same pattern of tissue distribution as Congo red in serial sections. We hypothesized that elevated serum beta 2-m level-associated local chronic inflammation leads to the development of amyloid deposits and resultant arthropathy.     

人类白细胞抗原-B*2704/B*2705重链拮抗肽的筛选和初步鉴定

目的 从噬菌体展示随机12肽库筛选人类白细胞抗原(HLA)-B·2704及B·2705重链拮抗肽并做初步鉴定.方法 用HLA-B*2704/B*2705重链胞外区蛋白分别筛选噬菌体展爪随机肽库,酶联免疫吸附试验(ELISA)鉴定阳性克隆,DNA测定确定氨基酸序列,免疫荧光和流式细胞术鉴 ,定噬菌体克隆分别与HLA-B*2704及B*2705细胞株结合的特异性.结果 经3轮筛选,获得12个HLA-B·2704拈抗肽,共有5种序列,分别为HTSFCSTHLCLI(×4),QHCSPTLCQIHR(×5),ARCTITL-CYLSN(×1),YGLCTDWYCHIT(×1),YPLCDAILCRLP(×1);10个B*2705拮抗肽共有4种序列,分别为:①SHCSPHWCALPF(×6);②HLCSNSLCLLPW(×2);③EPMCSWFWCTLP(×1);④WTCSPLLCTWGA(×1).比对分析表明,B*2704与B*2705拮抗肽的序列是基本一致的,均含有CS(T)TXXL(W)CXL表位.展示有B*2704拮抗肽的噬菌体克隆可与HLA-B*2704细胞株结合,阳性率为43.55%;而B*2705拈抗肽噬菌体克隆与HLA-B·2705细胞株的阳性结合率为45.69%.结论 筛选获得的HLA-B27拈抗肽的噬菌体克隆具有一定的亲和力,可与表达于细胞株表面的B*2704和B**2705分子特异性结合,而与正常B细胞不结合,因而表现出一定的结合特异性.     

热休克蛋白70-HBcAg(18～27)蛋白复合物对乙型肝炎病毒转基因小鼠的免疫研究

目的研究结核杆菌热休克蛋白(HSP)70-HBcAg(18～27))蛋白复合物对HBV转基因小鼠免疫应答的影响。方法体外构建HSP70-HBcAg(18～27)蛋白复合物,以HBV转基因小鼠为实验对象,进行体内免疫学功能研究。结果HSP70-HBcAg(18～27)蛋白复合物使HBV转基因小鼠外周血及脾中CD4~ T淋巴细胞和CD8~ T淋巴细胞大量增殖,诱导较强的抗原特异性CTL应答,并活化自然杀伤细胞、树突状细胞参与应答反应。结论HSP70-HBcAg(18～27)蛋白复合物具有免疫原性,能够增强转基因鼠细胞免疫应答能力,有望作为蛋白疫苗用于慢性乙型肝炎的免疫治疗。     

Cutaneous polyarteritis nodosa associated with HLA-B39-positive undifferentiated spondyloarthritis in a Japanese patient

Abstract

We present the case of a 43-year-old man diagnosed with HLA-B39-positive spondyloarthritis who developed cutaneous lesions consistent with cutaneous polyarteritis nodosa (CPN). Previous studies indicated an elevated incidence of HLA-B39 in HLA-B27-negative Japanese patients with spondyloarthritis. This case suggested that CPN may also occur in association with forms of HLA-B39-positive spondyloarthritis. The rarity of this association is emphasized. Therapy with corticosteroid and methotrexate improved both the cutaneous lesions and the clinical symptoms of spondyloarthritis.