HLA-B27 polymorphism in patients with juvenile and adult-onset ankylosing spondylitis in Southern China

Abstract
Distribution of B27 subtypes in juvenile and adult-onset ankylosing spondylitis (JAS and AAS) in Southern China was studied. A total of 505 patients belonged to Han population were included (145 JAS and 360 AAS patients), and 1368 healthy individuals were included as controls. Human leukocyte antigen (HLA)-B27 typing was performed by Luminex liquid array combining polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) and/or serological method. HLA-B27 subtyping was performed by polymerase chain reaction-sequence specific primer (PCR-SSP). The sequence-based typing was performed for the B*2715 samples to verify the PCR-SSP results. HLA-B27 was presented in 453 of 505 patients (89.7%), compared with 74 of 1368 controls (5.41%). B*2704 subtype in AS group was significantly higher than controls and B*2705 subtype significantly lower. B*2715 and B*2702 were found in 1.32% and 0.66% of the B27-positive patients but none in controls, and there was no significant difference between either of them and controls. B27-positive patients were 134 (92.4%) in JAS group and 319 (88.6%) in AAS group. There was no significant difference for B27 subtypes distribution between JAS (B*2704, 05, 15) and AAS (B*2704, 05, 15, 02) groups. The frequency of B*2715 in two groups was 3 (2.24%) and 3 (0.94%), respectively. The onset age of three JAS patients carrying B*2715 was 5, 9 and 13 years old, respectively. Our results suggested that B*2704 was the predominant subtype in AS patients in Southern China. B*2715 was observed in AS group only and slightly more in JAS than in AAS, and the patients carrying this allele tended to have early onset, B*2715 may be disease-association subtype.     

停用依那西普后强直性脊柱炎病情复发相关因素分析及骨灵汤对其病情复发的影响

目的探讨停用依那西普后强直性脊柱炎（AS）病情复发情况及其相关因素,评价骨灵汤对病情复发的影响。方法 60例活动性AS随机分为两组,治疗1组（骨灵汤合依那西普组）30例,治疗2组（依那西普组）30例,基线均合用一种非甾体类抗炎药。治疗12周后达到ASAS20改善的患者,停用依那西普,基线药物继续使用,每2～4周定期随访,直至病情复发或至9个月。采用Cox比例风险模型评价与复发有关的因素,Log-rank检验比较两组之间的复发情况。结果 12周后共53例患者达到了ASAS20改善标准并进入随访研究。其中治疗1组28例,治疗2组25例,两组人口统计学资料、基线及12周时病情特征比较差异无统计学意义（P〉0.05）。至研究结束时,有81.1%的患者病情复发。治疗1组复发率为71.4%,复发时间中位数为20周;治疗2组复发率为92.0%,复发时间中位数为14周,两组差异有统计学意义（P〈0.05）。Cox回归模型分析显示,基线BASDAI和CRP高及髋关节受累是病情复发的危险因素（P〈0.05）,治疗方法为保护因素（P〈0.05）。结论 AS停用依那西普后维持原药物治疗多数患者病情会复发,基线高BASDAI、CRP和髋关节受累是疾病复发的不利因素。早期联合使用骨灵汤能够有效维持疗效,延缓复发。     

强直性脊柱炎患者MICA基因第2、3和4外显子的多态性及其与HLA-B抗原的连锁不平衡

目的探讨皖籍汉族人群MICA基因（major histocompatibility complex class Ⅰchain-related gene A，MICA）第2、3、4外显子的多态性，及其与HLA-B抗原的连锁不平衡在强直性脊柱炎（ankylosing spondylitis，AS）发病中的作用。方法采用聚合酶链反应-序列特异性寡核苷酸探针杂交（polymerase chain reactionsequence-specific oligonucleotide probing，PCR-SS0）技术对56例AS患者和112名正常对照人群进行MICA基因第2、3、4外显子的多态性和HLA-B抗原的检测。结果AS患者和正常对照人群的MICA等位基因分布均以MICA＊008占优势，频率分别为32．14％和30．36％。两组人群MICA＊007等位基因的分布差异有统计学意义（X^2=10．18，P〈0．05，RR=2．50）。单倍型分析显示，AS患者和正常对照人群的MICA等位基因均显示出与多个HLA-B位点的连锁不平衡现象，两组间差异有统计学意义的单倍型为MICA＊007-B27（X^2=18．46，P〈0．05，RR=7．47）。分层分析结果显示，HLA-B27阳性与AS的相关性有统计学意义（P〈0．05），但MICA＊007基因与AS的相关性无统计学意义（P〉0．05）。结论AS患者中MICA＊007等位基因频率的显著升高可能源于MICA基因与HLA-B位点间的广泛连锁不平衡。     

Successful etanercept therapy for refractory sacroiliitis in a patient with ankylosing spondylitis and mixed connective tissue disease

The concurrence of ankylosing spondylitis (AS) in a patient with mixed connective tissue disease (MCTD) is rarely described in the literature. Significant and sustained efficacy with tumor necrosis factor (TNF)-alpha blockers has been demonstrated in AS patients. However, evidence to date has revealed associated side effects, including antinuclear antibody induction and development of a lupus-like syndrome. Several authors have reported lupus-like manifestations in MCTD patients treated with TNF-alpha blockers used to control peripheral polyarthritis. In our case report, we demonstrate a good response to etanercept therapy for refractory sacroiliitis in a patient with coexisting AS and MCTD, without development of a lupus-like syndrome. This demonstrates that etanercept therapy may be an appropriate therapeutic agent for sacroiliitis in MCTD patients, as it is in AS alone.     

High levels of soluble GPR56/ADGRG1 are associated with positive rheumatoid factor and elevated tumor necrosis factor in patients with rheumatoid arthritis

Background

GPR56/ADGRG1 is a member of the adhesion-class G protein-coupled receptor (aGPCR) family important in brain development, oncogenesis and tumor metastasis. Like other aGPCRs, GPR56 is cleaved at the GPCR proteolysis site (GPS) motif into an N-terminal fragment (NTF) and a C-terminal fragment (CTF). Existence of soluble GPR56 (sGPR56) has been shown in vitro, however the underlying mechanism and its pathophysiologic role remains undetermined.

ERAP1 reduces accumulation of aberrant and disulfide-linked forms of HLA-B27 on the cell surface

ObjectiveEndoplasmic reticulum (ER) aminopeptidase 1 (ERAP1) variants contribute to the risk of ankylosing spondylitis in HLA-B27 positive individuals, implying a disease-related interaction between these gene products. The aim of this study was to determine whether reduced ERAP1 expression would alter the cell surface expression of HLA-B27 and the formation of aberrant disulfide-linked forms that have been implicated in the pathogenesis of spondyloarthritis.MethodsERAP1 expression was knocked down in monocytic U937 cells expressing HLA-B27 and endogenous HLA class I. The effect of ERAP1 knockdown on the accumulation HLA-B alleles (B18, B51, and B27) was assessed using immunoprecipitation, isoelectric focusing, and immunoblotting, as well as flow cytometry with antibodies specific for different forms of HLA-B27. Cell surface expression of aberrant disulfide-linked HLA-B27 dimers was assessed by immunoprecipitation and electrophoresis on non-reducing polyacrylamide gels.ResultsERAP1 knockdown increased the accumulation of HLA-B27 on the cell surface including disulfide-linked dimers, but had no effect on levels of HLA-B18 or -B51. Antibodies with unique specificity for HLA-B27 confirmed increased cell surface expression of complexes shown previously to contain long peptides. IFN-γ treatment resulted in striking increases in the expression of disulfide-linked HLA-B27 heavy chains, even in cells with normal ERAP1 expression.ConclusionsOur results suggest that normal levels of ERAP1 reduce the accumulation of aberrant and disulfide-linked forms of HLA-B27 in monocytes, and thus help to maintain the integrity of cell surface HLA-B27 complexes.     

强直性脊柱炎临床病理分期与疼痛缓解程度的临床应用研究

目的通过分析强直性脊柱炎术后临床病理分期,探讨腰背部椎旁肌有限剥离术缓解活动性强直性脊柱炎的疗效与临床病理分期的关系。方法 2007~2014年收集符合强直性脊柱炎诊断标准、资料完整的活动期患者127例,均实施了腰背部椎旁肌有限剥离术,观察术前术后Bath强直性脊柱炎病情活动性指数(BASDAI),Bath强直性脊柱炎功能指数(BASFI),Bath强直性脊柱炎测量指数(BASMI),Bath强直性脊柱炎健康综合指数(BASG)评分,根据病理结合临床行分期,同时观察腰背部椎旁肌有限剥离术治疗强直性脊柱炎疗效与临床病理分期的关系。结果腰背部椎旁肌有限剥离手术无任何并发症、后遗症,腰部疼痛均有不同程度的缓解,BASDAI、BASFI与BAS-G 3项指标术后2周开始明显下降,而BASMI术后与术前相比未见明显差异。临床病理分为三期,临床症状改善情况与临床病理分期呈负相关关系,早期缓解明显,具有显著统计学意义。结论在病情早期实施腰背部椎旁肌有限剥离术能明显缓解活动性强直性脊柱炎临床症状,可有效抑制病情的进一步进展。     

ELIspot法观察中药红曲对强直性脊柱炎外周血单个核细胞分泌TNF-α的影响

目的 使用ELIspot法观察中药红曲对强直性脊柱炎(ankylosing spondylitis)患者外周血单个核细胞分泌TNF-α的影响.方法 采集强直性脊柱炎病情活动期患者(均符合1984年美国风湿病协会关于强直性脊柱炎的诊断)的外周血,分离外周血单个核细胞(peripheral blood mononuclear cells,PBMCs),制成细胞悬液,以每孔2.5 ×105个细胞种植到包被有抗肿瘤坏死因子α(tumor necrosis factor α,TNF-α)抗体的96孔酶联免疫(enzyme-linked immunospot,ELIspot)板上.分为正常对照组、阴性对照组(TNF-α抑制剂组)、阳性对照组[脂多糖(lipopolysaccharide,LPS)刺激]、红曲高浓度组(0.5mg/mL)、红曲中浓度组(0.25mg/mL、红曲低浓度组(0.125 mg/mL),细胞种植的同时给予阴性药物、阳性药物和红曲高中低三种浓度药物刺激,24后检测TNF-α分泌情况.结果 红曲高中低浓度组的ELIspot检测,斑点计数均少于正常组,其中红曲高、中浓度组与正常对照组比较差异有统计学意义(P＜0.05).结论 ELIspot检测是一种较好的用于观察药物对细胞作用效果的方法;红曲能减少强直性脊柱炎患者外周血单个核细胞TNF-α的分泌.