2-巯基乙醇处理对广州管圆线虫感染大鼠血清特异性IgG检测效果的影响

目的探讨2-巯基乙醇处理对广州管圆线虫感染大鼠血清特异性IgG抗体检测效果的影响。方法用2-巯基乙醇对感染广州管圆线虫大鼠血清处理后,进行ELISA检测,观察标本阳性检出率的变化。结果改进的方法具有较好的敏感性和特异性。通过用2-巯基乙醇对在感染后8、14、20、28、45天和60天6个时点采集的大鼠感染血清进行处理,可不同程度提高其在ELISA反应中的阳性检出率。结论用2-巯基乙醇处理可提高广州管圆线虫感染大鼠血清特异性IgG抗体的阳性检出率。     

严重急性呼吸综合征患者特异性抗体 X线胸片和肺功能的变化

目的通过对严重急性呼吸综合征(SARS)患者出院后血清特异性抗体免疫球蛋白G(IgG)、X线胸片和肺功能的跟踪观察,了解其变化的规律.方法对2003-06～2004-04广州医学院第一附属医院广州呼吸病研究所103例SARS患者出院后定期检查血清特异性抗体IgG、胸片和肺功能,并将其IgG血清浓度与20例健康的医务人员对照比较.结果 (1)SARS患者发病第3天起血清IgG的吸光度(OD)值为(0.063±0.012),与正常对照组(0.016±0.019)比较差异存在显著性(P=0.000),此后逐渐增高,于发病后第3周明显增高达(0.180±0.202),于第4个月达最高值(1.730±0.331),此后逐渐下降,发病后第17个月为(0.668±0.109);(2)62例SARS患者发病后前3个月X线胸片恢复正常的比例为32.3%,发病后第17个月恢复正常的比例为56.5%,异常者有5例,均为重症患者,剔除中断复查的患者(22例),则第17个月胸片正常率为80%;(3)25例SARS患者发病后6个月内肺功能正常者16例,异常者9例,其中4例异常者3～6个月后复查均有不同程度恢复.结论 SARS患者发病后第3天血清IgG的OD值可作为诊断的筛选实验,发病第3周明显增高时可确诊;SARS患者的肺部间质性病变和肺功能损害均可逐渐恢复.     

曼氏裂头蚴可溶性抗原DIGFA法快速检测特异性抗体的潜在诊断价值

目的 探索曼氏裂头蚴可溶性抗原金标免疫渗滤法快速检测曼氏裂头蚴特异性抗体的免疫诊断价值。方法 从自然感染的青蛙中分离出曼氏裂头蚴，经口感染小鼠；于感染后第7周剖杀小鼠，收集鼠血清和虫体并计数。以曼氏裂头蚴可溶性抗原（PsA）为探针建立PsA—DIGFA，分别检测曼氏裂头蚴感染小鼠、健康小鼠、健康人群及其他寄生虫感染者血清中特异性抗体，并与常规ELISA法进行平行检测和比较。结果 DIGFA法检测感染小鼠血清中特异性抗体的敏感性为100％（49／49）、特异性为100％（30／30），与ELISA法检测结果相同；不同虫荷数感染鼠间抗体水平差异无显著性（P〉0．05），感染鼠与正常鼠之间抗体水平差异有显著性（P〈0．01）。PSA—DIGFA检测健康人群血清100人份均为阴性，检测囊虫、包虫、旋毛虫、肺吸虫、华支睾吸虫病病人血清的阳性率分别为42．5％（17／40）、10．0％（2／20）、20．0％（5／25），80．0％（8／10）、8．0％（2／25）。结论 用曼氏裂头蚴粗抗原检测感染动物血清中特异性抗体具有较好的敏感性和特异性，但与其他蠕虫感染存在较高的交叉反应。不同虫荷数感染鼠均产生较高水平抗体，组间差异无显著性。DIGFA法与ELISA法检测的敏感性和特异性相同，但前者更为简便、快速，无需特殊仪器设备，可单份检测，适合寄生虫病的临床检验和流行病学调查。若能进一步纯化检测用抗原，减少与其他蠕虫间的交叉反应，有潜在的应用价值。     

GAGE-7自身抗体在食管鳞状细胞癌中的表达

目的:检测癌-睾丸抗原GAGE-7的自身抗体在食管鳞状细胞癌患者和正常人血清中表达的差异,评价GAGE家族作为肿瘤诊断标志物的可行性.方法:在原核细菌中表达并纯化重组GAGE-7蛋白.采用ELISA检测GAGE-7自身抗体在105份食管鳞状细胞癌患者血清和86份正常人血清中的表达水平.同时检测各血清中总IgG的表达量,计算GAGE-7自身抗体在总IgG中所占比例的差异.结果:血清中GAGE-7抗体在食管鳞状细胞癌中的含量与正常组比较差异显著(P<0.001),而GAGE-7抗体与总IgG比例,患者组明显高于正常组,逻辑回归模型分析c值为0.717,敏感性和特异性分别达到0.72和0.62,且差异显著(P<0.0001).结论:癌-睾丸抗原GAGE-7的自身抗体在食管鳞状细胞癌血清中有较高表达,且与正常组比较有显著差异,其表达特性可被看做是较为理想的癌症检测标志物.     

两种戊型肝炎IgG抗体检测试剂的比较

目的比较2种国产戊型肝炎IgG抗体诊断试剂(E2-IgG和X-IgG)的特异性与敏感度。方法利用2种试剂对2006年江苏省某市普通人群的流行病调查血清标本460份和经过巢式逆转录聚合酶链式反应(nest-RT-PCR)鉴定为HEV核酸阳性的临床戊型肝炎患者血清标本72份进行平行检测,并以免疫印迹实验(Westernblot,WB)和中和单抗阻断试验进行验证。结果1.460份流行病调查血清标本中WB阳性188份。2.188份WB阳性标本中E2-IgG的检测敏感度为99.5,特异度为99.6,阳性预测值99.5,阴性预测值99.6,准确性99.6;X-IgG的检测敏感度为21.3,特异度为97.4,阳性预测值85.1,阴性预测值64.1,准确性66.3。3.E2-IgG阴、阳性A450/A620值的分布在临界值附近分界明显,小于临界值的A450/A620值取对数后呈正态分布,而阳性血清A450/A620值的高低与其WB的反应强度正相关。4.E2-IgG阳性、X-IgG阴性的血清共148份,中和单抗阻断阳性率为98,WB阳性率99.3。5.经PCR鉴定为HEV核酸阳性的急性戊型肝炎患者血清中E2-IgG敏感度98.6,X-IgG敏感度80.6(χ2=10.7,P<0.01)。结论E2-IgG在不同人群中均能较好的反映体内戊型肝炎抗体存在的真实情况。     

细粒棘球蚴感染中国美利奴羊免疫指标的动态变化

目的检测分析绵羊感染细粒棘球蚴免疫指标的变化。方法用细粒棘球蚴对14只健康的中国美利奴羊在相同的条件下进行人工感染,在感染前7d(-7d)、感染当天(0d)、感染后7d、21d、30d、60d用流式细胞仪分析外周血中CD4+T、CD8+T、B淋巴细胞亚群的动态变化,同时检测血清中IgG、IgM、IgE、IFN-γ水平和粒细胞数量。结果进行人工感染的14只绵羊中有8只感染(感染组),6只未感染(未感染组);CD4+T细胞在攻虫后第7d、30d未感染组显著高于感染组(P<0.05)。CD8+T细胞和B淋巴细胞两组之间差异不显著(P>0.05);IgG和IgE水平在攻虫后第30d未感染组显著高于感染组(P<0.05)。IFN-γ水平在攻虫后第21d、30d未感染组显著高于感染组(P<0.05);淋巴细胞在攻虫后第21d(P<0.01)、30d(P<0.05)未感染组显著高于感染组。嗜中性粒细胞在攻虫后第21d感染组极显著高于未感染组(P<0.01)。结论中国美利奴羊感染细粒棘球绦蚴过程中,CD4+T细胞、IgG、IgE、IFN-γ和淋巴细胞水平升高可增强机体对细粒棘球蚴的抵抗力。     

A comparison of the diagnostic efficacy in type 1 autoimmune pancreatitis based on biopsy specimens from various organs

BackgroundComprehensive immunostaining evaluation of the biopsy specimens from various organs with type 1 autoimmune pancreatitis (AIP) has not been elucidated. Our aim was to clarify which of these biopsy specimens and counting method could be a useful tool for supporting the diagnosis of AIP.MethodsWe retrospectively evaluated biopsy specimens from pancreas (n = 19), stomach (n = 28), duodenum (n = 27), duodenal papilla (n = 25), colon (n = 19), liver (n = 11), bile duct (n = 24), and minor salivary gland (n = 13) in 36 patients with AIP. Positive IgG4 immunostaining (>10 plasma cells/high-power field [HPF]) and positive IgG4/IgG ratio (>40%) of biopsy specimens from 8 sites of 6 organs in one HPF and an average from 3 HPFs were compared between AIP and controls.ResultsThe sensitivity of IgG4 immunostaining for AIP in one HPF were 16% in pancreas, 14% in stomach, 15% in duodenum, 52% in duodenal papilla, 11% in colon, 27% in liver, 21% in bile duct and 8% in minor salivary gland, respectively. The positive IgG4 immunostaining of the duodenal papilla in one HPF showed the highest sensitivity (52%) and accuracy (73%) among the 8 sites. It also showed the highest sensitivity among 4 different counting methods (IgG4 immunostaining in one HPF and 3 HPFs, both IgG4 immunostaining and IgG/IgG4 ratio in one HPF and 3 HPFs), but there were no significant differences with respect to specificity and accuracy.ConclusionsIgG4 immunostaining of swollen duodenal papilla with more than 10 IgG4-positive plasma cells in at least one HPF is useful for supporting the diagnosis of AIP.     

肺吸虫特异性IgG4快速检测方法的建立与应用

目的创建简易、快速、准确的肺吸虫病免疫诊断方法。方法用卫氏并殖吸虫成虫粗提液为抗原,以胶体金标记抗人IgG4单抗为检测探针,采用自行设计的垂直流渗滤装置,创建快速检测人血清肺吸虫特异性IgG4方法(P-IgG4-DIGFA),平行检测IgG作比较分析;以经典ELISA为对照,评价其诊断价值,应用P-IgG4-DIGFA检测有效治疗后病人血清,评价其疗效考核价值。结果P-IgG4-DIGFA检测人血清中肺吸虫特异性IgG4的敏感性和特异性分别为95.2%(40/42)、100%(50/50),与血吸虫病患者血清的交叉反应率仅为2%(1/50),未发现与华支睾吸虫病有交叉反应。P-IgG4-DIGFA的敏感性和特异性略高于经典ELISA,差异无统计学意义(χ2=0.25,P>0.05);应用P-IgG4-DIGFA检测有效治疗后3个月、6个月和12个月病人血清特异性IgG4,阴转率分别为0%、16.7%(1/6)和100%。结论金标渗滤法快速检测肺吸虫特异性IgG4,不仅操作简便、快速,而且敏感性高、特异性强,适用于临床检验和现场查病,对治疗后1年病人具有疗效考核价值,有待进一步开发应用。     

无锡市弓形虫感染情况调查

目的 探讨弓形虫感染与特殊人群的关系。方法 以饲养员、屠宰工人、肉熟制品工人、兽医、孕妇、献血员、肿瘤病人等共450人为实验组,另外选择450人为普通人群对照组(样本量、年龄、性别、居住地分别与实验组对应),采用酶联免疫吸附试验对这些人群进行抗弓形虫抗体IgG检测。结果 实验组弓形虫感染IgG阳性率为7.6％,高于对照组(4.0％),两者之间差异有显著性(P<0.05);在实验组各类人群中,饲养员、兽医、献血员、肿瘤病人的感染率最高(均为12％)。结论 无锡市特殊人群弓形虫感染率高于普通人群。     

IgG性质自身抗体导致疑难配血1例

输血工作现在已越来越受到社会各界的重视,而输血前的交叉配血试验则是确保患者安全输血的最后一道非常重要的关卡.笔者在工作中检测出不规则IgG性质的抗自身抗体,干扰了正常配血,现报告如下.     

循环抗原和特异性IgG4对脑囊尾蚴病诊断和疗效评估价值比较

目的比较循环抗原（circulating antigen, CAg ）和特异性IgG4在脑囊尾蚴病诊断和生存状态、 疗效评估中的价值。方法依据CT和MRI的影像表现将68例脑囊尾蚴病患者分为活虫期、变性死亡早期、变 性死亡后期及钙化期四个病期组,检测各病期患者血清中囊虫CAg和IgG4;分别对38例活虫期患者治疗前 ,治疗中,治疗后血清中CAg和特异性IgG4进行检测和比较分析。结果不同病期患者血清CAg阳性率、IgG4 阳性率差异均有显著意义。38例活虫期患者血清CAg、IgG4阳性率分别为100%和98%,随疗程进展,CAg、 IgG4阳性率逐渐降低,不同疗程间CAg阳性率差别有显著性意义,而IgG4阳性率差异无统计学意义。结论 CAg和特异性IgG4能较客观地反映人体内脑囊尾蚴的生存状态,可作为早期诊断指标,两者的特异性及敏 感性无显著差异;对远期疗效评估可以优先选用CAg。     

Possible participation of IgG4 in the activation of complement in IgG4-related disease with hypocomplementemia

Objective: To investigate which IgG subclasses contribute to the activation of the complement pathway in IgG4-related disease (IgG4RD) patients with hypocomplementemia.

Methods: Sera of IgG4RD patients were analyzed for the binding ability of IgG subclasses to complement component 1q (C1q). Polyethylene glycol (PEG) precipitates containing immune complexes (ICs) in sera of IgG4RD patients were analyzed for IgG subclass composition by Western blotting. PEG precipitates containing ICs (PEG-ICs) in sera of patients were also analyzed for their ability to consume complement in normal human serum (NHS) using a total complement hemolytic (CH50) assay and a commercial kit to measure the complement capacity of all three individual complement pathways.

Results: The C1q binding assay revealed high serum levels of C1q-binding IgG4 in IgG4RD patients with hypocomplementemia. ICs in PEG precipitates were formed with IgG4 in IgG4RD patients, regardless of the presence or absence of hypocomplementemia. We observed a marked reduction of CH50 and reduced complement activity in the classical complement pathway as well as the mannan-binding lectin complement pathway in NHS incubated with PEG-IC isolated from IgG4RD patients with hypocomplementemia.

Conclusion: Our results suggest that IgG4 may participate in the activation of complement in IgG4RD patients with hypocomplementemia.     

Some characteristics of aggregates of IgG and plasma proteins in heat-treated factor VIII concentrates

Summary Eight batches of commercial heattreated and one untreated factor VIII concentrate from 6 producers were analyzed for their content of IgG, IgG subclasses, IgG aggregates and the presence of other plasma proteins combined with the IgG as well as for anticomplement activity. Methods used were thin-layer gel filtration, immuno-gel filtration, spot immuno-precipitate assay in a double antibody version and an agarose plate haemolysis inhibition assay of complement fixation. The IgG content varied from 0.1–6.90 g/l. In all preparations IgG existed as monomers and aggregates. Associated with the IgG were also found, at significantly increased amounts compared to normal serum and intravenous immunoglobulin, one to four of the following plasma proteins; fibronectin, fibrinogen, von Willebrand factor antigen, Clq, albumin and IgA. Three batches from two producers had high anticomplementary activity, presumably caused by the IgG aggregates. Two of these deviated strikingly from normal human serum pools in percent distribution of IgG subclasses. It is hypothesized that these aggregates can induce side effects or cause immunological aberrations.     

Graves眼病IgG与干扰素-γ对人球后成纤维细胞ICAM-1、HLA-DR表达的影响

目的　研究Graves眼病 (Graves’ophthalmopathy ,GO)患者血清免疫球蛋白G(IgG)和细胞因子干扰素 γ(IFN γ)对球后成纤维细胞 (retroocularfibroblasts ,RFs)表达细胞间粘附分子 1(ICAM 1)和人类HLA DR的影响 ,探讨细胞免疫和体液免疫在Graves眼病发病中的作用。方法应用 10例GO、7例Graves病、4例结节性甲状腺肿伴甲状腺功能亢进症病人和 8名正常人血清IgG、细胞因子IFN γ及TSH与培养的正常人球后成纤维细胞 (RFs)在体外孵育 ,直接免疫荧光染色后以流式细胞仪检测RFs表达HLA DR、ICAM 1的阳性百分比和平均荧光强度。结果　正常人RFs自发少量表达ICAM 1,不表达HLA DR。GO和其他甲状腺疾病及正常人血清IgG均不能影响ICAM 1和HLA DR的表达。IFN γ、TSH均以剂量依赖方式显著增加RFs表达ICAM 1的阳性百分比和增强平均荧光强度 (P <0 .0 5 ) ,TSH的作用弱于IFN γ的刺激作用。只有IFN γ能刺激RFs表达HLA DR ,并呈剂量依赖关系增加阳性百分比 (P <0 .0 5 )。结论　GO患者血清IgG对培养的正常人球后成纤维细胞表面HLA DR和ICAM 1表达无刺激作用 ;细胞因子IFN γ可能是启动或加重GO自身免疫反应的重要因素 ;TSH可能在GO发病中起一定促进作用。     

Clinical analysis of high serum IgE in autoimmune pancreatitis

AIM: To clarify the clinical significance of high serum IgE in autoimmune pancreatitis (AIP). METHODS: Forty-two AIP patients, whose IgE was measured before steroid treatment, were analyzed. To evaluate the relationship between IgE levels and the disease activity of AIP, we examined (1) Frequency of high IgE (> 170 IU/mL) and concomitant allergic dis-eases requiring treatment; (2) Correlations between IgG, IgG4, and IgE; (3) Relationship between the presence of extrapancreatic lesions and IgE; (4) Re-lation...     

唾液幽门螺杆菌IgG测定在胃癌初步筛查中的价值

胃癌的症状常无特异性,大多数患者诊断时己属中晚期,预后较差.但若能得到早期诊断和及时治疗,则预后甚好,5年存活率在80%以上.     

Size- and charge selectivity of glomerular filtration in Type 1 (insulin-dependent) diabetic patients with and without albuminuria

Summary Albuminuria is the first clinical event in the development of diabetic nephropathy. We assessed glomerular charge- and size selectivity in 51 patients with Type 1 (insulin-dependent) diabetes mellitus of juvenile onset and 11 healthy individuals. Patients were allocated to five groups. The urinary albumin excretion rate was normal in group D1; 30–100 mg/24 h in group D2; 101–300 mg/24 h in group D3 and greater than 300 mg/24 h in groups D4 and D5. Group D5 had elevated serum creatinine (above 110 mol/l). Glomerular filtration rate and renal plasma flow were determined by constant infusion techniques and tubular protein reabsorption by excretion of 2-microglobulin. Charge selectivity was estimated from the IgG/IgG4 selectivity index. Size selectivity was measured by dextran clearance. Dextran was measured by refractive index detection after fractionation (2 Å fractions in the range 26–64 Å) by size exclusion chromatography. IgG/IgG4 selectivity index was significantly decreased in patients with albuminuria (p<0.001). The drop in IgG/IgG4 selectivity index was found in patients with minimal albuminuria (D2) and was not accompanied by any changes in tubular function or glomerular haemodynamics. Size selectivity was significantly altered only in patients with the most advanced nephropathy (D5) as reflected by an increase in the clearance of 62 Å dextran (p<0.04). We conclude that loss of glomerular charge selectivity precedes or accompanies the formation of new glomerular macromolecular pathways in the development of diabetic nephropathy.     

Review of primary sclerosing cholangitis with increased IgG4 levels

Primary sclerosing cholangitis(PSC) is a chronic progressive liver disease. Subtypes of PSC have been described, most recently PSC with elevated serum and/or tissue IgG4 subclass. We aim to summarise the clinical phenotype,disease associations, differential diagnosis, response to therapy and pathogenic mechanisms underlying PSC-high IgG4 subtype. We reviewed Pub Med,MEDLINE and Embase with the search terms "primary sclerosing cholangitis","IgG4", and "IgG4-related sclerosing cholangitis(IgG4-SC)". Elevated serum IgG4 are found in up-to one-quarter, and abundant IgG4-plasma cell infiltrates in the liver and bile ducts are found in up-to one-fifth of PSC patients. This group have a distinct clinical phenotype, with some studies reporting a more aggressive course of liver and associated inflammatory bowel disease, compared to PSCnormal IgG4 and the disease mimic IgG4-SC. Distinguishing PSC-high IgG4 from IgG4-SC remains challenging, requiring careful assessment of clinical features,organ involvement and tissue morphology. Calculation of serum IgG4:IgG1 ratios and use of a novel IgG4:IgG RNA ratio have been reported to have excellent specificity to distinguish IgG4-SC and PSC-high IgG4 but require validation in larger cohorts. A role for corticosteroid therapy in PSC-high IgG4 remains unanswered, with concerns of increased toxicity and lack of outcome data. The immunological drivers underlying prominent IgG4 antibodies in PSC are incompletely defined. An association with PSC-high IgG4 and HLA class-II haplotypes(B*07, DRB1*15), T-helper2 and T-regulatory cytokines(IL4, IL10,IL13) and chemokines(CCL1, CCR8) have been described. PSC-high IgG4 have a distinct clinical phenotype and need careful discrimination from IgG4-SC,although response to immunosuppressive treatments and long-term outcome remains unresolved. The presence of IgG4 likely represents chronic activation to persistent antigenic exposure in genetically predisposed individuals.     

血清免疫球蛋白(Ig)G4在非IgG4相关性肝胆疾病中的表达

回顾性分析因腹痛、黄疸、肝功能异常等肝胆系统相关症状而就诊并进行血清免疫球蛋白(Ig)G4检测的患者,探讨血清IgG4在非IgG4相关性肝胆疾病患者中的表达情况及其临床意义,发现血清IgG4水平升高亦见于非IgG4相关性肝胆疾病(IgG4-RD)患者,性别和年龄可能对血清IgG4水平造成一定的影响。通过随访IgG4-RD患者的血清IgG4水平和/或病理组织学检查有助于提高对IgG4-RD以及血清IgG4水平价值的认识。