用原位聚合酶链反应方法检测结节病病理组织中博氏疏螺旋体的研究

目的进一步探讨结节病与博氏疏螺旋体（Ｂｂ）感染之间的关系。方法用原位聚合酶链反应（ＰＣＲ）和免疫荧光方法分别对２３例结节病患者病理活组织检查（活检）组织中Ｂｂ鞭毛ＤＮＡ和５５例血清标本中Ｂｂ抗体进行了检测。结果（１）２３例病理组织中未有１例检测到Ｂｂ鞭毛ＤＮＡ的存在。（２）５５例结节病患者血清中Ｂｂ抗体总阳性率为５５％，明显高于正常人的１０％（Ｐ＜０．００５）。结论Ｂｂ可能不是结节病的病因，结节病患者血清Ｂｂ抗体的升高，可能是机体非特异性反应的结果     

抗肺炎克雷白杆菌亚单位抗体与强直性脊柱炎

以免疫印迹法（Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ）测定２６例强直性脊柱炎（ＡＳ）患者和７例类风湿关节炎（ＲＡ）病人及正常人抗肺炎克雷白杆菌（ＫＰ）抗体阳性血清中抗ＫＰ亚单位抗体。结果表明，ＡＳ患者血清抗ＫＰ亚单位抗体阳性区带条数为６．６±１．９条，ＲＡ患者和正常人为３．６±１．３条（Ｐ＜０．０５）。ＡＳ患者血清抗ＫＰ亚单位抗体以６４６００，４８２００以及３６０００抗体占多数，阳性率分别为８０．７％，６１．５％和６５．４％；ＲＡ患者和正常人血清则以抗３６０００和３００００亚单位抗体占多数，阳性率分别为７５％和５０％。抗ＨＬＡ－Ｂ＿（２７）单价血清和兔抗人工合成１８肽（含与ＨＬＡ－Ｂ＿（２７）相同氨基酸序列片段）抗血清能与ＫＰ抗原中的６４６００和４８２００两个亚单位分子起交叉反应，提示这两个亚单位分子与ＨＬＡ－Ｂ＿（２７）有共同的抗原成分，推测ＫＰ感染可能通过与ＨＬＡ－Ｂ＿（２７）的分子模拟在ＡＳ的发病中起作用。     

检测抗核包膜蛋白抗体的临床意义初探

目的进一步探讨抗核包膜蛋白抗体的临床意义。方法对48例原发性胆汁性肝硬化（PBC）及1881例对照组血清标本以间接免疫荧光法检测抗核抗体（ANA），选择抗核包膜蛋白抗体阳性的病例进行临床回顾性分析。结果1929例血清标本中50例抗核包膜蛋白抗体阳性，相关疾病包括PBC等多种系统性自身免疫病。抗核包膜蛋白抗体在PBC患者组中的阳性率为39．58％，明显高于其他系统性自身免疫性疾病组（x^2=134，P〈0．01）及非自身免疫性肝脏疾病组即；109，P〈0．01），诊断PBC的特异性为98．35％。结论抗核包膜蛋白抗体与多种疾病相关，这种现象可能与其靶抗原的异质性相关。但其对诊断PBC价值较高。     

双夹心DIGFA检测血吸虫循环抗原的应用研究

以日本血吸虫重组极低密度脂蛋白（ＳＶＬＢＰ）制备的多抗为捕捉抗体，胶体金标记血吸虫虫卵多克隆抗体为覆盖抗体，采用双夹心金标免疫渗滤法（ＤＩＧＦＡ）检测血吸虫病病人血清中循环抗原。结果７０份急性血吸虫病病人血清的阳性率为１００％，１５９份慢性血吸虫病病人血清的阳性检出率为６２．３％；１２０份流行区健康人群血清检测均阴性；１００份非流行区健康人群血清，２份呈阳性反应，假阳性率为２％。与肺吸虫病病人血清     

肺孢子虫病实验与临床研究 Ⅱ．免疫印迹试验检测人血清肺孢子虫抗体

采用ＳＤＳ－ＰＡＧＥ分析鼠源卡氏肺孢子虫包囊可溶性抗原呈现２０条以上多肽区带，主带分子量分别为＞１００、８５－９４、６７、５２和３４ｋＤａ。ＥＩＴＢ检测表明重庆地区健康人血清中存在识别１１０、１０５、８５、６７、５２和４６ｋＤａ的ＩｇＧ型抗体，抗体阳性率为５６．７％（５９／１０４）。其中８５ｋＤａ抗原能与抗人源肺孢子虫单克隆抗体２Ｇ２起反应。健康人血清抗８５ｋＤａ抗原的抗体阳性率为３７．５％（３９／１０４），但ＩｇＭ型抗体均为阴性。另外检测７例确诊的卡氏肺孢子虫病患者，４例血清ＩｇＧ型抗体阳性，其中３例抗８５ｋＤａＩｇＧ型抗体阳性。     

抗α-胞衬蛋白抗体在干燥综合征诊断及病情判断中的作用

目的探讨抗α-胞衬蛋白（Fodrin）抗体在干燥综合征（SS）诊断及病情判断中的作用。方法应用酶联免疫吸附试验（ELISA）检测40例原发性SS（pSS）、24例继发性SS（sSS）、32例其他结缔组织病（CTD）和31例非CTD患者血清抗α-Fodrin—IgA、IgG抗体,同时记录SS患者的临床资料。结果抗α-Fodrin-IgA、IgG抗体诊断ss的敏感性分别为31．3％、28．1％,特异性分别为90．5％、88．9％;同时检测两型抗体诊断ss的敏感性显著提高,达48．4％（P〈0．05）。pSS和sSS患者抗α-Fodrin抗体（I型或Ⅱ型）的阳性率均显著高于非CTD组（P〈0．05）;SS患者抗α-Fodrin抗体的出现与临床症状、眼部体征及核素检查显示的唾液腺功能受损无明显相关性（P〉0．05）;抗α-Fodrin抗体阳性患者血清C反应蛋白高于阴性者（P〈0．05）;抗核抗体（ANA）、抗SSA（干燥综合征抗原A）、抗SSB（干燥综合征抗原B）抗体3项阴性的患者抗α-Fodrin抗体阳性率为36．4％。结论抗α-Fodrin抗体特异性较好,对诊断SS有一定参考价值,尤其对于ANA、抗SSA、抗SSB抗体阴性的患者,且可能有助于评估SS病情活动性。     

~（99m）Tc－ccM4（鼠／人嵌合抗体）胃癌放射免疫显像研究

采用~（９９ｍ）Ｔｃ－ｃｃＭ４ＭｃＡｂ对１０例胃癌病人，４例非胃癌病人进行放射免应显象（ＲＩＩ）研究，观察了血清ＴＡＧ－７２抗原量、抗体注入量、组织类型和细胞反应性对ＲＩＩ阳性检出率的影响。注入（９９ｍ）Ｔｃ－ｃｃＭ４（５５５～１４８０ＭＢｑ／０．５～１．５ｍｇ）后，胃癌阳性率６０％，肝转移癌阳性率５０％，全部良性病变均显示阴性。最佳显象时间为注入抗体后８ｈ。给予一定量抗体后，增加抗体用量并不能提高ＲＩＩ的阳性率。血清ＴＡＧ－７２抗原含量与ＲＩＩ肿瘤阳性检出率明显相关。     

慢性丙肝RIBA与RIA-2检测抗HCV的价值

目的探讨重组免疫印迹试验（ＲＩＢＡ）在慢性ＨＣＶ感染中的应用价值．方法１９９３年１月～１９９５年３月采用含ＨＣＶ不同编码区三种重组抗原（Ｃ２２，Ｃ３３ｃ，Ｃ１００３）的ＲＩＢＡ，对８５例慢性非甲非乙型肝炎（ＮＡＮＢＨ）患者血清进行检测并将其结果与第二代放免法（ＲＩＡ２）检测抗ＨＣＶ及逆转录多聚酶链反应（ＲＴＰＣＲ）检测ＨＣＶＲＮＡ结果进行比较分析．结果慢性ＮＡＮＢＨ患者血清ＲＩＢＡ阳性（７８８％，６７／８５）结果与ＨＣＶＲＮＡ阳性（７５３％，６４／８５）结果之间有良好相关性和高的符合率；ＲＩＢＡ可检出ＲＩＡ２阴性的ＨＣＶ感染者；单项抗体阳性时仅见一定比例的单项抗Ｃ２２或抗Ｃ３３ｃ阳性血清可检出ＨＣＶＲＮＡ，而单一抗Ｃ１００３阳性者ＨＣＶＲＮＡ均阴性．结论ＲＩＢＡ可提高慢性ＨＣＶ感染的诊断率，ＲＩＢＡ阳性为ＨＣＶ感染及病毒血症存在的标志，可作为ＲＩＡ２检测抗ＨＣＶ的确证实验．     

抗中性粒细胞胞浆抗体在炎性肠病中的意义

目的 研究抗中性粒细胞胞浆抗体（ＡＮＣＡ）在炎性肠病（ＩＢＤ）中的发生率、其针对的靶抗原及其与临床疾病活动性的关系。方法 用间接免疫荧光法对７６例溃疡性结肠炎（ＵＣ）、３６例克隆病（ＣＤ）及２１０例正常者进行ＡＮＣＡ的筛选测定，用ＥＬＩＳＡ检测ＡＮＣＡ针对的不同靶抗原。结果 ７６例ＵＣ患者中血清ＡＮＣＡ阳性者占７１．１％，均为核周型染色，明显高于ＣＤ患者ＡＮＣＡ的阳性率（８．３％，Ｐ〈０．００１）     

抗Sa抗体在类风湿关节炎中的意义

目的测定自身免疫性结缔组织病中抗Ｓａ抗体的阳性率，着重分析抗Ｓａ抗体在类风湿关节炎（ＲＡ）中的意义。方法从人胎盘中提取Ｓａ抗原，采用免疫印迹法，测定了４０例健康人及４７８例各种自身免疫性结缔组织病患者血清中的抗Ｓａ抗体，并分析了该抗体与ＲＡ的某些临床及实验室指标的相关性。结果抗Ｓａ抗体在各组患者中的阳性率分别为：ＲＡ为３１．９％（６１／１９１），干燥综合征为３．０％（２／６７），系统性红斑狼疮为４．３％（２／４６），白塞病、肌炎／皮肌炎、其他自身免疫性结缔组织病及正常人中均为０。研究分析表明，抗Ｓａ抗体对ＲＡ的诊断敏感性为３１．９％，特异性为９８７％，阳性预报率为９３．８％，阴性预报率为７１．３％。与抗Ｓａ抗体阴性的ＲＡ患者相比，抗Ｓａ抗体阳性的患者在关节受累、晨僵、血沉、抗核抗体、Ｘ线分期和二线药物的使用方面有显著差异。结论我们在国内首次制备了Ｓａ抗原，并建立了Ｓａ抗体的检测方法，它是一种不同于类风湿因子、对ＲＡ诊断较为特异的新型自身抗体。该抗体与疾病的严重程度相关，能否有助于ＲＡ的早期诊断和分型尚需更多病例的积累和观察。     

Elevated IgG4 levels in patients demonstrating sarcoidosis-like radiologic findings

One of the radiologic patterns associated with IgG4-related systemic disease was similar to that of pulmonary sarcoidosis. We analyzed whether suspected pulmonary sarcoidosis might include unrecognized IgG4-related systemic disease. The enrolled patients had bilateral hilar lymphadenopathy and/or lung nodules on chest computed tomography, used to diagnose the patients who could either be compatible with or suggested as having pulmonary sarcoidosis. The IgG4 levels were retrospectively measured. Bronchoalveolar lavage (BAL) was analyzed for the presence of IgG subclasses, and specimens were stained by an antibody to IgG4. We compared these data in the suspected sarcoidosis patients, with or without elevated serum IgG4, with the laboratory data and bronchoscopy results in patients with definite sarcoidosis. All enrolled patients were followed for over 5 years. The patients were classified as 49 definite and 44 suspected sarcoidosis patients. Eight patients, including 6 suspected sarcoidosis patients, had elevated abnormal levels of serum IgG4. The suspected sarcoidosis patients had significantly lower percentages of lymphocytes and IgG in the BAL. One suspected sarcoidosis patient had positive IgG4 staining in a lung specimen. The elevated serum IgG4 patients among the patients with suspected sarcoidosis showed significantly higher levels of BAL IgG4, IgG4/IgG, and IgG4/IgG3 compared with the levels of the normal serum IgG4 patients. The follow-up study revealed that 1 patient with elevated serum IgG4 was complicated with other organ failure caused by IgG4-related systemic disease, and Castleman disease was diagnosed in 2 patients. IgG4-related systemic disease was, therefore, identified among the patients with elevated serum IgG4.     

Hyperprolactinemia in sarcoidosis: incidence and utility in predicting hypothalamic involvement.

Patients with sarcoidosis have been reported frequently to have elevated concentrations of serum prolactin. On this basis, it was suggested that the hypothalamus might be a common site of involvement by sarcoidosis and that measurement of serum prolactin concentrations might serve as a sensitive indicator of hypothalamic disease. We measured serum prolactin concentrations in a group of 61 patients with sarcoidosis. Hyperprolactinemia was detected in only 2 of the entire group and was not observed in any of the 9 patients with central nervous system involvement. Because radioimmunoassayable prolactin concentrations are infrequently elevated in patients with disseminated sarcoidosis, even when pitutitary hypofunction is apparent, it is concluded that the measurement of serum prolactin is not a reliable method for screening these patients for pituitary or hypothalamic disease.     

Elevated Tenascin-C Levels in Bronchoalveolar Lavage Fluid of Patients with Sarcoidosis

Background

Sarcoidosis is a disease characterized by granulomatous lesions involving multiple organ systems. The etiology of sarcoidosis remains unknown, and reliable biomarkers have not been identified. Tenascin-C is an extracellular matrix molecule expressed during wound healing in various tissues. The present study aimed to investigate the role of tenascin-C in sarcoidosis.

Methods

Enzyme-linked immunosorbent assays were used to measure tenascin-C levels in serum and bronchoalveolar lavage fluid (BALF) from 31 patients with sarcoidosis and 15 healthy individuals. Relationships between tenascin-C concentrations in BALF and serum samples and clinical parameters in patients with sarcoidosis were evaluated.

Results

BALF tenascin-C levels were significantly higher in patients with sarcoidosis than in healthy individuals, but serum levels were no different. BALF tenascin-C levels showed positive correlations with serum lactic dehydrogenase levels and the ratio of lymphocytes in BALF. BALF tenascin-C levels were also higher in patients with parenchymal infiltration on chest radiographs than in those without.

Conclusions

The present results demonstrated that the BALF tenascin-C level was correlated with pulmonary infiltrates on chest radiographs in patients with sarcoidosis. Although measurement of serum tenascin-C levels has a limited role and measurement of BALF tenascin-C levels might be impractical, tenascin-C in the lung might play a role in the pathogenesis of sarcoidosis. Further studies are necessary to determine the role of BALF tenascin-C in sarcoidosis.     

Borrelia burgdorferi may be the causal agent of sarcoidosis]

Serum antibody to Borrelia burgdorferi was measured in 33 patients with sarcoidosis who were confirmed clinically and pathologically. The results showed that 81.8% of the patients were positive. In addition, a strain of Borrelia burgdorferi was isolated from a patient's blood. Fourteen patients received ceftriaxone 2 g per day and/or penicillin 12 million per day and a patient received lincomycin 1.2 g per day. The antibody titer of the patients turned to normal level, their SACE turned to normal range, and chest X-ray findings were markedly improved in 3 cases after the treatment. According to the facts mentioned above, we consider that Borrelia burgdorferi may be the causal agent of sarcoidosis and sarcoidosis might be a special type of Lyme disease.     

Vitamin D receptor gene polymorphism in patients with sarcoidosis.

The active form of vitamin D, 1,25-dihydroxyvitamin D(3), is known to be produced at sites of granulomatous reactions in sarcoidosis. 1, 25-dihydroxyvitamin D(3) has multiple immunomodulatory effects, and acts as a promoter of multinucleated giant cell formation. Polymorphism of the vitamin D receptor (VDR) gene has recently been shown to be related to bone mineral density, and also associated with hyperparathyroidism and risk of prostatic carcinoma. Considering that this might affect sarcoidosis, we investigated polymorphism of the VDR gene in 101 patients with sarcoidosis and 105 healthy control subjects. Their genotypes were determined using polymerase chain reaction (PCR) and restriction fragment length polymorphism. In the patients with sarcoidosis, the BB, Bb, and bb genotypes accounted for 1.0%, 37.6%, and 61.4%, whereas in healthy control subjects the figures were 1.0%, 20.0%, and 79.0%, respectively. The difference in the genotype distribution between healthy control subjects and sarcoidosis patients was significant (p < 0.05) with the frequency of the B allele being elevated (p < 0.05). From the result, we suggest that in VDR gene polymorphism the B allele might be a genetic risk factor for sarcoidosis.     

Angiotensin Converting Enzyme in Sarcoidosis and in Silicosis

Angiotensin converting enzyme(ACE) activities of broncho-alveolar lavage fluid(BALF) and serum in patients with sarcoidosis and with silicosis were measured. Serum ACE was measured by enzymic assay and radioimmunoassay. There was a close relationship between ACE activity and content (r=0.78). Serum ACE activities in patients with active sarcoidosis (37.5 ± 11.1 nmol/min/ml, mean ± SD) and with silicosis (25.5 ± 9.3) were significantly elevated from the control (18.6 ± 6.0). BALF ACE activities in the control, patients with active sarcoidosis and with silicosis were 0.23 ± 0.19 nmol/min/ml, 0.94 ± 0.97 and 0.38 ± 0.05, respectively. BALF ACE in patients with active sarcoidosis and with silicosis were significantly different from the control. When BALF ACE was corrected by the cell count of alveolar macrophage (per 10⁶ cells), activity was significantly different from control only in the patients with sarcoidosis. Moreover, only the alveolar macrophages in sarcoidosis were stained by immunofluorescence and immunocytochemistry using rabbit anti-human ACE antibody. Induction of ACE in alveolar macrophage might have an important role for the activity or progression of sarcoidosis.     

Chitotriosidase activity in the serum of patients with sarcoidosis and pulmonary tuberculosis

BACKGROUND: Human chitotriosidase is a chitinase selectively expressed by activated macrophages. An increase in chitotriosidase activity was previously described by us in the serum and bronchoalveolar lavage of sarcoidosis patients. OBJECTIVE: The aim of the present study was to analyze serum chitotriosidase activity in a larger number of sarcoidosis patients to verify the reported increase with respect to controls and to compare serum chitotriosidase levels in patients with sarcoidosis and tuberculosis, two granulomatous disorders of different etiology. METHODS: Chitotriosidase activity was measured in the serum of 96 sarcoidosis patients, 15 pulmonary tuberculosis patients and 30 healthy controls. RESULTS: We found significantly higher serum chitotriosidase activity in sarcoidosis patients than controls (p < 0.01) and in sarcoidosis patients than tuberculosis patients (p < 0.01), confirming a striking elevation of chitotriosidase activity (>10 times greater than normal) in pulmonary sarcoidosis patients. This is the first time that chitotriosidase activity has been analyzed in the serum of patients with pulmonary tuberculosis; it was found to be significantly lower than in sarcoidosis patients and not significantly greater than in controls. CONCLUSION: Although the mechanisms leading to the increase in chitotriosidase activity in sarcoidosis are still unknown, this enzyme may be specifically involved in the pathogenesis of the disease. Further studies with a greater number of patients are needed to confirm these results and to determine whether chitotriosidase could be a marker with diagnostic or prognostic value in sarcoidosis.     

Elevated level of soluble HLA class I antigens in serum and bronchoalveolar lavage fluid in patients with sarcoidosis

OBJECTIVE: Soluble HLA class I antigens (sHLAs) in human serum have been reported to be associated with allografts and autoimmune disease and could modify immunological reactions induced by membrane type HLAs. To investigate the clinical significance of sHLAs in sarcoidosis, we assessed concentrations of sHLAs in both serum and bronchoalveolar lavage fluid (BALF) and also examined their production by peripheral blood mononuclear cells (PBMCs) and BALF cells. METHODS: Concentrations of sHLAs were determined by enzyme-linked immunosorbent assay, using a monoclonal antibody against HLA class I (W6/32) and an enzyme-labeled polyclonal antibody to human beta2-microglobulin. PBMCs and BALF cells were cultured in the presence or absence of either LPS or PHA. PATIENTS: Serum levels of sHLAs were assessed in 96 patients with sarcoidosis and in 32 healthy control subjects. sHLAs concentrations in BALF were also investigated in 17 active sarcoidosis patients and in 13 control subjects. RESULTS: sHLAs levels in both serum and BALF were higher in sarcoidosis cases than in control subjects (p<0.05, in both). In the patients, values were significantly higher in active than in inactive stages (p<0.001) and significantly correlated with angiotensin-converting enzyme (ACE) levels. Both PBMCs and BALF cells produced enhanced amounts of sHLAs in patients with active sarcoidosis compared with those in control subjects. CONCLUSION: These results demonstrated that the level of sHLAs in serum is a useful index of disease activity of sarcoidosis, partly reflecting production by PBMCs and BALF cells.     

Clinical significance of serum KL-6 in pulmonary sarcoidosis]

We investigated whether the level of serum KL-6 could be an activity marker for pulmonary sarcoidosis. In 33 patients with pulmonary sarcoidosis, the relationships between serum KL-6 levels and diagnostic imaging, serum angiotensin-converting enzyme (ACE) levels, serum lysozyme levels, steroid therapy, and prognosis were evaluated. There were no significant differences in the level of serum KL-6 when the patients were divided on the basis of radiographic findings, but the level of serum KL-6 was markedly elevated in some patients with stage-II pulmonary sarcoidosis. There was a significant correlation between serum KL-6 levels and the following two parameters: serum ACE and lysozyme levels. Among patients with a high initial level of serum KL-6, pulmonary sarcoidosis tended to become exacerbated within one year. Steroid therapy significantly decreased the level of serum KL-6, suggesting that the level of serum KL-6 could be an activity indicator for pulmonary sarcoidosis. Immunohistochemical staining by anti-KL-6 antibody revealed that KL-6 was localized in proliferating type-II alveolar epithelial cells.     

Antibody activity to Propionibacterium acnes in bronchoalveolar lavage fluid in sarcoidosis

While increased levels of circulating antibody to various microorganisms have been reported in sarcoidosis patients, the pathogenesis of the disease is still unknown. In this report, the levels of antibody activities against Propionibacterium acnes (P. acnes) were measured in bronchoalveolar lavage fluid (BALF) in patients with sarcoidosis, using an enzyme-linked immunosorbent assay method. Each immunoglobulin class of antibody activity to P. acnes was corrected by albumin concentrations in BALF. The levels of whole immunoglobulin antibody activities to P. acnes in BALF were as follows: 412.3 +/- 443.9 O.D./albumin 1 mg (M +/- SD) in 31 untreated sarcoidosis patients, 556.6 +/- 341.8 in 10 sarcoidosis patients treated with prednisolone, and 231.5 +/- 156.8 in 16 control individuals. The levels of antibody activities were significantly elevated in untreated patients (p less than 0.05) and in treated patients (p less than 0.02) compared to those of controls. However, considering the treated vs. untreated patients, there was no significant difference in levels. The serum levels of whole immunoglobulin antibody activities were 0.484 +2- 0.191 O.D. in 38 untreated patients, 0.410 +/- 0.166 in 13 treated patients and 0.571 +/- 0.254 in 52 controls. The levels of antibody activity were significantly lower in treated patients than in the controls (p less than 0.05). However, there was no significant difference between the untreated patients and controls. To assess the site of antibody production, the secretion ratio was calculated by dividing the levels in BALF to those in serum. For this purpose, each serum level of antibody activity was also corrected by serum albumin concentration as with BALF.(ABSTRACT TRUNCATED AT 250 WORDS)