儿童变应性支气管肺曲霉菌病影像学研究进展

变应性支气管肺曲霉菌病（ABPA）为免疫介导肺部疾病。儿童ABPA发病率较低,且临床特征相比成人较不典型,导致诊断难度较大。高分辨率CT是ABPA的首选影像学检查方法;MRI近年也逐渐用于诊断ABPA。本文基于ABPA发病机制及临床表现对儿童ABPA肺部影像学研究新进展进行综述。     

Mucus plugs in peripheral bronchi: An unusual initial manifestation of allergic bronchopulmonary aspergillosis

Although allergic bronchopulmonary aspergillosis can be associated with mucus plugs in the central bronchi, this association in the peripheral bronchi remains unclear. A 78‐year‐old woman presented with mucus plugs in both the peripheral and the central bronchi in the right lung, which evolved into consolidation with high‐attenuation mucus after one month.     

Dexamethasone and cyclosporin A modulation of cytokine expression and specific antibody synthesis in an allergic bronchopulmonary aspergillosis murine model

We previously demonstrated that, in C57B1/6 mice, cyclosporin A enhanced and dexamethasone inhibited the Aspergillus fumigatus -induced pulmonary eosinophilia and total IgE levels. To evaluate whether these effects were related to the modulation of T-lymphocyte recruitment and activation and cytokine expression, we performed immunohistochemical staining for T-cell surface marker CD3 and CD4, cell activation marker CD25, and cytokines granulocyte–macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL-4) and interleukin-5 (IL-5) on lung tissue sections from mice exposed to Aspergillus fumigatus and treated or not with dexamethasone or cyclosporin A. Dexamethasone significantly inhibited Aspergillus fumigatus -induced increased number of activated T cells and cytokine-expressing cells in parallel with a decrease in pulmonary eosinophils. In contrast, cyclosporin A did not decrease these immunological events but enhanced the lung eosinophil recruitment. Moreover, dexamethasone prevented the production of immunoglobulins against 76 and 36 kD antigen proteins and cyclosporin A against 76 and 18 kD antigen proteins. These results indicate that dexamethasone down-regulates and cyclosporin A up-regulates lung eosinophil recruitment and total IgE production, probably via the modulation of T-lymphocyte activation and GM-CSF, IL-4 and IL-5 expression. Both drugs inhibit Aspergillus fumigatus -specific antibody synthesis, but their suppressive actions are selective to different antigenic components.     

用侵袭性曲霉病患者血清筛选特异性反应的烟曲霉蛋白质

目的 用对烟曲霉蛋白质反应阳性的侵袭性曲霉病(IA)患者血清筛选免疫反应性最强的烟曲霉蛋白质用于免疫蛋白质组分析.方法 TCA/丙酮沉淀法制备烟曲霉分泌蛋白及菌体蛋白质作为包被抗原,用ELISA法筛选确诊IA患者血清,反应阳性的血清进一步与烟曲霉分泌蛋白及菌体蛋白质进行免疫印迹,筛选免疫反应最强的蛋白质.结果 11例确...     

Serologic and cellular assays to monitor therapy with murine monoclonal antibodies

The emergence of new therapeutic modalities frequently requires development of relevant laboratory assays. This is especially true for the clinical uses of newly developed reagents such as murine monoclonal antibodies. We have described immunofluorescence, immunohistochemical, and enzyme-linked immunosorbent assays (ELISA) for monitoring anticancer therapy with monoclonal antibodies. Immunofluorescence assays performed on single cell suspensions and immunohistochemical assays performed on freshly frozen biopsy tissue have confirmed tissue reactivity of the monoclonal antibody prior to infusion and have been used to measure in vivo antibody binding to target cells as well as antigenic modulation during therapy. An ELISA has been used to measure the serum concentrations of monoclonal antibodies; another ELISA has been used to detect the presence of human antibodies reactive with murine monoclonal antibody. These reproducible assays can be performed with commercially available reagents and provide an important data base for making clinical decisions concerning therapy with monoclonal antibodies.     

Analysis of HIV seropositive thalassemic children for antibodies specific to Aspergillus fumigatus by luminescent immunoassay

The applicability of luminescent immunoassay (LIA) in serodiagnosis of fungal infections in multitransfused (MT) thalassemic children seropositive for human immunodeficiency virus (HIV) was investigated. Thirty-one sera samples from HIV infected pediatric patients with thalassemia receiving repeated blood transfusions were analysed for the presence of antibodies specific to Aspergillus funigatus by LIA. The LIA was standardized using well defined antigens of A. Fumigatus. Ten out of 31 (32.2%) of the MT-HIV positive patients were found to have anti-Aspergillus antibodies in their sera by LIA. The ELISA could detect A. Fumigatus specific antibodies in 25.8% (8 out of 31) of the patients. Thus, 20% more number of patients turned to be positive for aspergillosis by LIA as compared to ELISA. The difference was found to be statistically significant (p < 0.005). Of the MT-HIV negative patients only 1 out of 33 (3%) showed A. Fumigatus specific antibodies by LIA and ELISA both. In age and sex matched control group (n = 25) none of the patients was found to be positive for antibodies to A. Fumigatus. LIA was found to have better discriminatory value indicating, thereby, its utility in diagnosis of aspergillosis in compromised patients. J. Clin. Lab. Anal. 11:343–345, 1997. © 1997 Wiley-Liss, Inc.     

抗-TP 抗体临床检测结果分析及感染趋势评估

目的：了解广西钦州地区梅毒螺旋体（TP）临床感染状况,并分析其感染趋势。方法采用酶联免疫吸附试验（ELISA）对该院2009～2012年住院及门诊患者血液标本进行抗-TP 抗体检测。结果4年间共检测19362份标本,抗-TP 抗体阳性率为6．13％,性病泌尿科的阳性率高达29．42％,且逐年增长,其差异有统计学意义（χ2＝9．738,P ＜0．05）。女性的抗-TP 抗体阳性率（7．03％）高于男性（5．47％）,20～40岁组抗-TP 抗体阳性率最高。结论近年来钦州地区 TP 感染率逐年递增,应在预防宣传上加大力度,防止梅毒的进一步扩散。     

Screening and characterization of specific anti-lipopolysaccharide antibodies in Belgian blood donors by enzyme-linked immunosorbent assays

The goal of this project was to find and collect high concentrations of endotoxin-specific antibodies for therapeutic IgG- or IgM-enriched preparations. Various enzyme-linked immunosorbent assays (ELISAs) were developed to perform longitudinal studies of the serological response to a large panel of smooth and rough purified lipopolysaccharide (LPS) extracts in a population of healthy blood donors. To accomplish this, 1612 human serum samples from volunteer blood donors collected by seven different blood banks in Belgium were screened and specific IgM and IgG activities were measured. Approximately 17% of the donors had anti-LPS concentrations higher than 40 mg L⁻¹. Of these, 10.9% had anti-smooth LPS antibodies, 3.7% had anti-rough LPS antibodies and 2.8% were found to be positive towards both types of LPS. The mean anti-LPS antibody concentration was 8 mg L⁻¹ for rough LPS and 14 mg L⁻¹ for smooth LPS. Age- and sex-related distributions of the activities indicated that the greatest prevalence of high anti-LPS concentration was in women aged 40–49 years and in men older than 60 years. Differential absorption experiments showed that the pooled serum of selected blood donors contained a mixture of specific and cross-reacting antibodies. We detected predominantly anti-LPS activities due to the IgG₁ and IgG₂ subclasses. The range of specificities to different LPS was increased by the pooling of selected sera. It was concluded that pools of naturally occurring specific anti-LPS immunoglobulin antibodies may be obtained in Belgium by screening blood donors using ELISAs that we have developed.     

A new insulin immunoassay specific for the rapid-acting insulin analog, insulin aspart, suitable for bioavailability, bioequivalence, and pharmacokinetic studies

Objectives: To validate a specific enzyme-linked immunosorbent assay for the rapid-acting human insulin analogue, insulin aspart, in human serum, human plasma, and porcine plasma.

Design and methods: For the enzyme-linked immunosorbent assay, two murine monoclonal antibodies were developed that bind to two different epitopes on the insulin aspart molecule. Key parameters for validation were imprecision, accuracy, matrix effects, dilution-linearity, and cross-reactivity.

Results: No cross-reactivity was found with human and porcine insulin, human proinsulin, or human C-peptide. The assay is sensitive (limit of QUANTIFICATION = 11.5 pmol/L), accurate (95–107% recovery with human serum, human plasma, and porcine plasma in the range 16–800 pmol/L), and has a 14.7% total imprecision within the entire analytical range. Dilution of samples gave linear results with human serum as the diluent.

Conclusions: The insulin aspart-specific enzyme-linked immunosorbent assay described in this study is well suited to study the bioavailability, bioequivalence, and pharmacokinetics of this insulin analogue.     

抗核小体抗体对系统性红斑狼疮诊断价值的探讨

目的：探讨抗核小体抗体对系统性红斑狼疮的诊断和判断疾病活动度的价值。方法：采用酶联免疫吸附试验（ELISA）检测45例系统性红斑狼疮、25例类风湿关节炎、16例原发性干燥综合征、8例多发性肌炎／皮肌炎、5例混合性结缔组织病、18例骨性关节炎和30例健康对照组血清中的抗核小体抗体，将抗核小体抗体与疾病活动度（以SLEDAI评分）、抗双链DNA抗体、抗Sm抗体等指标进行比较。结果：抗核小体抗体在45例系统性红斑狼疮患者34例（75．5％）阳性，其敏感度和特异度分别为75．5％，93，1％；抗核小体抗体与SLEDAI评分、抗双链DNA抗体、肾损害有相关性。结论：抗核小体抗体在系统性红斑狼疮中敏感性较高，是诊断系统性红斑狼疮和了解狼疮活动度的良好指标。     

Microsatellite typing of Aspergillus fumigatus isolates recovered from deep organ samples of patients with invasive aspergillosis

Microsatellite typing was used to analyze 41 Aspergillus fumigatus isolates from 9 patients with proven invasive aspergillosis hospitalized in 2 different centers. No strains were shared between patients. For 8 of 9 patients, a single genotype was found for the isolates recovered from all anatomic sites involved.     

Uptake and efflux kinetics,and intracellular activity of voriconazole against Aspergillus fumigatus in human pulmonary epithelial cells: a new application for the prophylaxis and early treatment of invasive pulmonary aspergillosis

Invasive pulmonary aspergillosis (IPA), most caused by Aspergillus fumigatus, is a serious life‐threatening infection in immunocompromised patients. Voriconazole is used to prevent and treat IPA. However, little is known about the pharmacological characteristics of voriconazole in pulmonary epithelial cells, which are the target site for the prophylaxis and early treatment of IPA. The aim of the study was to evaluate the kinetics and activity of voriconazole against A. fumigatus in A549 cells. High‐performance liquid chromatography/tandem mass spectrometry and time‐kill method were used to study the cellular pharmacokinetic and pharmacodynamics of voriconazole. Voriconazole exerted a concentration‐dependent toxic effect on A549 cells and could penetrate into cells, reaching plateau concentrations of 1.14 ± 0.64, 3.72 ± 1.38 and 6.36 ± 0.95 ng/mg protein after A549 cells were exposed to voriconazole at extracellular concentrations of 2, 8 and 16 mg/L for 2 h, respectively. The efflux of voriconazole was rapid, with a half‐life of 10.2 min. Voriconazole can decrease the A. fumigatus conidia invade cells, and the number of viable A. fumigatus conidia in cells can be decreased 2.1‐ to 20.6‐fold when A549 cells were cultured in medium containing voriconazole. After 24‐h incubation, 75.6% and 80.5% of intracellular A. fumigatus were killed when extracellular voriconazole concentration was 8 and 16 mg/L, respectively. This study illustrated a new application for the prophylaxis and early treatment of IPA from the cellular pharmacokinetics and pharmacodynamics and emphasized the importance of monitoring concentrations of voriconazole in epithelial lining fluid in immunocompromised patients receiving voriconazole therapy.     

提高侵袭性烟曲霉感染痰标本病原学检出率的方法探讨

目的:探讨一种可提高临床痰标本的侵袭性烟曲霉感染检出率的培养基。方法:收集临床常规痰标本分离菌(包括白假丝酵母菌),分析其耐药性,并将其作为"干扰菌"与烟曲霉标准株的孢子以不同比例混合接种培养,模拟痰标本的混合感染,观察彼此的干扰情况。同时添加抗菌药物于MediumB培养基中,分析培养基对烟曲霉的生长选择性。结果:当铜绿假单胞菌浓度≥104CFU/mL时,即可完全抑制烟曲霉(103CFU/mL)的生长;金黄色葡萄球菌和大肠埃希菌则必须在>105 CFU/mL时,才能完全抑制103 CFU/mL的烟曲霉生长;当白假丝酵母菌与烟曲霉以100∶1混合接种时,其基本抑制了烟曲霉的生长。根据"干扰菌"的耐药特性,同时添加亚胺培南和万古霉素至培养基,结果显示其能有效抑制上述3种细菌的生长;添加氟康唑5μg/mL可抑制白假丝酵母菌的生长,而上述3种药物对烟曲霉的生长均无影响。结论:在所研究的真菌分离培养基中添加适宜的抗细菌及抗真菌药物,可有效抑制"干扰菌",提高烟曲霉的检出率。     

武汉地区变应性鼻炎患者常见变应原sIgE检测分析

目的:分析本地常见变应原分布特征,为合理防治变应性鼻炎提供依据。方法:利用间接酶联免疫冷光分析技术,对2018年7月至2019年9月间在武汉大学人民医院进行变应原特异性免疫球蛋白E（sIgE）筛查的1 190例变应性鼻炎患者进行血清学检测,其中,男性708例,女性482例。按年龄将受试患者分为0~6岁、7~17岁、18...     

Serologic diagnosis of allergic bronchopulmonary aspergillosis in patients with cystic fibrosis through the detection of immunoglobulin G to Aspergillus fumigatus

Allergic bronchopulmonary aspergillosis (ABPA) is seen in approximately 10% of patients with cystic fibrosis (CF) and can be difficult to diagnose. Consensus criteria require the presence of multiple elevated immunologic markers such as total immunoglobulin E (IgE), Aspergillus IgE and Aspergillus IgG, or precipitins for a robust diagnosis. There is some degree of standardization of total IgE and Aspergillus IgE levels, but there is no standardization in the measurement of IgG antibodies or precipitins to Aspergillus. The interpretation of results may, therefore, be confusing. Eighty-seven patients with CF were categorized as having ABPA or as controls, using the consensus criteria and an in-house enzyme immunoassay to measure IgG levels to Aspergillus. All sera from patients were then analyzed by commercial fluorescent immunoassay (FEIA) for the quantitative detection of anti-Aspergillus IgG. FEIA results were analyzed against the consensus conference minimum diagnostic criteria to ascertain a cutoff point, which could predict a diagnosis of ABPA in CF. Eighty patients with CF and with no or incomplete evidence of ABPA had a mean FEIA score of 51.1 mg/L, whereas 7 CF patients with ABPA had a mean FEIA score of 132.5 mg/L. Using receiver operator characteristic curve analysis of the ImmunoCAP (Phadia) IgG score on ABPA versus all other patients gave an area under the curve of 0.933 (estimated SE, 0.027). This analysis provisionally suggested that a score of 90 mg/L may be used as a cutoff point, which would give a sensitivity of 91% and specificity of 88.0% for the diagnosis of ABPA, though this requires further validation. This quantitative approach to Aspergillus IgG measurement in patients with CF along with the results of other tests will hopefully provide a more accurate approach to the diagnosis of ABPA.     

三种梅毒螺旋体抗体检测方法的比较分析

目的比较化学发光法（TP-CLIA）、酶联免疫吸附试验（TP-ELlSA）和梅毒螺旋体明胶颗粒凝聚试验（TPPA）对梅毒螺旋体特异性抗体（TP-Ab）检测的意义。方法分别用CLIA、ELISA和TPPA检测患者的1797份血清样本,收集CLIA／ELISA／TPPA／法检测均阳性但临床未明确诊断的标本及结果不一致标本以重组免疫印迹法（RIBA）最终确认。结果三种方法共筛选69例阳性及可疑血清标本。CIIA／EIISA／TPPA法均阳性标本63例,其中52例有临床明确诊断。另11例3种方法检测均阳性但未明确诊断标本和6例检测结果不一致标本用RIBA确证。CLIA法确认阳性66例,阳性率3．67;ELISA法确认阳性65例,阳性率3．62;TPPA法法确认阳性61例,阳性率3．39;CLIA、ELISA及TPPA法敏感性分别为98．51％、97．02％和91．05％;特异性均为99．88;诊断效率分别99．83％、99．78和99．66％。结论CLIA法和ELISA法敏感性均高于TPPA法,不管用何种方法检测对于临床诊断不符的标本均应慎重,必要时用RIBA法补充确认,以排除假阳性。     

Analysis of antibodies to Aspergillus fumigatus antigens by class-specific enzyme-linked immunosorbent assay in patients with pulmonary aspergillosis

Antibodies to two commercial extracts of Aspergillus fumigatus were determined by a class-specific enzyme-linked immunosorbent assay (ELISA) in 203 serum samples from 139 patients with various pulmonary diseases. In 22 patients with aspergilloma immunoglobulin (Ig)M, IgA, and especially IgG antibodies were found, whereas in 50 patients with allergic alveolitis IgG antibody was most frequent, IgM occurring rarely. One patient with allergic bronchopulmonary aspergillosis demonstrated IgG and IgA antibodies. Of 20 cases with bronchial asthma, 10% reacted against A. fumigatus in immunodiffusion as well as in ELISA. Of 46 cases with carcinoma, tuberculosis, and miscellaneous pulmonary diseases, 17% were positive by immunodiffusion and 26% demonstrated antibodies usually IgG, by ELISA. Of 100 healthy blood donors, none had Aspergillus antibodies of the IgG class, whereas 3% were positive in the IgM and 3% in the IgA assay. The ELISA proved to be sensitive and useful in the follow-up of patients with aspergilloma after operation.     

A simple procedure for the rapid immunofluorescence staining of human tumor antigens with human monoclonal antibodies

A novel technique employed in preparing slides used for immunofluorescence staining of human tumor antigens with human anti-cancer monoclonal antibodies is described. The anchorage of the cancer cells to the plastic substratum is used as a basis in eliminating cell fixation by physical and chemical procedures. In this method, the bottoms of the tissue culture flasks with the adherent A431 cell line, a carcinoma of the vulva, were cut into slides of 2 × 6 cm. This antigen-positive target cell was then reacted with the VLN3G2 monoclonal IgG antibody and with the fluorescein-labeled F(ab′)₂ of goat anti-human IgG. Most A431 cells demonstrated intense fluorescence staining while others exhibited diffuse and often discontinuous membrane staining with the antibodies. In summary, this simplistic technique can be applied for the rapid detection of antigens displayed by the adherent tumor cell lines.     

The monoclonal antibody-specific immobilization of erythrocyte antigens assay (MAIEA) in the investigation of human red-cell antigens and their associated membrane proteins

The monoclonal antibody-specific immobilization of erythrocyte antigens (MAIEA) technique is an immunoassay devised primarily for locating blood group antigens on specific red-cell membrane proteins. The assay involves the incubation of intact red cells with two antibodies, one human alloantibody, the other a nonhuman antibody, usually a rodent monoclonal antibody, but polyclonal antibodies of rabbit origin have been utilized. For a positive result, both antibodies must bind to the same membrane protein. The red cells are lysed, the membrane solubilized and the trimolecular complex of two antibodies and membrane protein is captured in a well coated with goat antirodent (or rabbit) immunoglobulin. The immobilized complex is then detected by the use of peroxidase-conjugated goat antihuman (or rodent) immunoglobulin.
Negative results, due to mutual blocking between the human and animal antibodies when their epitopes are close together on the same molecule, have permitted a degree of localization of epitopes on some proteins. This has been most effective in the mapping of Cromer blood group system antigens on the complement control protein domains of decay-accelerating factor (DAF, CD55), but has also proved informative in the clustering of antigens on the Lutheran and Kell glycoproteins.
MAIEA is an effective tool for the identification of antibodies to Knops-system antigens on complement receptor 1 (CR1, CD35) in immunohaematology reference laboratories. These antibodies are clinically unimportant, but must be identified before they can be ignored for transfusion purposes.