Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

IgE and IgG antibodies against Af were measured by ELISA in sera from 15 patients with ABPA and compared with antibody levels in six patients with extrinsic asthma and immediate-type skin reactivity to Af (prick skin test 3+ or 4+) but no other evidence for ABPA. The assay used polyvinyl microhemagglutination plates as a solid phase to adsorb antigens of Af and a double-antibody system of class-specific anti-human globulin and alkaline phosphatase-conjugated goat anti-rabbit globulin. IgE-Af and IgG-Af were significantly greater among patients with ABPA than in patients with asthma, suggesting that this assay may be used as an important diagnostic aid. The advantages of ELISA over radioimmunoassay are discussed in regard to early detection of an indolent pulmonary disease.
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PMID:Application of enzyme-linked immunosorbent assay (ELISA) in diagnosis of allergic bronchopulmonary aspergillosis. 703 11

Micropreparative two-dimensional (2-D) gel electrophoresis with immobilized pH gradients (4-8) in the first dimension (IPG-DALT) was optimized for the separation of salt-soluble wheat grain proteins, associated with bakers' asthma disease. The resolved polypeptides were electroblotted onto a polyvinylidene difluoride (PVDF) membrane and incubated with the pooled sera from four asthmatic bakers. Bound IgE was demonstrated by alkaline phosphatase conjugated anti-human IgE. Major IgE binding was detected in the 27 kDa, 37 kDa and, to a lesser extent, in the 14-18 kDa area of the 2-D immunoblots, respectively. Since the main purpose of our study was to determine the N-terminal amino acid sequences of the major wheat grain allergens, N-terminal sequencing was performed for six out of a total of eleven major allergens located in the 27 kDa area, for one out of two 37 kDa allergens, and for two out of four 14-18 kDa allergens. Our results revealed that two of the 27 kDa polypeptides are clearly related to several Acyl-CoA oxidase variants of barley and rice, whereas no significant homologies were found for the remaining four 27 kDa allergens analyzed. The N-terminus of the 37 kDa allergen appeared to be blocked so that no sequence information was obtained, while the two 14-18 kDa allergens analyzed were identified as members of the wheat alpha-amylase-inhibitor family.
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PMID:Sequence analysis of wheat grain allergens separated by two-dimensional electrophoresis with immobilized pH gradients. 749 55

The Kallestad technique for measurement of total IgE is based on a "sandwich" technique that uses a solid phase (wells in a sensitised microplate) as a separator. The samples that contain the IgE and monoclonal anti-IgE antibodies (AcM anti-IgE) are incubated in the wells sensitised with goat anti-mouse IgE. The AcM anti-IgE binds the IgG and the complex is then bound to the wells by the anti-mouse IgG. After washing, anti IgE marked with alkaline phosphatase is added and this binds to the previously linked IgE. The intensity of colouration is directly proportional to the concentration of IgE in the sample and is measured by spectrophotometry. The Kallestad technique has very great sensitivity (> 1 KUI/L) for a serum sample of 20 microliter. The measurement is quick, with 2 h incubation. There was good intra-assay reproducibility for values 2 to 1100 KUI/L with CV of 7.6. This good reproducibility was also found in inter-assays for values between 2 and 10 KUI/L. The Kallestad technique was evaluated firstly against the CAP RIA System of Pharmacia and secondly, for values against the kit RIA Ultra. For a population labelled "all comers" with values between 2 to 1100 KUI/L the statistical analysis showed no significant difference between the two techniques. The statistical analysis of the study of low values, made with the RIA Ultra kit showed that there was a significant difference between the two techniques. Exchange of standards between the two methods showed a certain over-evaluation of the values of Pharmacia whilst measurements with the Kallestad standards showed a relative under-evaluation. It should be noted that the comparison has been made between a RIA (Pharmacia) technique and one of EIA (Kallestad).
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PMID:[Total serum IgE levels. Comparative study of the Kallestad and Pharmacia techniques]. 770 27

Resort therapy-induced immunological changes were studied in 67 children from the unfavourable environmental areas, out of them 38 children (an experimental group) on Vetoron and 29 untreated children (a control group). The examinees were found to have secondary (acquired) immunodeficiency appeared as lower levels of immunoglobulin (Ig) A and higher levels of IgM, IgG and IgE. The health resort therapy improved the body's overall responsiveness--elevated the levels of IgA and IgM-reduced these of IgE, enhanced the activity of alkaline phosphatase and increased monocyte counts. The use Vetoron enhanced the efficiency of spa therapy in the examined children versus the matched ones, significantly elevated the relative levels of T lymphocytes and IgG and increased the activity of phagocytes.
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PMID:[The effect of water-soluble beta-carotene (Vetoron) on the immune status of children from ecologically unfavorable regions]. 770 13

Bronchial epithelial cells are activated in asthma but the mechanisms underlying this activation are poorly understood. We tested the possibility that bronchial epithelial cells recovered by brushing from 15 asthmatic and 11 control subjects may be activated by an IgE-dependent mechanism. The expression of the low-affinity IgE receptor (CD23) was studied by immunocytochemistry using the alkaline phosphatase anti-alkaline phosphatase technique and immunofluorescence using confocal microscopy. Four of eight allergic asthmatic patients and none of the seven non-allergic asthmatic or control subjects had a positive expression of CD23. The functional activity of CD23 was examined in the cells recovered from these subjects by stimulating them with IgE/anti-IgE. 15-HETE was not released but endothelin was released in the three or four asthmatic patients who had a positive expression of CD23. None of the other subjects released any endothelin. This study suggests that bronchial epithelial cells of asthmatic patients may be directly activated by an IgE-mediated mechanism.
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PMID:Low-affinity receptor for IgE on human bronchial epithelial cells in asthma. 783 11

Alveolar macrophages (AM) play a regulatory role in asthma. AM from asthmatics are activated, release increased amounts of cytokines, and express higher levels of the low affinity receptor for IgE (Fc epsilon RIIb/CD23b) and receptors for adhesion molecules. The bronchial microenvironment may modulate the phenotypic and functional characteristics of AM. On AM from normal subjects, the effects of histamine were studied on the expression of adhesion molecules (LFA-1, ICAM-1) and CD23b as well as on the release of fibronectin. The expression of LFA-1, ICAM-1, and CD23b was examined by immunocytochemistry using the alkaline phosphatase-anti-alkaline phosphatase technique. The expression of CD23b mRNA was studied by in situ hybridization. The release of fibronectin was measured by enzyme immunoassay. We found that histamine induced in a dose- and time-dependent fashion a significant increase of AM expressing the three membrane markers and a significant increase in the release of fibronectin. The maximum effect of histamine was observed after an incubation of 12 to 24 h and a dose of 1 microM. The histamine effects were specific, since they were significantly inhibited by an H1-blocker, pyrilamine, used at a concentration of 10 microM. The effect of an H2-blocker (ranitidine, concentration of 10 microM) was inconstant. Cycloheximide blocked the histamine effects, suggesting that it requires protein synthesis for its action. This study provides an in vitro model of cellular interaction between mast cells and AM, which might be relevant in the airway inflammation in asthma.
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PMID:Phenotypic and functional modulation of normal human alveolar macrophages by histamine. 791 13

A method for quantification of serum total IgE concentration in dogs by use of an ELISA containing monoclonal mouse anti-canine IgE was developed. Microtitration plates were coated with monoclonal mouse anti-canine IgE. Test sera and reference serum dilutions were added, followed by biotinylated monoclonal mouse anti-canine IgE. Avidin-alkaline phosphatase conjugate was added, and color development was measured spectrophotometrically, using a microtitration plate reader. Quantitative results were obtained by assigning to a reference serum a value of 100 IgE units/ml. Absorbance values of unknown samples were converted into IgE units by comparison with a standard curve generated by measurement of reference serum dilutions. Intra- and interassay coefficients of variation were 5 and 7%, respectively, and assay sensitivity was 1 U/ml. The assay was used to establish a normal range for total IgE concentrations in 30 healthy dogs. Total IgE concentration in healthy dogs followed a skewed distribution and ranged from < 1 to 91.2 U/ml, with a geometric mean value of 7.1 U/ml. The IgE concentration was remarkably stable in serum samples subjected to 25 freeze/thaw cycles or incubation at approximately 25 C (room temperature) for up to 10 days. Comparison of total IgE concentrations in 23 serum samples assayed by use of double-overlay radial immunodiffusion and ELISA yielded correlation coefficient of 0.94. Comparison of the reference serum standard curve with serial dilutions of a purified IgE solution of known concentration yielded a range of values for the IgE unit of 0.7 to 2.0 micrograms.
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PMID:Quantification of serum total IgE concentration in dogs by use of an enzyme-linked immunosorbent assay containing monoclonal murine anti-canine IgE. 797 32

This study was designed to analyse IgE and its related phenomena in bullous pemphigoid (BP). We analysed 17 BP sera by indirect immunofluorescence (IIF) and immunoblotting (IB) using a monoclonal antibody to IgE. In addition, inflammatory cells in lesional skin from 11 patients with BP were analysed by the alkaline phosphatase-anti-alkaline phosphatase (APAAP) technique using monoclonal antibodies to IgE and Fc epsilon RII/CD23. IgE class anti-basement membrane zone (BMZ) autoantibody was detected in nine of 17 sera (52.9%) by IIF. IgG class anti-BMZ antibody could block the BMZ-binding reactivity of IgE class antibody. Titres of IgE class autoantibody in the sera ranged from 1:40 to 1:320, and statistically correlated with serum IgE levels. Two of 11 sera contained an IgE class autoantibody which recognized a 230-kDa BP antigen by IB. By radio-allergosorbent test (RAST), IgE-specific antibodies to an extended series of common inhalant and food allergens were detectable in six sera with high concentrations of total IgE (over 3,300 IU/ml). IgE-bearing and Fc epsilon RII-expressing cells were demonstrated in the upper dermis and along the BMZ in seven of 11 biopsy specimens by the APAAP technique. The distribution and number of IgE-bearing cells in the lesions were similar to those of the Fc epsilon RII-expressing cells. These results suggest that both IgE-mediated immune responses and autoimmunity characterize BP as distinctive features.
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PMID:IgE and its related phenomena in bullous pemphigoid. 849 49

Fifteen sural nerve biopsies of vasculitic neuropathies have been compared with 11 cases of different non-vasculitic neuropathies and normal nerves from brain-dead organ donors. The APAAP (alkaline phosphatase monoclonal anti-alkaline phosphatase) immunostaining method was applied to cryostat sections from unfixed snap-frozen tissue samples. Immunoglobulins IgG, IgM, IgA, complement factors and light chains were reactive in biopsies of normal nerves as well as of vasculitic and nonvasculitic neuropathies. A strong reaction against IgE in the epineurial vessel walls was only seen in cases of Churg-Strauss-vasculitis. Antibodies against MHC class II (HLA DR) were positive in most of vasculitic infiltrates. Vascular endothelial cells were positive with anti MHC class I in all biopsies. A typical finding in all vasculitic neuropathies was the infiltration of epineurial vessels with CD4 positive and, to a lesser extent, CD8 positive lymphocytes. CD22 positive lymphocytes (B cells) have only been seen in about one third of vasculitic neuropathies. CD16 positive cells (NK-cells or neutrophils) could be demonstrated only in two biopsies. CD68 positive cells (macrophages) are frequently seen in most cases of neuropathy regardless of their etiology. The results support the concept of a primary T-cell mediated process against epineurial vessels as the most important mechanism in the pathogenesis of vasculitic neuropathies. In some cases with small epineurial infiltrates the vasculitic process can only be recognized with antibodies against CD4 or CD8. Therefore, the immunohistochemical evaluation of sural nerve biopsies may be helpful for identifying cases with microvasculitis.
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PMID:Immunohistochemical findings in vasculitic neuropathies. 850 63

For protein quantitation in gels or blotting membranes, chemiluminescence (CL) offers the advantages of a substantial improvement of detection limits. Easy-to-use CL-chemicals and specific probes such as antibodies conjugated to enzymes, e.g. alkaline phosphatase (AP) may be used in combination with a newly developed luminometer. CL was found to have low detection limits and a linear relation between relative light units (RLU) and the concentration of the antibody enzyme complex present over a wide concentration range. Measurements of the immunoglobulin IgE in dot blots and in blots after sodium dodecyl sulfate (SDS) electrophoresis under nonreducing conditions in agarose gels are described.
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PMID:Sensitive quantification of proteins by electrophoresis in gels by use of chemiluminescence. 852 3


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