Enzyme immunoassay plates should be shaken before absorbance is measured.
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Immunoenzyme Techniques↗
Biomedical subjects
S Husby
Publications and source records attributed to S Husby.
Circulating immune complexes in dermatitis herpetiformis and coeliac disease.
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Antigen-Antibody Complex↗
Immune complex formation analysed by high-performance size exclusion chromatography (HPLC-SEC) using either 125I-labelled antigen or enzyme-linked immunosorbent assay (ELISA) for detection.
A physical/immunochemical method has been developed for the analysis of soluble immune complexes. Human serum with high amount of antibody against bovine serum albumin (BSA) was incubated at 37 degrees with a wide range of 125I-labelled BSA and separated according to size by high-performance size exclusion chromatography (HPLC-SEC) on a TSK G 6000 PW column. Fractions were collected onto microplates and analysed either by gamma counting or by an enzyme-linked immunosorbent assay (ELISA) technique detecting anti-BSA antibody. The size of the immune complexes were the same when analysed by the two methods. The heaviest immune complexes were seen in moderate antibody excess where the majority of the complexes were eluted in a broad peak around Mr of 7 X 10(6). In large antibody excess the immune complexes eluted at around 3 X 10(6). Around equilibrium two peaks were seen corresponding to 3 X 10(6) and 5 X 10(5). In antigen excess the peak at 3 X 10(6) is diminished and free antigen appears. No changes in the distribution of the complexes were observed when the serum was made 20 mM in EDTA before incubation with BSA.
Adult↗
Chronic Pseudomonas aeruginosa lung infection in cystic fibrosis. A longitudinal study of immune complex activity and inflammatory response in sputum sol-phase of cystic fibrosis patients with chronic Pseudomonas aeruginosa lung infections: influence of local steroid treatment.
A double blind controlled trial of Becotide (beclomethasone diproprionate) inhalations was carried out for treating cystic fibrosis patients with chronic P. aeruginosa lung infection to determine its efficacy and safety. The aim of the treatment was to diminish the inflammatory response in the lungs of these patients, a response which is initiated by an allergic type III reaction. Pulmonary inflammation was evaluated by measurements of proteolytic activity, albumin concentration and immune complex activity in the sputum sol-phase before, during, and after the 16 weeks the trial lasted. 26 cystic fibrosis patients participated (13 received Becotide and 13 placebo) and the results showed that local steroids have no effect on the inflammatory response in the lungs of cystic fibrosis patients with chronic P. aeruginosa lung infection. No adverse effects were demonstrated. There was, however, a significant increase in the inflammatory parameters for all 26 cystic fibrosis patients when the trial period was over, compared to the insidious pulmonary destruction which takes place in the lungs of these patients, and it corresponded to a significant decrease (p less than 0.05) in forced vital capacity which took place at the same time. Therefore, chronic P. aeruginosa lung infection in these patients should be treated as efficiently as possible.
Adolescent↗
Molecular distribution of preformed thyroglobulin immune complexes.
Thyroglobulin anti-thyroglobulin immune complexes with human antibodies from four patient sera were performed in a wide range of antigen: antibody ratios and run in 5%-40% sucrose gradients. The fractionation showed simple distributions of thyroglobulin molecules in immune complexes, especially in far antigen and far antibody excess. A computer simulation analysis which utilized the experimental presumptions and assumed an association constant in the order of 10(9), showed a large degree of similarity to the experimental results. Thus, a thermodynamically simple explanation of immune complex formation did not contradict the experimental thyroglobulin anti-thyroglobulin complex formation.
Antigen-Antibody Complex↗
Hyponatraemia due to a thiazide diuretic. A case report.
The time course of profound hyponatraemia has been followed in a patient who developed this condition after 6 week's treatment with a thiazide diuretic, bendroflumethiazide. The case is related to the syndrome of inappropriate secretion of antidiuretic hormone.
Bendroflumethiazide↗
In vitro characterization of synthesized thyroglobulin immune complexes (IC). A model for studies of IC containing an autoantigen.
Human thyroglobulin (Tg) immune complexes (TgIC) were preformed at different ratios from purified Tg, and Tg antibodies (TgAb) obtained from six patients with autoimmune thyroid diseases. TgIC were characterized by precipitation with polyethylene glycol, and sedimentation in sucrose gradient. TgIC from one patient were further characterized by concanavalin A (con A), and binding to the complement factor Clq and rheumatoid factor (RF). Tg and TgIC had almost identical binding to con A, no binding to Clq, but could be partially separated by ultracentrifugation and polyethylene glycol. The highest degree of separation was obtained by a RF-coated plastic tube radiometric assay, using 125-I-rabbit TgAb as indicator. Tg showed no binding to RF, and a dose-response curve of TgIC in serum could be established. There was a dependence of the Tg-TgAb ratio, TgIC at equilibrium and in antibody excess being detected most efficiently. The method may serve as an aid in the evaluation of the fate of Tg, TgAb and TgIC following thyroid damage (surgery, radioiodine) and may be extended as a model system in the investigation of immune complexes in connection with autoimmune disorders.
Antigen-Antibody Complex↗
Methodological approaches to antigen identification in soluble immune complexes; A model study.
The usefulness of different techniques for identification of immune complex-(IC) bound antigen(s) was investigated in two Ab-Ag model systems. The following approaches were used: 1) determination of IC-activity before and after removal of IC by adsorption to antibody-specific solid phase systems. 2) analysis of PEG-insoluble IC by reference line-rocket immunoelectrophoresis and 3) reprecipitation of PEG-insoluble IC in the presence of 125I-labelled antibody to suspected antigens and subsequent analysis of the distribution of the radioactivity. This latter approach, the use of a double PEG precipitation immunoradiometric assay, was found most useful and simple for identification of antigen(s) contained in soluble IC. The technique is most sensitive in demonstrating IC-bound antigen(s) when the IC are formed in slight to moderate antigen excess.
Adsorption↗
[Determination of immune complexes in serum with the complement consumption test].
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Adolescent↗
Soluble immune complexes in cerebrospinal fluid of patients with multiple sclerosis and other neurological diseases.
The occurrence of soluble immune complexes (IC) in the cerebrospinal fluid (CSF) of 14 multiple sclerosis (MS) patients, four acute polyradiculoneuritis patients, 30 patients with other neurological diseases (OND) and 30 patients with disc prolapse (DP) was examined by a solid phase C1q-protein A binding assay (C1q-PABA) and a complement consumption test. IC-positive reactions were observed only in the C1q-PABA. The binding indices determined by the C1q-PABA differed significantly (P < 0.01) when the MS or the OND patient groups were compared to the DP group. No significant (P < 0.1) difference was observed between the indices in the MS and OND groups. Binding indices in C1q-PABA showed no correlation either to IgG concentration, total protein concentration or cell counts in CSF of MS patients. Three of the four polyradiculoneuritis patients were strongly IC-positive while the fourth patient was negative. Filtration and PEG-precipitation data indicated that a major part of the IgG-containing IC in CSF detected by C1q-PABA was of macromolecular nature.
Adolescent↗
Two pregnancy-associated serum proteins with pregnancy-specific beta1-glycoprotein determinants.
A protein showing strong antigenic cross-reactivity with the "pregnancy-specific" beta1-glycoprotein (PSbetaG) was demonstrated in serum of pregnant women during the second and third trimesters, but not in sera from normal men and nonpregnant women. The described protein had alpha2-electrophoretic mobility and its molecular weight was estimated to be around 200,000 as compared to 80,000 for PSbetaG. Crossed immunoelectrophoretic analysis, using antiserum to PSbetaG, indicated the presence of additional antigenic determinants in PSbetaG not demonstrable in the cross-reacting alpha2-protein. The PSbetaG/alpha2-protein ratio was rather constant in consecutive serum samples from one woman during pregnancy, but showed a marked variation when sera from different women were compared. Results of rocket immunoelectrophoresis gave a rough estimate of the ratio between these two proteins in a sample of pregnancy serum. The reported findings point to the need for development of specific methods for quantitation of the two proteins and the necessity to re-evaluate earlier quantitative PSbetaG data.
Blood Protein Electrophoresis↗
Effect of a single graded dose of terbutaline tablets in patients with chronic bronchitis and bronchoconstriction. A double-blind, placebo, cross-over study.
Terbutaline, a selective beta2-adrenergic receptor stimulator was given to 10 patients with chronic bronchitis. The effects of the drug were tested by measurements of forced expiratory volume in 1 sec (FEV1), peak expiratory flow rate (PEFR), heart rate, blood pressure and blood gas analysis. The measurements were performed before and 1, 2, 4, 5, and 6 h after oral administration of placebo, 2.5 mg, or 5.0 mg terbutaline. Terbutaline caused a significant dose-related increase in FEV1 and PEFR as compared with placebo. The maximal effects were found at the 4-h measurement and were still present at the measurements performed at 6 h. Heart rate, blood pressure and arterial oxygen tension (PaO2) were not significantly affected. Four patients experienced side effects as tremor and/or heart palpitations. It is concluded that orally administered terbutaline may be an important therapeutical agent in the treatment of chronic bronchitis.
Aged↗
Genetic influence on the serum levels of naturally occurring human IgG antibodies to dietary antigens. Quantitative assessment from a twin study.
Serum IgG antibodies to ovalbumin (OA) and beta-lactoglobulin (BLG) were quantified by ELISA techniques in 22 monozygotic (MZ) and 24 dizygotic (DZ) healthy twin pairs. Antibody levels were comparable in the MZ and DZ groups both for anti-OA and anti-BLG antibodies. The genetic variance (GWT) was 0.167 for log IgG anti-OA antibodies, and 0.173 for log IgG anti-BLG antibodies, with heritability estimates of 0.44 and 0.37, respectively. No indication was observed of genotype-environmental interaction or differential environmental covariance for the log antibody levels in the MZ and DZ twins. The anti-OA and anti-BLG antibody levels in the same individual correlated only to a low degree. The levels of naturally occurring serum IgG antibodies are significantly influenced by genetic factors.
Adolescent↗