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Biomedical subjects

E M Salonen

Publications and source records attributed to E M Salonen.

70 records · Page 4Linked to original sources

Cerebrospinal fluid IgG bands and virus-specific IgG, IgM, and IgA antibodies in herpes simplex virus encephalitis.

To characterize the immune response of the central nervous system in herpes simplex virus (HSV) encephalitis, cerebrospinal fluid (CSF) specimens of 7 biopsy proven and 7 presumptive herpes simplex virus (HSV) encephalitis patients were studied, using sodium dodecyl sulfate polyacrylamide gel electrophoresis for the presence of CSF IgG bands, and solid-phase enzyme immunoassays for HSV-specific antibodies. IgG bands were detected in all CSF specimens of the patients, as early as day 6 and up to day 1088. A novel, unidentified, 120 000 dalton polypeptide was found in the CSF of most of the patients, in a total of 25/50 specimens, but not in the controls. This polypeptide was evident by day 6, its intensity fluctuated and it was present in specimens collected as late as day 855. HSV-specific antibodies, of either IgG, IgM, or IgA class, were not detected in the CSF during the first week of illness. IgG antibodies appeared later in all patients and persisted to the end of the follow-up of 3 years. The fact that CSF IgG bands were present in some patients before the appearance of HSV antibodies, and also persisted longer, suggests that the IgG response is not restricted to HSV-specific antigenic determinants.

Adult↗

Trial with human leucocyte interferon and vidarabine in herpes simplex virus encephalitis: diagnostic and therapeutic problems.

A combination therapy of human interferon, vidarabine, and dexamethasone was administered to six patients with proven (Patients I-III) or presumed (Patients IV-VI) herpes simplex virus encephalitis (HSVE). Interferon combined with dexamethasone was given to one patient with presumed HSVE (Patient VII). Leucopenia and elevated serum transaminase levels appeared in all patients and a diffuse bleeding in one of them. Patients II, III and IV died, 26, 43, and 209 days after the onset of encephalitis, respectively. Patients I, V, VI, and VII were left with moderate brain damage although their physical condition was good. HSV encephalitis presents diagnostic difficulties, complementary diagnostic methods are needed, and current therapeutic trials must be considered as preliminary.

Adult↗

C-reactive protein in acute viral infections.

A sensitive solid-phase enzyme immunoassay procedure was used to determine the concentrations of C-reactive protein (CPR) in the acute and convalescent phase sera of patients with verified rubella, herpes simplex, cytomegalo, influenza A or B, enterovirus, or mycoplasma infection. In all infection groups about 90% (80% for influenza) elevated CRP values were observed in the acute phase sera (mean values in the different groups 16-57 micrograms/ml), the highest values exceeding or approaching 100 micrograms/ml. The serum CRP values were highest in all groups before the specific serum antibodies were detectable and decreased approaching the upper limit or normal controls (2 microgram/ml) within 2 weeks. Notable individual variation in the CRP production was seen. We conclude tha serum CRP determination should not be used as a reliable criterion to distinguish bacterial and viral infections.

C-Reactive Protein↗

Local production of mumps IgG and IgM antibodies in the cerebrospinal fluid of meningitis patients.

Immunoglobulin G (IgG) and M (IgM) antibodies against mumps virus were measured by an enzyme-linked immunosorbent assay (ELISA) in the serum and cerebrospinal fluid (CSF) specimens of patients with mumps meningitis. The CSF IgG antibodies correlated well with the respective antibody titers in serum. On the contrary, in only about half of the patients a moderate correlation was found between the CSF and serum IgM antibody titers, while the other patients did not have detectable mumps IgM antibodies in CSF irrespective of intermediate to high titers in serum. Two different immunologic mechanisms may be involved in these two groups which, however, did not show any clinical differences. The lack of IgM antibodies in the CSF of many patients diminished the value of CSF in the laboratory diagnosis of mumps meningitis compared to use of serum specimens. Intrathecal synthesis of mumps IgG antibodies was demonstrated in 83% of the patients, and of IgM antibodies in at least 67% of those patients with detectable IgM antibodies in CSF. The ratio between mumps IgG and IgM antibodies was higher in CSF than in serum, suggesting that the synthesis of IgG antibodies in central nervous system was more efficient than that of IgM antibodies.

Antibodies, Viral↗

Rapid solid-phase enzyme immunoassay for antibodies to viruses and other microbes: effects of polyethylene glycol.

A rapid and sensitive solid-phase enzyme immunoassay (EIA) was developed for determination of serum antibodies, including addition of 4% (w/v) polyethylene glycol (PEG) to the diluent buffers. This modification accelerated the antigen-antibody reactions and made it possible to complete the assay within 2 h and to incubate at room temperature only. The enhancing effect of the polymer was particularly prominent in the second solid-phase immune reaction, the interaction between the antigen-bound antibody and the enzyme-labeled anti-immunoglobulin. The PEG-EIA procedure was successfully applied in the assay of antibodies to rubella, influenza A, herpes simplex and cytomegalo viruses, to Mycoplasma pneumoniae and Toxoplasma gondii.

Antibodies↗

Evaluation of solid-phase enzyme-immunoassay procedure in immunity surveys and diagnosis of rubella.

In the development of optimized micro-and macromethods for solid-phase rubella enzyme-immunoassay (EIA), the effect of various technical parameters was studied. Special emphasis was placed on elimination of defective polystyrene material, purity and quantity of immobilized antigen, use of reference sera in each microplate or macroprocedure, use of nonionic detergent in washing and diluent buffers, and use of a colourless enzyme substrate. A single serum dilution, analysed in duplicate, was sufficient provided the procedure was standardized. In quantitation of serum rubella antibodies, EIA gave values correlating well with those obtained with the single radial haemolysis test but not with low haemagglutination inhibition titres. The EIA procedure proved to be a useful and reliable serological tool in immunity surveys. When complemented by a rubella IgM test, EIA could also be used in diagnosis of recent rubella infections as a convenient primary serological test.

Adult↗

Rheumatoid factor in acute viral infections: interference with determination of IgM, IgG, and IgA antibodies in an enzyme immunoassay.

IgM rheumatoid factor (RF), an autoantibody to the Fc fragment of IgG, was determined by solid-phase enzyme immunoassay (EIA). RF levels were significantly higher in patients with rubella virus infection than in patients with infections due to influenza virus, cytomegalovirus, respiratory syncytial virus, parainfluenza virus, adenovirus, mumps virus, or herpes simplex virus. To evaluate the role of RF in EIA determinations of viral antibodies, IgM RF from IgM-IgG cryoglobulin or control IgM was added to patient sera before assay for viral antibodies. IgM RF inhibited virus-specific IgG and IgA reactions and gave nonspecific IgM reactions in EIA for antibodies to rubella and influenza viruses, but had little or no effect on antibodies to cytomegalovirus. The minimal effective amounts of RF were 100-500 ng/ml for inhibition of IgG, 300-1,000 ng/ml for IgA, and 25-500 ng/ml for IgM. The control IgM preparation gave no such effects. These studies reinforce the need to eliminate RF interference in solid-phase EIA.

Acute Disease↗

Rubella antibody determination from heparinised finger-tip blood by single radial haemolysis and enzyme immunoassay.

Rubella antibodies were determined by single radial haemolysis (SRH) and a micromodification of enzyme immunoassay (EIA) of samples of heparinised finger-tip blood or plasma collected into transportable vials, and the results were compared with antibody titres obtained from conventional samples of venous serum. The antibody titres of finger-tip specimens gave a high correlation with those of venous serum. For SRH only 5 microliters of heat-inactivated finger-tip plasma was needed, and for EIA only a single dilution, 1:100, in duplicate, of heparinised finger-tip plasma or while blood was sufficient. The minimal inconvenience in sample collection makes the finger-tip test particularly suitable for large-scale immunity screening when assessing the need for, or efficacy of, rubella vaccination.

Antibodies, Viral↗

Immobilization of viral and mycoplasma antigens and of immunoglobulins on polystyrene surface for immunoassays.

The immobilization of purified influenza virus, rubella virus, crude nuclear cytomegalovirus antigen and of mycoplasma on polystyrene tubes was studied using radio-iodinated preparations. The antigen activities on tube surfaces were determined using sequentially specific human antibodies and alkaline phosphatase-conjugated anti-human IgG in an enzyme-immunoassay (EIA) reaction. In addition, immobilization of radio-iodinated human IgG, IgM, and IgA, serving as model proteins, was studied using the respective anti-immunoglobulin conjugates in EIA directly. Pretreatment of the surface with albumin and glutaraldehyde inhibited the adsorption and antigenicity of IgG. Increase of temperature and thus of speed of adsorption did not affect the fraction of antigen eluted during the test procedure. Only with IgG and IgA was it necessary to saturate the polystyrene surface in order to achieve maximal reactivity in EIA. With other antigens, maximal reactivity in EIA was obtained with amounts of protein much lower than the maximal amount that could be adsorbed per tube. IgM was found to have an exceptionally high affinity to polystyrene.

Antigens, Bacterial↗

Porcine serum cobalophilin and transcobalamin. Identification, isolation and properties including electrofocusing patterns.

Pooled porcine serum was found to contain cobalophilin (also called transcobalamin I) and transcobalamin (also called transcobalamin II). The two proteins were harvested by batchwise absorption with vitamin B-12 covalently coupled to Sepharose, and then separated from each other either by gel filtration or using an immunoadsorbent. Both proteins were finally isolated as single proteins using a second vitamin B-12-Sepharose chromatography step. Cobalophilin and transcobalamin complexed with vitamin B-12 had molecular weights by gel filtration of 135 000 and 38 000 and by the formula of Svedberg 104 000 and 44 000, Stokes radii 4.97 nm and 2.65 nm, and sedimentation coefficients 5.39 S and 3.75 S, respectively. Electrofocusing resolved the cobalophilin complex into three main isoproteins isoelectric at pH 3.23, 3.42 and 3.69, and transcobalamin into only the main component isoelectric at a value as low as pH 3.47. Neither protein was capable of binding to the ileal intrinsic factor receptor.

Animals↗

Sequential affinity chromatography for the purification of antigens extracted from Onchocerca volvulus adult worms.

Adult Onchocerca volvulus worms were extracted sequentially with buffers of various ionic strengths. The extract was incubated with purified human onchocerciasis immunoglobulin-G (IgG) convalently coupled to Sepharose. Antigens were then eluted with 8 M urea and 7.5 M guanidine-HCl. IgG contained in the eluates was removed by incubating the eluates with rabbit anti-human IgG covalently coupled to Sepharose. As demonstrated by double immunodiffusion and "double" crossed immunoelectrophoresis at least three antigens were isolated. Most of the antigens were totally excluded from Sephadex G-200 but entered the included volume of Sepharose 6B. In electrofocusing they focused in acid regions. Antibodies were generated in rabbits against the three antigens isolated together. The antibodies were covalently coupled to Sepharose, which was subsequently used as a new affinity medium for the isolation of antigens. Such isolated antigens appeared to be identical with those with human onchocerciasis IgG. The isolated antigens and their corresponding artificial antibodies generated in rabbits were successfully applied in the enzyme linked immunoadsorbent assay. Host material and onchocerciasis IgG left behind during the purification procedure interfered in the assay system.

Antigens↗

Isolation of vitamin B-12-binding proteins by combined immuno and affinity chromatography. Comparative studies on the isolated and unisolated proteins.

Intrinsic factor or cobalophilin were removed by incubating human gastric juice and pig pyloric extract with purified anti-intrinsic factor and anti-cobalophilin immunoglobulin-G, respectively, covalently coupled to Sepharose. Cobalophilin (transcobalamin I) was also removed from pig serum either by using anti-cobalophilin immunoglobulin-G Sepharose or by Sephadex G-200 chromatography. The one remaining semipurified vitamin B-12-binding protein (intrinsic factor, cobalophilin or transcobalamin II) was then isolated by vitamin B-12-Sepharose affinity chromatography. Intrinsic factors, cobalophilins and transcobalamin II isolated by this two-step procedure were compared by double isotope techniques with the corresponding protein not subjected to affinity chromatography and found to be identical in reaction to antiserum, gel filtration and electrofocusing. The avidity of the isolated and unisolated intrinsic factors for the ileal intrinsic factor receptor was also the same.

Animals↗

Identification and characterization of intrinsic factor and cobalophilin from pig ileal and pyloric mucosa.

Pig ileal mucosa was found to bind about 240 ng vitamin B12/g and to contain two vitamin B12-binding proteins. One was highly active in the Schilling test, behaved immunologically as intrinsic factor and was responsible for about half of the total vitamin B12-binding capacity. The other binder was identified as cobalophilin (R-protein). Immunochemical purification of these proteins from pig ileum and pylorus was performed and the molecular characteristics (sedimentation and diffusion coefficients, Stokes radii, frictional ratios and molecular weights) of their vitamin B12 complexes were estimated. Isoelectric focusing revealed differences between the ileal and pyloric intrinsic factors but not between the cobalophilins. The mean isoelectric points of the pyloric and ileal intrinsic factors were pH 5979 and 5.30, respectively, whereas the corresponding figures for the cobalophilins were 4.13 and 4.10.

Animals↗

Vitamin B12.

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Absorption↗

Anti-nucleosome antibodies in the diagnosis of systemic lupus erythematosus.

OBJECTIVE: To study the prevalence and diagnostic significance of antibodies against nucleosomes in patients with systemic lupus erythematosus (SLE) as compared to five anti-nuclear antibody (ANA) assays. METHODS: The study included 305 patients with SLE, 125 patients with other autoimmune rheumatic diseases, and 415 healthy controls. Anti-nucleosome antibodies were measured by an enzyme-linked immunosorbent assay (ELISA) and ANA by immunofluorescence (IF) using Hep-2 cells. Anti-double-stranded DNA (anti-dsDNA) antibodies were measured by three commercial ELISAs and by IF using Crithidia luciliae as antigen. RESULTS: Compared to three ELISAs for anti-dsDNA, the anti-nucleosome assay was less sensitive (30% vs. 29-69%) but equally specific (90% vs. 77-95%) for SLE. The most sensitive test was ANA (76%), and the least sensitive was Crithidia (13%). The correlations between the different assays were good (p < 0.001 for all comparisons). CONCLUSION: The anti-nucleosome antibody assay does not offer additional information compared to conventionally used anti-dsDNA tests in the differential diagnosis of SLE.

Antibodies, Antinuclear↗

C3c-binding in inflammatory rheumatic diseases.

Sera from 73 patients, 24 with rheumatoid arthritis (RA), 10 with Sjögren's syndrome (SS, 4 with and six without RA), 17 with Reiter's syndrome (RS), 10 with systemic lupus erythematosus (SLE), 9 with Yersinia arthritis (YA), and 3 with mixed connective tissue disease (MCTD) were examined by enzyme-immunoassay (EIA) for the presence of C3c-binding IgG, IgM, and IgA activity (C3cBIgG, C3cBIgM, C3cBIgA). This activity probably consists of immunoconglutinins and immunoglobulin aggregates. Significantly elevated C3cBIgG was found in the sera of patients with RA (p less than 0.01), SS, and SLE (p less than 0.02). C3cBIgM was elevated in SS and SLE (p less than 0.02). C3cBIgA was increased in RA (p less than 0.005), in SS and YA (p less than 0.02). Correlations of C3c-binding with erythrocyte sedimentation rate, hemoglobin, C-reactive protein, IgG-binding onto platelets, serum IgG, IgM, and IgA levels, and with rheumatoid factors of IgM and IgA classes were computed. C3cBIgG, C3cBIgM, and C3cBIgA correlated significantly with each other, but usually not with other laboratory variables. The exceptions were positive correlations of C3cBIgM with IgM-rheumatoid factors in RA (p less than 0.05), and with serum IgM levels in the whole series of patients (p less than 0.05). The use of C3c represents a new principle to detect abnormalities in the sera of patients with inflammatory rheumatic diseases. However, the pathogenic significance of elevated C3c-binding remains unknown.

Complement C3↗