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Serum levels of soluble IL-2R, IL-4, and soluble Fc epsilon RII in adult bronchial asthma.

Examination was made of the serum levels of interleukin 4 (IL-4), soluble Fc epsilon RII (sFc epsilon RII), and soluble interleukin 2 receptor (sIL-2R) in 77 adult patients with bronchial asthma. All the patients had mild to moderate asthma and their symptoms were well controlled. The results were compared with values for 75 control subjects to clarify the involvement of these factors in the pathogenesis of bronchial asthma. Serum sIL-2R was elevated in asthmatics compared with control subjects (459.0 +/- 30.4 U/ml vs 251.5 +/- 10.0 U/ml; p < 0.001). The IL-4 and sFc epsilon RII levels were also elevated beyond those in the controls (IL-4: 1.89 +/- 0.13 pg/ml vs 1.08 +/- 0.15 pg/ml; p < 0.001) (sFc epsilon RII: 313.2 +/- 18.8 U/ml vs 228.8 +/- 6.7 U/ml, p < 0.001). A weak but significant correlation was observed between sIL-2R and sFc epsilon RII (r = 0.46, p < 0.001). Correlation among other parameters was not found. The patients were divided into atopic and nonatopic groups, based on the presence or absence of positive reaction to skin prick tests using extracts of common aeroallergens and positive specific IgE against house dust mite (Dermatophagoides pteronyssinus [Dp]). Mean IL-4 in the atopic group, with positive skin reaction and > 0.7 kU/L specific IgE against Dp was significantly elevated compared with the nonatopic group (2.35 +/- 0.26 pg/ml vs 1.56 +/- 0.12 pg/ml; p < 0.005). Such differences could not be detected in sIL-2R, sFc epsilon RII levels among the two groups. No significant correlation could be found among IL-4, sFc epsilon RII, and total IgE level. The activation of T cells and B cells would thus appear essential to the pathogenesis of bronchial asthma and the hypothesis that atopic status is associated with the preferential activation of TH2 cells which selectively produce IL-4 could be supported. The regulatory mechanism of IgE synthesis would not appear to be the only responsible factor.

Adult↗

The cyanogen bromide peptides of bovine soluble and insoluble collagens. I. Characterization of peptides from soluble type I collagen by sodium dodecylsulphate polyacrylamide gel electrophoresis.

Acid soluble collagen and purified alpha1 and alpha2 chains were prepared from bovine skin and digested with cyanogen bromide. The resultant peptides were separated by electrophoresis on polyacrylamide gels containing sodium dodecylsulphate. Thirteen peptides obtained from the alpha1 chain and two from the alpha2 chain were identified as overlapping sequences containing methionine-derived peptide bonds not cleaved during the reaction. Those uncleaved peptides from the alpha1 chain accounted for approximately 30% of the total digest, as determined by planimetry on the staining profiles, and their relative proportions indicated that efficiency of cleavage at specific methionine residues was dependent upon position in the sequence. Alpha1CB5-8 was most resistant to cleavage while alpha1CB0,1-2 and alpha1CB7-6 were most susceptible. Peptides of a 2:1 (w/w) mixture of alpha1 and alpha2 chains and of the acid-soluble collagen gave electrophoretograms which were essentially a summation of those obtained from the individual chains.

Animals↗

Serum levels of soluble interleukin 6 receptor and soluble gp130 are elevated in patients with localized scleroderma.

OBJECTIVE: To investigate the clinical relevance of serum soluble interleukin 6 receptor (sIL-6R) and soluble gp130 (sgp130) in localized scleroderma. METHODS: Serum levels of sIL-6R and sgp130 were examined by ELISA in 45 patients with localized scleroderma. Twenty patients with systemic sclerosis (SSc) and 20 healthy individuals served as controls. RESULTS: Serum levels of both sIL-6R and sgp130 were significantly elevated in patients with localized scleroderma compared with healthy controls. Moreover, serum sgp130 levels in patients with localized scleroderma were significantly higher than in patients with SSc. In patients with localized scleroderma, elevated sIL-6R levels significantly correlated with levels of IgM antihistone antibodies, the presence of rheumatoid factor, the number of linear lesions, and the number of body areas involved. Elevated sgp130 levels were significantly associated with levels of IgG antihistone antibodies, the number of plaque lesions, the total number of lesions, and the number of body areas involved. CONCLUSION: These results suggest that elevated serum sIL-6R and sgp 30 may reflect activation of the IL-6 system that may be associated with the development of sclerotic lesions and autoantibody production in localized scleroderma.

Adolescent↗

Elevation of serum eosinophil cationic protein, soluble tumor necrosis factor receptors and soluble intercellular adhesion molecule-1 levels in acute bronchial asthma.

Asthma is a disease characterized by chronic airway inflammation. Many inflammatory cells and molecules contribute to its pathogenesis. Soluble tumor necrosis factor receptors (sTNFRs) and soluble intercellular adhesion molecule-1 (sICAM-1) play regulatory roles in the inflammation. But their roles in the inflammation of asthma have not been well defined. This study was done to examine the changes in serum levels of these molecules in acute asthmatic patients. The concentrations of eosinophilic cationic protein (ECP), sTNF-RI, sTNF-RII, and sICAM-1 were measured in sera of 24 asthmatic patients during acute attack, in 18 patients at 24 h, and in 10 patients at 7 days after attack and in sera of 14 healthy control subjects by ELISA method. Serum levels of ECP, sTNF-RI, sTNF-RII and sICAM-1 in the patients with asthma during attack were significantly higher than those of the controls (p < 0.001, p < 0.01, p < 0.05, p < 0.05 respectively) and stayed high up to the 7th day. In conclusion, high serum levels of sTNF-RI, sTNF-RII and sICAM-1 suggest that these molecules may contribute to the regulation of allergic inflammation and may reflect the severity of inflammation in the airway of asthmatic patients.

Acute Disease↗

Soluble interleukin-2 receptor and soluble CD8 molecules in cerebrospinal fluid and serum of patients with multiple sclerosis.

The purpose of this study was to analyse soluble interleukin-2 receptor (sIL-2R) and soluble CD8 (sCD8) molecules in the cerebrospinal fluid (CSF) and serum of 18 patients with definite multiple sclerosis (MS) and of 16 with noninflammatory neurologic diseases (NIND). All MS patients suffered from an exacerbation of the relapsing-remitting form of the disease within one month before examination. The mean serum levels of sIL-2R and sCD8 in the MS patients were not significantly different from those of NIND patients. Only one patient with MS had detectable sIL-2R in the CSF. CSF sCD8 was detectable in 10 of 18 MS patients and in 1 of 16 NIND patients. Our data indicate that the CSF and serum sIL-2R concentrations do not correlate with the disease activity. Conversely, increased levels of sCD8 only in the CSF of MS patients support the hypothesis of an intrathecal activation of CD8+ cells in MS. We think that CSF sCD8 can be a useful marker for the presence of activated T cells in the central nervous system.

Adult↗

The evaluation of soluble Fas and soluble Fas ligand levels of bronchoalveolar lavage fluid in lung cancer patients.

Fas-Fas Ligand (FasL) is one of the major mediator system that activates programmed cell death. Cleavage of membranebound FasL by a metalloproteinase-like enzyme resulted in the formation of soluble FasL (sFasL). sFasL as well as the transmembrane form of FasL binds to Fas and transduces apoptotic signal in Fas-expressing cells. It's suggested that soluble Fas (sFas) and sFasL has an impact on tumor progress and immune escape feature of tumor cells from the host immune system. Since Fas antigen expression in the lungs has been localized to alveolar and bronchial epithelial cells, in this study we aimed to investigate the sFas (pg/mL) and sFasL levels (pg/mL) of bronchoalveolar lavage (BAL) fluid in lung cancer patients. Study population was consisted of 27 patients with lung cancer (mean age 62.9 +/- 10.7 years, 25 control subjects (mean age 47.9 +/- 13.9 years). BAL was performed under local anesthesia, on the unaffected lung of patients; either subsegments of right middle or lingula. BAL sFas and sFasL were evaluated by using ELISA method. The mean levels of sFas was 60.8 +/- 56.8 in lung cancer patient and 39.5 +/- 25.9 in control subjects (p> 0.05). The mean levels of sFasL was 51.6 +/- 39.2 in cancer patient and 41.2 +/- 27.4 in control subjects (p> 0.05). In conclusion, although we did not observe any significant difference between two groups, higher BAL levels of sFas and sFasL levels in lung cancer patients than control subjects, made us thought that apoptosis might have a role development and progression of lung cancer.

Adult↗

[Levels and clinic significance of serum soluble Fas and soluble Fas ligand in coal workers' pneumoconiosis].

OBJECTIVE: To investigate the changes of levels and clinic significance of serum soluble Fas (sFas) and soluble FasL (sFasL) in coal workers' pneumonoconiosis. METHODS: Serum levels of sFas and sFasL were determined in 52 patients with silicosis, 57 coal workers' pneumonoconiosis, 46 healthy underground coal workers' (the underground control group) and 40 healthy volunteers working on the ground (the ground control group) with a sandwich ELISA. RESULTS: Compared to the underground control and the ground control group, the serum levels of sFas and sFasL in the patients with silicosis and the coal workers' pneumonoconiosis were significantly higher (P < 0.01). Serum levels of sFas and sFasL in the underground control group were significantly higher than those in the ground control group (P < 0.01); Serum sFas levels in coal workers' pneumonoconiosis was significantly higher than those in the patients with silicosis (P < 0.01). Although the serum sFasL levels was also increased, there was no significant difference (P > 0.05). In the patients with silicosis and the coal workers' pneumonoconiosis patients, the serum sFas levels in Phase I patients combined with emphysema and simple Phase II + III patients were significantly higher than those in simple Phase I patients (P < 0.01). There was no significant difference in the serum sFasL levels among various groups with different parameters of pneumonoconiosis. In the patients with silicosis and the coal workers' pneumonoconiosis, serum levels of sFas and sFasL were not significantly altered among different duration of exposure to dusts. There was no correlation between serum levels of sFas and sFasL in the patients with silicosis while there was a slightly positive correlation between sFas and sFasL levels in the coal workers' pneumonoconiosis (r = 0.479, P < 0.05). CONCLUSION: In the patients with silicosis and the coal workers' pneumonoconiosis, the serum levels of sFas and sFasL are abnormal and associated with the development of the pneumonoconiosis. The changes of serum sFas levels may indicate the development and progression of the pneumonoconiosis. The detection of the serum sFas level may be used in the differential diagnosis for the silicosis and the coal worker's pneumonoconiosis.

Adult↗

[Change in serum levels of soluble Fas and soluble FasL in patients with multiple organ dysfunction syndrome].

OBJECTIVE: To evaluate the significance of changes in the serum levels of soluble Fas (sFas) and soluble Fas ligand (sFasL) in patients with multiple organ dysfunction syndrome (MODS) regarding its diagnosis and judgement of severity and outcome, and to investigate the correlations between the levels of tumor necrosis factor-alpha (TNF-alpha), sFas and sFasL. METHODS: Enzyme linked immunoadsorbent assay (ELISA) was used in the determination of serum sFas, sFasL and TNF-alpha in 36 patients with MODS. Thirty-two non SIRS patients and 20 healthy individuals comprised the control groups. The acute physiology and chronic health evaluation III (APACHE III) scoring system and modified MODS scoring system were used to assess patients' clinical severity. The differences of sFas and sFasL levels between MODS group and control groups and between survival and dead patients were observed. The correlations between sFas, sFasL and TNF-alpha levels and severity of MODS and the correlations between the TNF-alpha levels and the levels of sFas and sFasL were also observed. RESULTS: The serum levels of sFas, sFasL and TNF-alpha in patients with MODS were significantly higher than those in controls (P<0.05 or P<0.01), and were associated with severity of the disease (all P<0.01) . The sFas and sFasL levels were found to be significantly higher in the patients who eventually died as compared to those in the patients who survived (both P<0.05). Positive correlations were noted between the TNF-alpha levels and the levels of sFas and sFasL(both P<0.01). The serum levels of sFas and sFasL were elevated with the increase of the number of failure organs in MODS patients. CONCLUSION: The serum levels of sFas and sFasL may serve as diagnostic and prognostic indicators for MODS. TNF-alpha may help enhance the expression of Fas/FasL system.

Adolescent↗

[Soluble high molecular weight derivatives of pancreatic trypsin inhibitor. Modification of pancreatic trypsin inhibitor by soluble polysaccharides activated by titanium tetrachloride].

The activation of soluble polysaccharide matrices by titan tetrachloride was studied. A method for obtaining soluble preparations of pancreatic trypsin inhibitor bound to the activated polysaccharide matrix completely retaining its inhibiting activity was developed. It was shown that the equilibrium and kinetic constants for association of modified preparations of pancreatic inhibitor with commercial trypsin did not practically differ from those for native inhibitor. It was found that modification by titan tetrachloride-activated polysaccharides stabilized the protein molecule.

Kinetics↗

[Changes in water-soluble, urea-soluble and membrane intrinsic proteins in human senile cataract].

Gel filtration of water-soluble protein shows a substantial increase in HM+ alpha-crystallin and a marked decrease in beta- and gamma-crystallins in cortical cataract. A decrease in beta 1-crystallin in cortical punctate opaque lenses is also striking. In nuclear cataractous lenses HM+ alpha- and beta-crystallin increase, while gamma-crystallin decreases. The urea-soluble protein from clear lenses contains mainly of alpha beta chain, whereas in cataractous lenses the relative amounts of the 28 and 23ku polypeptides (the components of beta-crystallin) increased markedly. In cataractous lenses the relative amount of membrane intrinsic proteins decreases slightly and it has little statistical meaning.

Cataract↗

Serum concentrations of neopterin, soluble interleukin 2 receptor, and soluble tumor necrosis factor receptor in Wegener's granulomatosis.

OBJECTIVE: To investigate the possible relationship between serum levels of neopterin, soluble tumor necrosis factor-55 receptor (sTNF-55R), and soluble interleukin 2 receptor (sIL-2R) with disease activity in patients with Wegener's granulomatosis (WG). METHODS: Serum neopterin was measured by radioimmunoassay, sTNF-55R and sIL-2R were measured by ELISA in 26 patients with WG. RESULTS: Serum neopterin, sTNF-55R, and sIL-2R were significantly elevated in patients with generalized WG compared with healthy controls. Concentrations of the analytes correlated with disease activity indices in patients without infectious complications. The highest elevations of all 3 variables were observed in patients with intercurrent infections. CONCLUSION: The increased serum levels of neopterin, sTNF-55R, and sIL-2R suggest activation of cellular immunity in WG.

Adult↗

High levels of TNF, soluble TNF receptors, soluble ICAM-1, and IFN-gamma, but low levels of IL-5, are associated with hepatosplenic disease in human schistosomiasis mansoni.

In a case-control study based in two areas of Kenya, hepatosplenic schistosomiasis mansoni was shown to be linked with low levels of IL-5 and with correspondingly high IFN-gamma, TNF, and circulating soluble TNF receptor I (sTNFR-I), sTNFR-II, and sICAM-1. PBMC from the hepatosplenic cases responded to in vitro Ag stimulation with significantly higher levels of IFN-gamma and TNF, but lower levels of IL-5, compared with nonhepatosplenic controls matched for age and infection intensity. Most of these correlations were confounded by differences between geographical areas. However, principle component analysis identified a high IFN-gamma and TNF, and low IL-5 axis in the data as the first principle component; this was significantly associated with hepatosplenomegaly (p < 0.0005) even after controlling for area. High plasma levels of sTNFR-I (p < 0.001), sTNFR-II, (p < 0.0001), and sICAM-1 (p < 0.009) were also significantly associated with hepatosplenomegaly, independently of area, in the case of the soluble forms of both TNF receptors. These parameters were negatively related to IL-5. These results suggest that proinflammatory cytokines are involved in the hepatosplenic disease process in infected individuals who have low anti-inflammatory Th2 responses and that sTNFR may be a useful circulating marker for this disease process, perhaps reflecting the level of TNF activity in hepatic tissues.

Adolescent↗

Serum levels of interleukin 6 (IL-6), oncostatin M, soluble IL-6 receptor, and soluble gp130 in patients with systemic sclerosis.

OBJECTIVE: To determine whether serum interleukin 6 (IL-6), oncostatin M (OSM), soluble IL-6 receptor (sIL-6R), and soluble gp130 (sgp130) levels in patients with systemic sclerosis (SSc) are elevated and whether they are correlated with the clinical or serological features of the disease. METHODS: Serum samples from patients with SSc (n = 55) and control subjects (n = 20) were investigated by ELISA. Patients were divided into 4 groups: 12 with limited cutaneous SSc (lSSc) < or = 3 years' duration (early lSSc), 22 with lSSc > 3 years' duration (late lSSc), 9 with diffuse cutaneous SSc (dSSc) < or = 3 years' duration (early dSSc), and 12 with dSSc > 3 years' duration (late dSSc). RESULTS: Serum IL-6 levels were significantly elevated in patients with early dSSc compared with controls. In addition, serum IL-6 was detected more frequently in patients with pulmonary fibrosis, and these values were inversely correlated with the percentage of vital capacity of individual patients. Furthermore, serum IL-6 levels were correlated with erythrocyte sedimentation rates, C-reactive protein, and IgG and IgA levels in patients. Serum sIL-6R levels were significantly higher in patients with lSSc versus controls. Serum OSM and sgp130 levels were not significantly elevated in patients with SSc compared with controls. CONCLUSION: We suggest that IL-6 and sIL-6R may contribute to the disease process in SSc. In particular, IL-6 may be related to the early phase of the disease and the development of pulmonary fibrosis in SSc.

Adolescent↗

Solubility in binary solvent systems: 8. Estimation of binary alkane plus p-dioxane solvent nonideality from measured anthracene solubilities.

Experimental solubilities are reported for anthracene in binary solvent mixtures containing p-dioxane with n-hexane, cyclohexane, n-heptane, methylcyclohexane, n-octane, cyclooctane, and isooctane at 25 degrees C. Results of these measurements, used in conjunction with the nearly ideal binary solvent (NIBS) model, enabled excess Gibbs free energies, delta GBCfh, of the seven binary solvent mixtures to be estimated. Estimated values for p-dioxane plus cyclohexane, p-dioxane plus n-heptane, and p-dioxane plus methylcyclohexane mixtures are in reasonable agreement with published values based on vapor pressure measurements.

Alkanes↗

Direct evidence for a soluble methane monooxygenase from type I methanotrophic bacteria: purification and properties of a soluble methane monooxygenase from Methylomonas sp. GYJ3.

The hydroxylase and reductase components of a soluble methane monooxygenase from type I methanotrophs--Methylomonas sp. GYJ3--were purified by a multiple-step LC procedure. The hydroxylase (approximately 240 kDa, determined by an HPLC-size exclusion chromatography method) has three subunits with molecular masses of 56, 43, and 27 kDa, suggesting that the enzyme has an (alphabeta gamma)2 subunit structure. The HPLC method was developed to purify the hydroxylase component, and the purified protein has a specific activity of 541 nmol propene oxide x mg(-1) protein x min(-1), which is two times the specific activity of the protein purified by the two-step LC procedure. The iron content in the hydroxylase purified by the two-step LC procedure is 2.1 mol of Fe per mole of protein, but the iron content in the protein by the HPLC procedure is 3.78 mol of Fe per mole of protein. The diversity of iron contents in this protein is due mainly to the use of different purification methods. The reductase has a molecular mass of 42 kDa. The UV-VIS spectrum of the protein is similar to that of proteins from other methanotrophs, suggesting that the protein contains a FAD cofactor and a [2Fe-2S] center. The partially purified component B stimulated the MMO activity of the hydroxylase and reductase system by 40-fold.

Chromatography, High Pressure Liquid↗

Presence of mRNAs encoding the soluble D-factor/LIF receptor in human choriocarcinoma cells and production of the soluble receptor.

Four different kinds of cDNAs encoding the receptor for the differentiation stimulating factor (D-factor)/leukemia inhibitory factor (LIF) were isolated from a human liver cDNA library. Three kinds of the cDNAs had different intronic sequences, which introduced a termination codon before the transmembrane domain. However, the site of insertion in human cDNAs is different from those in mouse cDNAs. Recombinant proteins produced by COS-7 cells after transfection of these cDNAs bound D-factor/LIF. mRNAs corresponding to these cDNAs are expressed by the human liver, placenta, and choriocarcinoma cell line NJG. In conditioned medium of choriocarcinoma cells, a native soluble receptor for the D-factor/LIF with a dissociation constant of 7.8 nM and a MW of 160 kDa was found.

Alternative Splicing↗

Overexpression, purification, and use of a soluble human interleukin-4 receptor alpha-chain/Ig gamma 1 fusion protein for ligand binding studies. Characterization of ligand binding to soluble IL-4 receptor alpha-chain by surface plasmon resonance measurements and by microtiter-plate-based ELISA with biotinylated IL-4.

The pleiotropic cytokine IL-4 transmits cellular signals mainly via the IL-4 receptor complex, with the alpha-chain as the high affinity binding subunit. Here we describe the overexpression of a soluble IL-4R alpha-chain (sIL-4R) as a fusion to immunoglobulin gamma 1 heavy chain, consisting of the H-CH2-CH3 domains, in baby hamster kidney cells. The dimeric fusion protein named sIL-4R:E gamma 1 was purified from culture supernatant by protein-A affinity chromatography, yielding up to 10 mg/l homogenous protein which was highly stable. The antibody-like features of the sIL-4R:E gamma 1 fusion protein allowed immobilization on a biosensor matrix for surface plasmon resonance measurements by direct amine coupling as well as immobilization on microtiter plates coated with protein A for displacement binding. Kinetic parameters (kon and koff) for binding of IL-4 or the antagonistic mutant IL-4(Y124D) to the sIL-4R:E gamma 1 fusion protein on the chip as determined with the BIAcore instrument showed a high affinity binding with KD = 239 +/- 35 pM and KD=148 +/- 33 pM, respectively. The extremely high kon rate and the relatively slow koff rate for both ligands highlighted the limits of the BIAcore technology. The binding affinity as calculated in displacement binding studies with biotinylated IL-4 was similar for IL-4 and IL-4(Y124D) (IC50=1.1nM), thus offering a simple alternative for initial characterization of IL-4 mutants.

Animals↗

Soluble Fc epsilon RII/CD23 in patients with autoimmune diseases and Epstein-Barr virus-related disorders: analysis by ELISA for soluble Fc epsilon RII/CD23.

The low-affinity Fc receptor for IgE (Fc epsilon RII/CD23) and its soluble form (sCD23, IgE-binding factor) have multiple functions, and enhanced levels of these are associated with various immunological diseases. We established two sensitive ELISA systems using enzyme-conjugated mAb and biotinylated mAb. The detection limits of the ELISA systems were 0.03 and 1.0 ng/ml, which showed good correlation in the range 1.0-10 ng/ml. In the ELISA system using enzyme-conjugated mAb, the average sCD23 concentration in 303 normal healthy volunteers was 1.4 +/- 0.3 ng/ml. In the ELISA system using biotinylated mAb, sCD23 levels in normal healthy volunteers showed almost the same values. In patients with autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, Sjögren syndrome, progressive systemic sclerosis, and mixed connective tissue disease, the sCD23 levels were significantly higher than those in normal individuals. Furthermore, in Epstein-Barr virus-related disorders after liver transplantation with immunosuppression, plasma levels of sCD23 rapidly increased to more than 12 ng/ml when clinical symptoms were evident. In addition, the sCD23 values remained high, although elevated GOT levels gradually decreased to standard values and EBV hepatitis improved. These data suggest that sCD23 levels are a sensitive marker of autoimmune diseases and EBV-related disorders in addition to allergic disorders. The ELISA system for sCD23 may be an additional diagnostic tool in estimating the clinical courses of these diseases.

Adult↗