Humoral components of immunological response in nephrotic syndrome

Serum and urine were collected from 58 patients with nephrotic syndrome. Immunoglobulins (IgA, IgG and IgM), complement (C3) and transferrin levels were measured by single radial immunodiffusion. The extent of glomerular injury was estimated by determining the selectivity of proteinuria. The relationship between the severity of glomerular damage and serum concentrations of immunoglobulins and complement was assessed. Higher IgM and lower IgG serum concentrations were found in nephrotic patients than in normal controls (157 +/- 108 mg+ vs 127 +/- 38 mg% for IgM, 929 +/- 537 mg% for IgG). The difference was statistically significant (p less than 0.05 for IgM, p less than 0.001 for IgG). No correlation was present between the selectivity of proteinuria and serum levels of IgA, IgM, IgG or C3. The results indicate that abnormalities in humoral components of the immune system are present in nephrotic patients and are probably related to a basic immunological defect in the patients rather than to the severity of glomerular damage.     

Detection of antibodies to Angiostrongylus cantonensis in serum and cerebrospinal fluid of patients with eosinophilic meningitis

Adult and young adult antigens of Angiostrongylus cantonensis were purified by immuno-affinity chromatography and used to detect antibody in serum and cerebrospinal fluid (CSF), by enzyme-linked immunosorbent assay (ELISA), in cases of human eosinophilic meningitis or meningoencephalitis. The levels of IgG, IgA, IgM and IgE antibodies to A. cantonensis in these patients were higher than levels in control subjects. Antibodies in patients detected against adult and young adult worm antigens of A. cantonensis did not differ significantly. Significantly higher IgM and IgE antibody levels were observed in serum compared with CSF from infected patients (Student's t-test, P less than 0.05). Both adult and young adult A. cantonensis antigens proved to be highly sensitive in ELISA for serum antibodies; however, the sensitivity was significantly lower in tests on CSF.     

Immunobiochemical profiles of workers differing in the degree of occupational exposure to vinyl chloride

The immunobiochemical studies were conducted in a group of 98 production workers engaged in polyvinyl chloride manufacture from ethylene (group A workers) and in a group of 59 vinyl chloride workers from a chemical plant employing classic production technology from acetylene (group B workers). Both groups of workers were matched by age (group A workers: 37.7 +/- 8.66 years; group B workers: 34.9 +/- 11.2 years) and average exposure length (group A workers: 8.6 +/- 3.0 years; group B workers: 10.7 +/- 8.4 years). All workers were examined for the serum concentrations of immunoglobulins IgG, IgA and IgM and acute reactants lysozyme (LYS), transferrin (TRF), ceruloplasmin (CPL), alpha-l-antitrypsin (AlAT), alpha-2-macroglobulin (A2M) and orosomucoid (ORO). The statistical analysis included calculations of means, standard deviations and 95% confidence intervals. Differences in means were evaluated by t-test, differences in the distribution pattern of values by F-test. Abnormality of values was assessed by comparisons to normal values valid in Czechoslovakia. Group A worked in conditions meeting the MAC 10 mg VC.m-3 comparing with group B workers had elevated levels of IgG (P less than 0.005), IgA and IgM (P less than 0.001 both). Group B workers differed from group A workers by exhibiting significantly elevated levels of AlAT, and CPL. (P less than 0.001). The differences in the frequency of abnormal values between group A and group B worked in substantially less favourable hygienic conditions were significant for immunoglobulins elevated in group A and for ORO (P less than 0.01) and CPL (P less than 0.001) elevated in group B. The possible relationship of these immunobiochemical findings with the degree of vinyl chloride exposure are critically analyzed.     

Analysis of immunoglobulin,complements and CRP levels in serum of captive northern pig-tailed macaques (Macaca leonina)

The northern pig-tailed macaque(NPM,Macaca leonina) has become a widely used animal model in biomedical research. In this study, we measured serum immunoglobulin IgG, IgM, IgA, complement C3, C4 and CRP levels in 3-11 year old captive northern pig-tailed macaques using HITACHI 7600-20 automated chemistry analyzer in order to determine the influences of age and gender on these items. The results showed that serum IgA, IgM, C3 and C4 levels were not correlated with age(P0.05), while serum IgG levels increased progressively with age(r=0.202;P=0.045). Serum IgG, IgA, IgM and C3 levels were higher in females than in males(P0.05). Moreover, serum C3 concentration was both positively and strongly correlated with that of C4(r=0.700; P0.0001). This study provides basic serum immunoglobulin and complement data of captive northern pig-tailed macaques, which may prove useful for future breeding efforts and biomedical research.     

Detection of IgG and IgM antibodies with ELISA technique in human trichomoniasis

The direct wet mount examination of vaginal secretion, widely applied for the diagnosis of Trichomonas vaginalis infection in woman patients, is rapid and economical, however, the sensitivity of this technique is not so high. In this study enzyme-linked immunosorbent assay (ELISA) was employed for the detection of serum anti-T. vaginalis IgG and IgM antibodies from 30 vaginal trichomoniasis patients and 30 non-infected healthy persons. The results were as follows: 1. Serum ELISA-IgG value was 0.37 +/- 0.134 (Mean +/- S.D.) in vaginal trichomoniasis patients and 0.21 +/- 0.054 in healthy controls (p less than 0.005), and the sensitivity and specificity of ELISA for serum IgG antibody were 70.0% and 96.7%, respectively. 2. Serum ELISA-IgM value was 0.33 +/- 0.177 (Mean +/- S.D.) in vaginal trichomoniasis patients and 0.11 +/- 0.051 in healthy controls (p less than 0.005), and the sensitivity and specificity of ELISA for serum IgM antibody were 70.0% and 96.7%, respectively. 3. The ELISA-IgG values showed a significant correlation with ELISA-IgM values (r = 0.77, p less than 0.005). With above results, it is assumed that ELISA is a reliable method for the diagnosis of T. vaginalis infection and simultaneous measurement of serum IgG and IgM with this technique is recommended.     

Effect of cold exposure on the metabolism of immunoglobulins in rabbits.

The effect of low environmental temperature on the metabolism of IgG and IgM was examined in unimmunized rabbits. The half-lives of both IgG and IgM were less in animals kept at 4 degrees C for 6 weeks than in animals kept at 22 degrees C. Serum concentration of IgM and GG were unaltered by cold exposure but intravascular pool sizes tended to increase as a consequence of an expanded serum volume. Fractional turnover rates of both IgM and IgG were greater in cold-exposed animals. At both 4 degrees C and 22 degrees C, the fractional catabolic rate of IgM was independent of its serum concentration whereas that of IgG was correlated directly with its serum concentration. Absolute turnover of both IgM and IgG was accelerated by cold exposure. It is suggested that increased synthesis of immunoglobulin could account for the higher levels of antibody reportedly found in cold-exposed rabbits.     

IgG autoantibody activity in normal mouse serum is controlled by IgM

In the serum of normal BALB/c mice, IgG antibody reactivity to mouse actin and tubulin, DNA, and TNP groups was very low compared to that of the IgM. This activity was considerably increased when IgG was separated, by affinity chromatography on protein A-Sepharose, whereas no difference in the IgM activity was observed. Addition of IgM to IgG isolated from the same serum resulted in the inhibition of IgG binding to these Ag. Isolation of IgG antibodies on actin, TNP, and tubulin immunoadsorbents has indicated that at least part of the IgG antibodies is polyreactive. In order to understand this inhibition better, experiments with F(ab')2 fragments of IgG were performed. IgM inhibited the binding of F(ab')2 to the antigens in a dose-dependent manner and reacted with immobilized F(ab')2. IgM isolated on F(ab')2 immunoadsorbent, as compared to the initial IgM preparation, were less active toward the Ag but more inhibitory for IgG binding to the Ag. In some pathologic situations, IgM failed to inhibit some IgG antibody activities. The anti-DNA IgG activity from (NZB x NZW)F1 mice was not affected by autologous IgM. Similarly the anti-tubulin IgG from mice infected with Trypanosoma cruzi were less inhibited by IgM from autologous serum than antitubulin IgG from normal mice. These results are compatible with the existence in normal mice of an idiotypic-like network, regulating via an IgM population in the serum, the binding of IgG autoantibodies to self Ag. Modifications of this idiotype-anti-idiotype system might lead to the expression and/or expansion of autoreactive IgG-producing clones.     

Immunological profiles in workers of a power plant burning coal rich in arsenic content

A group of 47 male adults working in a thermal power plant burning coal containing 900 to 1,500 g of arsenic per ton dry weight was examined on the blood serum immunoglobulins IgG, IgA and IgM content and levels of acute reactants alpha-1-antitrypsin (A1AT), alpha-2-macroglobulin (A2M), transferrin (TRF), orosomucoid (ORO) ceruloplasmin (CPL), and lysozyme (LYS). Investigations in the control group comprising 27 workers from another power plant in the same district where the coal content of arsenic was more than 10 times lower were analogous. The inter-group differences in means were evaluated by t-test, differences in the association of values by F-test, and the correlations with age and the length of exposure were assessed using the regression analysis method. The differences in mean IgG, IgA, IgM, LYS and A2M levels between the exposed and control groups of workers were insignificant or of borderline significance only. In contrast, differences in TRF, ORO and particularly CPL levels were statistically highly significant, in all instances P less than 0.001. In the control group, persons with abnormal values in at least two immunobiochemical tests used accounted for 3.7%, in the group of the exposed for 51% (P less than 0.002). All these findings, especially the rise in CPL concentration levels in the exposed group are discussed on the background of the rise in cancer mortality rates found previously in this group of power plant workers.     

Early results of the studies of the effect of cimetidine on selected parameters of humoral and cellular immunity in patients with peptic ulcer

The study aimed at evaluating an early effect of cimetidine on the blood IgG, IgA, IgM, number and functioning of T-cells in peripheral blood of patients with ulcerative disease. The study involved 27 patients (9 women and 8 men), aged between 27 and 70 years (mean 52.6 +/- 10.9 years). Nine of these patients suffered from the peptic ulcer and 8--from duodenal ulcer. Cell-mediated and humoral immunity were evaluated simultaneously in all patients prior to and after a 4-week treatment with cimetidine administered orally in the dose of 200 mg four times daily. Rosette test with theophylline and leukocyte migration test were used to assess cell-mediated immunity. It was found that cimetidine significantly increases blood serum IgG (p less than .01), the number of theophylline-resistant lymphocytes in TRFC-TR (P less than 0.01), and T-cell response to higher mitogen concentrations (PHA and Con-A) (p less than .005 and p less than .001, respectively).     

The relationship between circulating CD5+ B lymphocytes and in vitro autoantibody synthesis in normal individuals.

Recent observations suggest that a subpopulation of B lymphocytes bearing the phenotype CD5+ may be enriched for cells committed to the synthesis and secretion of autoantibodies. We had previously shown that a subset of normal individuals has an expanded subpopulation of B lymphocytes committed to the synthesis of IgM and IgM-rheumatoid factor (RF), and that this condition was associated with HLA-DR4 (4). In these studies, we performed simultaneous quantitation of the size of the circulating CD5+ B lymphocyte subpopulation in a group of 20 normal donors, and of the pokeweed mitogen-induced in vitro synthesis of a panel of autoantibodies by the same peripheral blood cells depleted of CD8+ suppressor lymphocytes in 18 of the 20 individuals. The culture supernatants were assayed for total IgM and IgG, RF, IgM, and IgG anti-single-stranded DNA, anti-human thyroglobulin, and anti-tetanus toxoid. The mean percentage CD5+ B cells was 13.5 +/- 2.5%. There was no significant correlation between total B lymphocytes and CD5+ B cells (R = 0.25, P greater than 0.20. Positive correlations were found between the proportion of circulating CD5+ B lymphocytes and synthesis of RF (R = 0.73, P less than 0.001), and IgM anti-DNA (R = 0.58, P less than 0.03). Significant correlations were not found between CD5+ B cells and secreted IgM or IgG antibodies against the exogenous antigen tetanus toxoid, measured in the same supernatants. The antibodies produced in vitro by T cell-dependent B cell activation appear to have limited or no polyspecificity. These results indicate that the size of the circulating CD5+ B cell subpopulation in any given individual contributes importantly to the magnitude of autoantibody synthesis in cultures where T cell-mediated B lymphocyte activation takes place in the absence of suppressor signals.     

Amino acid composition of the mu chains of IgM of normal serum, monoclonal cryoglobulin and Waldenstrom macroglobulin

The purified IgM of normal human serum, monoclonal cryoglobulin and Waldenstrom macroglobulin were reduced, alkylated and subjected to Sephadex G-100 filtration. The mu chains separated were hydrolysed and analysed for amino acid content. In both monoclonal proteins the content of hydroxyproline in mu chains was lower than in the IgM of normal serum. The mu chain of monoclonal cryoglobulin contained less serine, methionine and tyrosine, whereas mu chains of Waldenstrom macroglobulin showed a lower level of cystine as compared with the IgM of normal serum. On the other hand, the content of valine and basic amino acids was higher in both monoclonal proteins.     

Effects of high dose progestin on serum lipids and lipid metabolizing enzymes in patients with endometrial cancer

The effect of high dose medroxyprogesterone acetate (MPA) on serum lipids, on adipose tissue lipoprotein lipase (LPL) and serum lecithin cholesterol acyltransferase activities were studied in 15 postmenopausal patients with endometrial cancer. After 2 weeks of MPA treatment total cholesterol decreased by 14% (P less than 0.001) and HDL cholesterol by 33% (P less than 0.01) from the respective pretreatment values; correspondingly the ratio of HDL to total cholesterol decreased (P less than 0.05). The decrease of HDL2 cholesterol was 35% (P less than 0.01) and that of HDL3 cholesterol 15% (P less than 0.01). The levels of serum triglycerides decreased significantly (P less than 0.05) during the treatment period. Serum LCAT activity was significantly lower (P less than 0.05) after treatment than before, but adipose tissue LPL activity was not altered. The mean serum testosterone level decreased significantly (P less than 0.001) from the pretreatment values. Significant positive correlations were present between LPL activity and MPA concentrations and between LPL activity and testosterone concentrations after the drug treatment.     

Serum immunoglobulin M (IgM) during early development of masu salmon (Oncorhynchus masou).

1. The development of serum immunoglobulin M (IgM) was studied by measuring IgM concentration in early developmental stages of masu salmon (Oncorhynchus masou) using single radial immunodiffusion. 2. IgM concentration was measurable from 88 days after hatching (46.5 micrograms/ml) and was maintained at a relatively constant level (less than 100 micrograms/ml) until 235 days after hatching. 3. IgM concentration increased significantly to a level of 691 +/- 37 micrograms/ml (mean +/- SE, N = 59) during the period from 251 to 429 days after hatching. The highest level of serum IgM was maintained from 429 days to 489 days after hatching. 4. The increase of IgM concentration was not accompanied by parallel increases in total serum protein concentration.     

Serum antibodies to Giardia lamblia by age in populations in Colorado and Thailand.

We measured levels of antibodies to Giardia lamblia by age in serum specimens from persons in Denver, Colorado, and Soongnern, Thailand. Serum levels of immunoglobulin (Ig) G, IgM, and IgA G lamblia-specific antibodies measured by enzyme-linked immunosorbent assay increased substantially during childhood in both geographic areas, although children in Soongnern showed significantly higher mean levels of each antibody class (P less than .05). After adolescence, levels of G lamblia-specific IgM fell steadily with age in both populations. In contrast, specific IgA levels remained elevated throughout life among the Thai but decreased to low levels among adults in Denver. Similarly, rates of carriage of G lamblia were high among children aged 1 to 4 years in Denver and Soongnern (14.3% versus 26.5%, respectively) but were much lower among adults in Denver (0% versus 14%; P less than .01). These data suggest that levels of G lamblia-specific IgM may reflect exposure to the parasite early in life in both areas. Levels of parasite-specific IgA may reflect recurrent exposure to G lamblia in Soongnern, where G lamblia is endemic, but less frequent exposure to the parasite in Denver, where exposure is often episodic.     

Studies on the neutralization of herpes simplex virus. XI. Differences between slow-reacting complement-requiring neutralizing (s-CRN) antibodies in IgG and IgM

Late IgG and IgM from a rabbit immunized with herpes virus were tested for ordinary neutralizing (N) antibody, complement-requiring neutralizing (CRN) antibody and in addition CRN antibody detectable by overnight sensitization at 0 C (s-CRN antibody). Heat stability tests showed that IgG s-CRN antibody was slightly less resistant to heating at 70 C than were N and CRN antibodies, whereas all three activities of IgM were quickly degraded at this temperature. Dose-response curves with varying amounts of complement (C) or anti-antibody revealed a marked difference between IgG s-CRN and IgM s-CRN antibodies. While 1-hr sensitization at 37 C was insufficient to detect IgG s-CRN antibody, it had the same effect as overnight sensitization at 0 C for IgM s-CRN antibody. When sensitization at 0 C was prolonged to 3 days, unexpectedly high endpoints exceeding 1:10,000 were obtained even with IgM. consequently, enhancement by C was several hundred-fold with IgM in contrast to 5- to 10-fold enhancement of IgG s-CRN antibody, which was similar to that after overnight sensitization. Also IgM obviously required more C than did IgG. These results suggest that IgM of late serum is slower reacting and more C-dependent than IgG s-CRN antibody. Tests with early rabbit sera indicated that the s-CRN antibody detectable by 3-day sensitization reaches a high level before the appearance of N antibody.     

The relationship of serum vitamin A, cholesterol, and triglycerides to the incidence of ovarian cancer

Prospective and retrospective studies have suggested that serum vitamin A and total cholesterol levels may be associated with cancer. Our study showed that the mean (+/- SEM) concentrations of serum vitamin A 489 +/- 33.28 (mean +/- SEM micrograms/liter and serum total cholesterol 174.7 +/- 8.96 (mean +/- SEM) mg/dl from ovarian cancer patients in Singapore were significantly lower than the respective values of 668 +/- 25.10 (mean +/- SEM) microgram/liter and 210.7 4.48 (mean +/- SEM) mg/dl from noncancerous control subjects (P less than 0.0001 for both compounds). In addition, ovarian cancer patients did not show significantly lower serum triglyceride levels than the control subjects. Age did not significantly correlate the serum vitamin A and total cholesterol concentrations, but there was correlation with respect to the serum triglyceride levels. There were moderate correlations between vitamin A and cholesterol levels (r = 0.36, P less than 0.0027) and between cholesterol and triglyceride levels (r = 0.37, P less than 0.0024) in the control subjects but not in the cancer patients. Vitamin A levels correlated moderately with triglyceride levels in both the cancer patients (r = 0.42, P less than 0.0258) and the control subjects (r = 0.33, P less than 0.0069). The inverse relationship between the incidence of ovarian cancer and serum vitamin A and serum total cholesterol concentrations may have distinct implications for preventive medicine and public health.     

Serum transferrin as a marker of bone growth in boys: correlation with serum alkaline phosphatase activity, plasma insulin-like growth factor 1 and rate of growth in height

Transferrin (TF) has a growth promoting activity in cell culture. The aim of this work was to study possible relationships between serum TF, alkaline phosphatase activity (ALP), plasma insulin-like growth factor 1 (IGF-1) levels and rate of height increase in boys. 149 boys aged 13-15 yrs were studied. TF levels were measured using turbidimetric method. The serum levels of ALP could be used as a biochemical marker for bone formation. Significant correlation was found between serum TF levels and ALP levels (r = 0.31, P less than 0.0005). The TF levels are higher in iron-deficiency anemia. The hemoglobin (Hb) and serum ferritin were measured in all boys. Thirty-one of 149 boys had no iron-deficiency anemia (Hb 14.0 g/dl and serum ferritin 23 ng/ml). The rate of growth in height was estimated over a 5 month period. In these boys, the rate of growth in height was significantly correlated with serum TF levels (r = 0.37; P less than 0.05). A significant correlation was found between serum TF levels and plasma IGF-1 levels (r = 0.45; P less than 0.05). These data indicate that serum TF levels may be a marker of skeletal growth in normal boys.     

Relation of human serum antibody against Staphylococcus epidermidis cell surface polysaccharide detected by enzyme-linked immunosorbent assay to passive protection in the mouse

A specific and rapid enzyme-linked immunosorbent assay (ELISA) has been applied for the detection of immunoglobulins to Staphylococcus epidermidis cell surface polysaccharides in human serum. Positive IgG, IgM and IgA titres of more than 1:6400, 1:1600 and 1:400 were observed with this assay against passive protective human serum. However, IgG, IgM and IgA titres of less than 1:400, 1:100 and 1:50 were shown in non-protective serum. When the cross-reactivity of passive protective human serum to homologous and heterologous cell surface polysaccharides was examined by inhibition test with ELISA, remarkable inhibition was shown with homologous cell surface polysaccharide, whereas no inhibition was observed with heterologous substances. According to these results, the quantitation of human serum antibody by the ELISA method against Staph. epidemidis cell surface polysaccharide was found to be significant for the demonstration of passive protective activities against Staph. epidermidis.     

Periovulatory changes in serum concentration and ovarian content of 5 alpha-androstane-3 alpha, 17 beta-diol in the adult rat

The high amounts of 5 alpha-androstane-3 alpha, 17 beta-diol (3 alpha-diol) present in immature female rats decline towards first ovulation, but on the day of first proestrus a peak is seen. This raises the possibility that during adulthood similar proestrous peaks may occur. Therefore, serum concentrations and ovarian content of 3 alpha-diol were estimated every two hours between 0900 and 2100 h in adult cyclic rats on the day of proestrus. In the same rats, serum concentrations of estradiol (E2), progesterone (P) and luteinizing hormone (LH) were measured, as were ovarian contents of E2 and P. A significant elevation in ovarian 3 alpha-diol was found between 0900 and 1700 h proestrus, whereas serum concentrations of 3 alpha-diol were elevated from 1300 to 2100 h. The high morning values of ovarian 3 alpha-diol correlated with those for ovarian E2 (p less than 0.005); the elevated serum concentrations of 3 alpha-diol during the afternoon correlated with serum P (p less than 0.005) and with serum LH concentrations (p less than 0.005). Serum and ovarian values were positively correlated for P and E2, but not for 3 alpha-diol. The rise in serum 3 alpha-diol could be prevented by blocking the LH surge with sodium pentobarbitone (Nembutal; 35 mg/kg b.w.) administered at 1300 h. In Nembutal-treated rats, the concentration of 3 alpha-diol at 1700 h (886 pg/ml) was significantly lower than in saline-treated control rats (1135 pg/ml; p less than 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)     

Prophage type of recombinants produced by cell fusion between various combinations of lysogenic or non-lysogenic substrains from two Staphylococcus aureus L-forms

Streptomycin (SM)- or erythromycin (EM)-resistant lysogenic and non-lysogenic substrains were produced from two Staphylococcus aureus L-form strains lysogenic for different prophages, namely, EMT-L (prophage alpha) and 209P (prophage beta). Cells of these L-form substrains were fused in various combinations using polyethylene glycol (PEG), and the frequency of recombinants selected as double resistance to both SM and EM and the prophage types of these recombinants were examined. In all the combinations, the frequency of recombinants was greater when the cells were treated with PEG than when they were not, and the difference was statistically significant (p less than 0.01) in 13 combinations. Combination between the lysogenic SM-resistant EMT-L substrain [EMT(Smr-alpha)] and lysogenic EM-resistant 209P-L substrain [209P(Emr-beta)] and the reverse combination, between 209P(Smr-beta) and EMT(Emr-alpha), resulted in a majority of recombinants harboring prophage beta. The former combination yielded recombinants that all held both prophage alpha and beta.