Immunologic aspects of term pregnancy toxemia. A study of immunoglobins and complement

IgA, IgG, and IgM were measured by the single radial immunodiffusion method in umbilical cord and maternal sera in toxemia patients and in normal term pregnant cases. Complement (C' 3) levels were determined only in maternal sera. Quantitation of IgA, IgG, and IgM was performed in concentrated serial urine samples from three eclamptic and three normal control patients. Results show that there are lower IgG and IgM levels in the sera of mothers with toxemia of pregnancy; however, paradoxically higher IgM levels were detected in cord sera from their newborn infants without evidence of placental leakage or increased neonatal infection. The finding of the low IgG and IgM levels appears to be due in part to immunoglobin loss into the urine. The unchanged complement levels in toxemia, in this study, are not suggestive of an active immunologic process, but the high IgM in the newborn infants of toxemic mothers is unexplained and may represent active immunologic disease. Clearly, investigations of tissue binding and the formation of immunoglobin complexes should be carried out to better explain these abnormal findings.     

Changes in serum immunoglobulin levels during phenytoin treatment of epilepsy

Immunoglubulin concentrations were determined by radial immunodiffusion in sera from 15 epileptic patients before and during phenytoin therapy. Three reaction patterns were recorded: Two patients developed IgA deficiency (less than 0.05 mg/ml) during the first 3-4 months of treatment. Both patients also had a decrease in serum IgG and IgM, but no significant fall or increase in serum IgE. The IgA deficiency state was apparently reversible, since normalization of serum levels occurred after withdrawal of phenytoin. Five patients developed a 35-80 per cent reduction in serum IgA. In these patients, the decline in serum levels of IgG and IgM was inconsistent. Eight patients showed no significant fluctuations in serum immunoglobulins during phenytoin treatment. When a fall in serum IgA occurred, it did not correspond to a fall in serum or in red cell folate. Mean serum IgG was lower (9.37 mg/ml) in epileptic patients who had taken phenytoin for less than 1 year and had a low IgA, than in patients who had taken phenytoin for 10 years or more (11.50 mg/ml).     

Study on the determination of IgA, IgG, IgM, a1-antitrypsin, haptoglobin and transferrin with the kinetic nephelometric method. Comparison with the radial immunodiffusion method (author's transl)

The concentration of IgA, IgG, IgM, a1-antitrypsin, haptoglobin and transferrin was determined by the kinetic nephelometric method (Beckman immunochemistry analyzer) and by the radial immunodiffusion method (Behring partigen immunodiffusion plates). The investigation was carried out in 151 normal and pathological sera. The results obtained with the radial immunodiffusion method were for IgA, IgG and IgM in middle about 15%, for a1-antitrypsin about 50% and for haptoglobin about 80% higher as compared to the results determined with the kinetic nephelometric method. An acceptable correspondence between the two methods was found for the results of transferrin.     

Prediction of myocardial infarction in dyslipidemic men by elevated levels of immunoglobulin classes A, E, and G, but not M

BACKGROUND: Immune mechanisms have been suggested to play an important role in the development of coronary atherosclerosis and its thrombotic complications. We evaluated the predictive value of the levels of various serum immunoglobulin classes in middle-aged men at increased risk of myocardial infarction. METHODS: Using nested case-control design and logistic regression analysis, we estimated the association between serum immunoglobulins and the risk of coronary end points (nonfatal or fatal myocardial infarction or sudden cardiac death) in dyslipidemic men (levels of non-high-density lipoprotein cholesterol >5.2 mmol/L [>201 mg/dL]) participating in the Helsinki Heart Study. The cases consisted of 135 subjects in whom a coronary end point occurred during the 5-year observation period of the study, and the controls were 135 subjects who did not suffer coronary end points during this period. Levels of IgA, IgE, IgG, and IgM were determined in serum samples collected at study entry. RESULTS: Levels of IgA, IgE, and IgG, but not IgM, were significantly higher in cases than in controls. After adjustment for other risk factors, such as age, smoking, and blood pressure, the risk of coronary disease showed a significant relation to the levels of IgA, IgE, and IgG. The risk in the highest quartile of each distribution as compared with the lowest quartile was 2.2-fold for IgA (95% confidence interval, 1.0-4.5), 2.8-fold for IgE (1.3-5.9), and 2.8-fold for IgG (1.3-5.9). Hypertriglyceridemia and a low level of high-density lipoprotein cholesterol were associated with increased risk of a coronary end point only if the levels of IgA, IgE, or IgG were also elevated. CONCLUSION: Elevated levels of IgA, IgE, and IgG are associated with myocardial infarction and cardiac death in men with dyslipidemia. The present data suggest that, for dyslipidemia to cause coronary atherothrombosis, an immune response reflected by elevated levels of these immunoglobulin classes is an important determinant.     

Local synthesis of immunoglobulins in neuropathies

It is essential to know how the immune system acts in different neurological diseases, some of them non very well known or of unknown etiology at all. It was applied Reiber and Felgenhauer's formula in 56 patients with different diseases. IgA, IgM, IgG and albumin were quantified in sera and cerebrospinal fluid by simple immunodiffusion. It was observed more frequent IgG local synthesis and IgA in this sample.     

Effect of fetal thymectomy on IgG, IgM, and IgA concentrations in sheep

Serum concentrations of immunoglobulins (Ig) IgG, IgG, IgM, and IgA were compared for normal and thymectomized lambs. Fetal thymectomies were performed in utero from 55 to 67 days of gestation. High serum IgG, IgM, and IgA concentrations occurred in all lambs after they ingested colostrum; however, the concentration of these Ig, as measured by single radial immunodiffusion, decreased exponentially during the first 16 days after birth. The half-life values for IgG, IgM, and IgA during this period in both normal and thymectomized lambs were about 25, 6, and 2 days, respectively. Increasing amounts of IgG were not detected in the serums of either group until 1 month of age. At 64 to 128 days, significantly smaller quantities of IgG and IgG were found in thymectomized lambs, whereas concentrations of IgA and IgM were similar in both groups. The results indicate that the thymus may regulate production of IgG in sheep.     

Detection of serum antibodies against Chlamydia pneumoniae by ELISA

Chlamydia pneumoniae causes pneumonia and other respiratory infections in children, adolescents and adults. We tried to evaluate the diagnostic value of detection of serum antibodies by ELISA for C. pneumoniae infections in Japanese children. Serum IgG, IgA and IgM antibodies to C. pneumoniae were determined by the microimmunofluorescence (MIF) test. Serum IgG and IgA antibodies were also determined by ELISA test kits. Results obtained by ELISA were compared with those obtained by MIF test. IgG antibody to C. pneumoniae was detected in 135 (39.5%) by ELISA and in 125 (36.5%) by MIF out of 342 sera from Japanese infants and children without respiratory infections (aged from 2 months old to 15 years old). IgA antibody to C. pneumoniae was detected in 129 (37.7%) by ELISA and in 117 (34.2%) by MIF out of 342 sera tested. Of 342 specimens 113 were IgG-positive by ELISA and MIF (sensitivity: 90.4%, specificity: 89.9%, r = 0.853). Of 342 sera 28 had IgG antibody titers of 1:256 and none had titers 1:512 or higher by MIF. Of 28 infants and children a total of nine were less than 4 years of age. On the other hand, of 342 specimens 99 were IgA-positive by ELISA and MIF (sensitivity: 84.6%, specificity: 86.7%, r = 0.769). Of 342 sera 16 had IgA antibody titers of 1:256 or higher by MIF. Of 16 infants and children, ten were less than 4 years of age. ELISA had excellent sensitivity and specificity relative to MIF test for detection of IgC and IgA antibodies to C. pneumoniae. It was suggested that C. pneumoniae infection in Japanese infants and children under 4 years of age was not infrequent.     

Comparison of cardiac findings at necropsy in octogenarians, nonagenarians, and centenarians

The Toxoplasma gondii rhoptry protein Rop2 was expressed in Escherichia coli as a fusion protein containing 44 kDa of the 55-kDa mature Rop2, supplied with six histidyl residues at the N-terminal end (Rop2196-561). Humoral response during Toxoplasma infection of humans was analyzed by immunoglobulin G (IgG), IgA, and IgM enzyme-linked immunosorbent assay with Rop2196-561 as the antigen substrate. The analyzed sera were divided according to T. gondii-specific serological tests (IgG, IgA, or IgM indirect immunofluorescence and IgA or IgM immunosorbent agglutination assay) as group A (IgG+ IgA- IgM-; n = 35), group B (IgG+ IgA+ IgM+; n = 21), group C (IgG+ IgA+ IgM-; n = 5), and group D (IgG+ IgA- IgM+; n = 16). Twenty-six T. gondii-seronegative sera from individuals with other infections were also included (group E). Anti-Rop2 IgG antibodies were detected in 82.8% of group A sera and in 97.6% of the sera with acute-phase marker immunoglobulins (groups B, C, and D). The percentage of IgA antibody reactivity against Rop2196-561 was 17.1% in group A, 50% in group D, and 80.8% in groups B and C. The percentage of IgM antibody reactivity was 0% in groups A and C and 62% in groups B and D. Sera from group E failed to show IgA, IgM, or IgG antibody reactivity. Since T. gondii Rop2 elicits a strong humoral response from an early stage of infection, it is suggested that recombinant Rop2196-561 would be suitable for use in diagnostic systems, in combination with other T. gondii antigens, to detect specific IgG, IgA, and IgM antibodies.     

Study of neonatal immunological function in breast feeding and formula feeding

OBJECTIVE: To determine 4 immunoglobulins and soluble interleukin-2 receptor (sIL-2R) in maternal and neonatal serum, in order to study the immunological functions in the neonates. METHODS: 80 cases were divided into three groups: (1) breast feeding group (30 cases), (2) formula feeding group (20 cases), and (3) mixed feeding group (30 cases). Maternal serum was collected prior to delivery and on the sixth day after delivery. Neonatal serum was collected on the third and sixth day after delivery. Umbilical blood at birth was obtained also. Secretory immunoglobulin A (SIgA), IgA, IgG, IgM and sIL-2R levels in the sera were determined by radioimmunoassay and enzyme-linked immunosorbent assay. RESULTS: The SIgA, IgA, IgG, IgM and sIL-2R levels in maternal serum were not significantly different among 3 groups, while neonatal serum SIgA, IgA, IgG, IgM and sIL-2R levels on the sixth day after delivery in the breast feeding group were significantly higher than those in the formula breeding group (P < 0.001, P < 0.001, P < 0.05, P < 0.05). CONCLUSION: Breast feeding may improve neonatal humoral and cellular immunity.     

Salivary immunoglobulin concentrations in patients with epilepsy treated with carbamazepine

Various anti-epileptic drugs may affect the immune system. An IgA-depressing effect of carbamazepine has been proposed, but only serum concentrations have been studied. IgA constitutes a small fraction of the serum immunoglobulins, while it is the predominating one in external secretions. In the present study the concentrations of IgA, IgG and IgM in unstimulated saliva were determined by single radial immunodiffusion in 34 patients with partial epilepsy, and being treated with carbamazepine alone. Median salivary IgA concentration in the patients was 208 x 10(-3) g/l, compared to 150 x 10(3) g/l in 41 healthy controls. Salivary IgG and IgM concentrations were also somewhat higher in the patients than in the controls, while the albumin concentrations were similar in the two groups. However, the differences in the immunoglobulin concentrations between patients and controls were not significant at a 5% level. There was no significant correlation between the concentrations of IgA in saliva and serum.     

Bilateral diffuse hypoperfusion of posterior temporoparietals and occipitals in Tc-99m HMPAO brain SPECT in a patient with noncommunicating hydrocephalus

We measured class-specific antibodies to the mycobacterial hsp70 protein in 67 patients with diabetes mellitus (27 type 1 and 40 type 2) with or without vascular complications. Using ELISA, the levels of IgG and IgM antibodies in the sera of diabetic patients did not significantly differ either from those of healthy control subjects or between both types of diabetes, regardless of gender, disease duration, HbA1 level, or type of vascular complication. In patients with type 2 diabetes, the mean serum IgA levels were significantly higher than those in their matched controls [274(71) mg/dl vs 208(88) mg/dl; P < 0.01]. In this group of patients, the IgA antibody titer was significantly correlated to the serum IgA level (r = 0.334; P < 0.01). Serological autoimmunity (IgG or IgM type) to hsp70 protein is common in both the normal and the diabetic population. The increased IgA levels and anti-hsp70 IgA titers in the sera of diabetics suggest a possible role of IgA in the pathogenesis of the vascular complications of diabetes mellitus.     

Effects of aluminum sulphate and citric acid ingestion on lipid peroxidation and on activities of superoxide dismutase and catalase in cerebral hemisphere and liver of developing young chicks

IgA antibodies reflecting airways or intestinal mucosal immune responses can be found in saliva and feces, respectively, and IgG antibodies reflecting serum antibodies can be found in saliva. In this study, antibodies were detected in samples of saliva and feces which had been air-dried at room temperature (+20 degrees C) or +37 degrees C, and stored at these temperatures for up to 6 months. In saliva the antibody levels increased, while the antibodies in feces decreased upon storage. The individual IgA antibody concentrations which were adjusted by using the ratios of specific IgA/total IgA were relatively stable in both saliva and feces, and correlated with corresponding antibody levels in samples which had been stored at -20 degrees C. The results indicate that air-dried saliva and feces can be used for semiquantitative measurements of mucosal antibodies, even after prolonged storage at high temperatures and lack of refrigeration.     

Kinetics of specific immunoglobulin A, M and G production in the duodenal and caecal mucosa of chickens infected with Eimeria acervulina or Eimeria tenella

The development and appearance of antibody was studied in the intestine and serum from histocompatible GB1 chickens orally infected with oocysts of Eimeria acervulina (restricted to the duodenum) or Eimeria tenella (restricted to the caeca). The local immune response was measured as the specific antibody levels in the supernatants of intestinal fragments (duodenum and caecum) maintained in culture for 16 h at 41 degrees C, 5% CO2, 95% air. Specific IgM was detected 1 week after E. acervulina infection, and the specific IgA and IgG contents of the duodenum and caecum were significantly elevated (P < 0.001) after 2 weeks. The intestinal specific IgG content was raised. E. tenella infection resulted in specific IgA only in the parasitized area during the second week post-infection (P < 0.05). Specific IgM and IgG were both detected in the duodenum and caecum, respectively, 1 and 2 weeks p.i. Production of parasite-specific immunoglobulins was always significantly higher in the parasitized than in the unparasitized areas (caeca for E. acervulina, duodenum for E. tenella). This ex vivo culture assay of intestinal fragments used to measure the mucosal immune response of intestinal areas showed a significant production of specific IgA and IgM. In addition, high levels of IgG were also measured. The role of this specific IgG in Eimeria infection remains to be determined.     

Immunoglobulin class and subclass distribution of antibodies reactive with the immunodominant antigen of Actinobacillus actinomycetemcomitans serotype b

The aims of this study were to determine the immunodominant antigens of Actinobacillus actinomycetemcomitans serotype b (Aab) for the different immunoglobulin (Ig) classes and subclasses and to determine the relative levels of these different Igs in serum. Seropositive early-onset periodontitis patients were sampled, and the Ig classes IgG, IgA, and IgM and subclasses IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2 were studied. Reactivity with Aab antigens was assessed by using the Western blot (immunoblot) in limiting dilution analysis and radioimmunoassay with sera from 13 early-onset periodontitis subjects. A smeared antigen in the upper portion of the immunoblots, typical of high-molecular-weight LPS, was immunodominant for IgG, IgA, IgM, IgG1, IgG2, IgG3, IgA1, and IgA2. This smeared antigen was present in every patient for all of these Igs at the endpoint. A few additional antigens were also present at the endpoint in some patients, but none were present in more than half of the subjects. The distribution of antibody titers by Ig classes reactive with the Aab immunodominant antigen was IgG > IgA > IgM. The distribution of antibody titers by IgG subclass was IgG2 > IgG1 approximately IgG3. Further quantitation by radioimmunoassay revealed that the mean concentration of IgG2 (65.7 micrograms/ml) was significantly greater than that of IgG1 (8.8 micrograms/ml). The IgA subclass distribution was IgA1 > IgA2, with IgA1 apparently being second only to IgG2. Therefore, the Aab antigen eliciting the highest antibody level in virtually all Ig classes and subclasses appeared to be lipopolysaccharide, and IgG2 was markedly elevated over all other serum Ig classes or subclasses reactive with Aab.     

Fetal immune response following prematurely ruptured membranes

Concentrations of immunoglobulins (Ig)A1, and IgA2, IgD, IgE, IgG, and IgM have been determined in cord blood, amniotic fluid, and maternal serum in a group of patients with a history of prematurely ruptured membranes (PRM) prior to the onset of labor and in a control group of patients undergoing normal delivery and without a history of infection during pregnancy. IgA and IgD were determined by sensitive hemagglutination-inhibition tests; IgG and IgM, by radial immunodiffusion; IgE, by a radioimmunoassay. There was evidence for an immune response in 10 of 16 cases of PRM: five of 16 had increased IgA but normal IgM; three of 16 had increased IgA and IgM; two of 16 had high IgM and normal IgA in cord blood. In patients with significantly increased levels of either IgA or IgM or both, there was a decreased level of IgD. These changes are most likely the result of the immune response to ascending infection from the maternal genitals. The sensitive testing method employed could demonstrate the presence of IgD in 53 per cent of normal cord blood samples and 72 per cent of amniotic fluid samples obtained at term. IgE was found in all normal cord blood and amniotic fluid samples tested. By concentrating the amniotic fluid up to 180-fold, IgM was demonstrated in all normal samples tested. The potential importance of IgA determinations in cord blood in addition to IgM determination for detection of intrauterine infections is stressed.     

Overexpression of c-myc induces apoptosis at the prophase of meiosis of rat primary spermatocytes

Serum samples from 36 cervical carcinoma patients, 33 patients with high-grade squamous intraepithelial lesions, and 31 cytologically normal women were tested by enzyme-linked immunosorbent assay (ELISA) using human papilloma virus type 6 (HPV 6) and HPV 16 virus-like particles as antigens. Forty serum specimens from 1-year-old children were used to assign cutoff points. When serum samples from the subjects infected with HPV 16 were tested in an HPV 16 ELISA detecting immunoglobulin A (IgA), IgG, and IgM binding, 61% showed IgA, 44% showed IgG, and 39% showed IgM reactivity. Of HPV 6- or 11- or HPV 18-infected subjects. fewer than 17% showed IgA or IgG responses and 33% showed IgM reactivity. In contrast, 13% showed IgA, 10% showed IgG, and 16% showed IgM reactivity in the HPV DNA-negative controls. The results suggest that the IgA and IgG responses are HPV 16 specific and the IgM response is cross-reactive to different HPV types. On the other hand, the serological responses to HPV 6 did not differ in the patient and control groups. The percentages of patients positive for both IgA and IgG antibodies were significantly higher in the groups with high-grade squamous intraepithelial lesions (12% [4 of 33]; P = 0.04) and cancer (17% [6 of 36]; P = 0.02) than in the healty women (0% [0 of 31]), and the percentages for either IgA or IgG were higher for the cancer group (47% [17 of 36]; P = 0.01) than in the normal group (19% [6 of 31]). Most sera positive for IgA and IgG in the patient groups showed higher titers than those in the normal group. All these results suggest that high IgA and IgG responses are good indicators for estimating HPV 16 infection.     

Collecting duct carcinomas of the kidney: a comparative loss of heterozygosity study with clear cell renal cell carcinoma

Lyophilizing was compared to freezing as a method of colostrum storage. Eight lots of colostrum from the first milking were divided into two equal parts; one was frozen, and the other was lyophilized. Twenty-two newborn calves were divided into two groups and fed either 2 L of frozen and thawed colostrum or 2 L of reconstituted lyophilized colostrum. The calves were bled at 12, 18, 24, and 72 h after feeding, and levels of the immunoglobulins IgG1, IgG2, IgM, and IgA were determined with a radial immunodiffusion assay, in colostrum and sera. The mean concentration of individual immunoglobulin isotypes in the sera of calves fed either frozen or lyophilized colostrum did not differ significantly. Calves fed from the same lots of colostrum had similar immunoglobulin concentrations in their sera, irrespective of the method of storage. All immunoglobulin isotypes were absorbed with equal efficiency from frozen and lyophilized colostrum as determined by calculation of the absorption coefficient.     

Experimental murine IgA nephropathy following passive administration of vomitoxin-induced IgA monoclonal antibodies

Oral exposure of mice to vomitoxin (VT) induces elevated levels of serum IgA, circulating IgA immune complexes (IgA-IC), mesangial IgA deposition and haematuria, which all mimic the clinical signs of human IgA nephropathy (IgAN). To further assess the effects of VT-induced IgA in the murine model, B6C3F1 and BALB/C mice were injected intraperitoneally with affinity-purified monoclonal IgA derived from Peyer's patch hybridomas of VT-exposed mice. In B6C3F1 mice, serum IgA, IgM and IgA-IC levels were increased two- to fivefold in treatment groups after 4 and 6 wk compared with controls, whereas increases in serum IgG as high as 18-fold were observed. Urinary erythrocyte counts were also significantly elevated in treatment groups after 2, 4 and 6 wk compared with controls. Concurrent increases in IgA and IgG complexes containing casein, the dietary protein source, occurred in treatment mice. Mesangial IgA, IgG, IgM and C3 deposition were significantly increased in all treatment mice after 6 wk. Electron-dense deposits occurred in the glomeruli of IgA-injected mice after 6 wk. All the above parameters were similarly affected in BALB/C mice. Injection of IgA-secreting hybridoma cells into BALB/C mice increased serum IgA, IgA-IC and IgG levels as well as elevated mesangial IgA, IgG and C3 deposition and haematuria after 2-3 weeks compared with controls. In total, these data indicate that passive administration of VT-induced IgAs can induce the hallmarks of IgA nephropathy. Casein, an antigen found in the diet used for these mice, appeared to form IC with IgA or IgG and these IC may participate in the pathogenesis of this nephropathy.     

Serum immunoglobulin levels in splenectomized Hodgkin patients and in subjects following post-traumatic splenectomy

Mean serum immunoglobulin levels (IgG, IgA, IgM) in splenectomized Hodgkin's disease patients in remission were compared with a control group of splenectomized healthy subjects following trauma. The controls showed a higher IgA and lower IgM than the normal. The IgM of the Hodgkin's group were decreased below the normal but showed no difference from the control group. IgA and IgG levels were not significantly different from the levels in the splenectomized healthy subjects. These results suggest that changes in immunoglobulins in splenectomized Hodgkin patients could be attributed to the effect of the removal of the spleen.     

Energetics of vinca alkaloid interactions with tubulin isotypes: implications for drug efficacy and toxicity

Capture ELISAs, for canine IgG, IgM, IgA and albumin, were developed and used to analyse immunoglobulin (Ig) concentrations in both serum and secretions. Matched samples of serum, saliva and tears were taken from 31 dogs, assigned to two groups based on age, whilst bile samples were obtained from nine adult dogs at post-mortem. Serum and tear IgA concentrations were significantly lower in dogs < or = 12 months of age compared with dogs > 12 months of age (p = 0.006 and 0.045, respectively). There was no significant correlation between serum and secretory Ig levels, with the single exception of serum and tear IgM concentrations (rp = 0.553, p = 0.004). IgG and IgM concentrations were significantly correlated in matched tear and saliva samples (IgG: rp = 0.470, p = 0.023; IgM: rp = 0.651, p < 0.0001). Albumin concentrations were significantly correlated with IgG, but not IgM or IgA, in both saliva and tears (saliva, rp = 0.581, p = 0.004; tears, rp = 0.843, p < 0.0001) whilst IgA and IgM concentrations were significantly correlated with each other in both secretions (saliva, rp = 0.644, p = 0.001; tears, rp = 0.555, p = 0.009). Significantly, more Ig of all classes was secreted into saliva than tears as calculated by a secretory index. A large diurnal and day-to-day variation was observed in Ig concentrations in serial saliva and tear samples taken from a further four dogs. Serum Ig concentrations are therefore, poor indicators of mucosal secretion in this species and significant intra-individual variation exists in secretory Ig levels. Both findings should be taken into account in future studies of canine mucosal immunoglobulins.