Characterization of circulating and cutaneous IgA immune complexes in patients with dermatitis herpetiformis

Dermatitis herpetiformis (DH) is a chronic, blistering skin disease characterized in part by deposits of IgA at the dermal-epidermal junction. Eighty-five percent of DH patients have granular IgA deposits and have an associated gluten-sensitive enteropathy (GSE). In contrast, 15% of DH patients have a linear pattern of IgA deposits and no associated intestinal abnormality. Although circulating IgA antibodies against skin are not present in these patients, 40% of DH patients do have IgA-containing circulating immune complexes (IgA-CIC). The role and origin of the cutaneous IgA and the IgA-CIC in patients with DH are unknown; however, the association of GSE with the granular IgA deposits suggests that a mucosal immune response may be important in the pathogenesis of DH. We have characterized the IgA subclass composition of the cutaneous IgA deposits in patients with DH, and have isolated and characterized the IgA-CIC from these patients. Twenty-nine of 29 patients with DH and granular IgA deposits were found to have only IgA1 deposits. Ten of 11 patients with linear IgA deposits also had only IgA1 deposits; one of 11 had IgA2 deposits. Isolated IgA-CIC from the sera of eight patients with DH and granular IgA deposits were found to contain both IgA1 (58% +/- 5, mean percent of total IgA +/- SEM) and IgA2 (42% +/- 5), as were IgA-CIC from two patients with ordinary GSE without cutaneous IgA deposits. The IgA subclass composition of the isolated immune complexes was significantly different from the serum IgA1 and IgA2 composition (serum IgA1 = 76% +/- 6; IgA2 = 24% +/- 5, p less than 0.025, Student's t-test), and suggests that the IgA-CIC may arise from gut-associated lymphoid tissue (GALT). Sequential anti-IgA1 absorption of serum which contained IgA-CIC did not remove all the IgA-CIC, suggesting that the complexes circulate as separate IgA1 and IgA2 complexes. The finding of IgA1 alone in the skin of patients with DH suggests that the cutaneous IgA may not arise from GALT, or that IgA1, possibly arising in GALT, is preferentially bound to DH skin. Because IgA-containing CIC which contain both IgA1 and IgA2 were found in the serum of patients with DH and with ordinary GSE, it seems unlikely that IgA-containing CIC are responsible for the cutaneous IgA deposits seen in DH.     

Immunological follow-up of hydatid cyst cases

Hydatid disease is caused by Echinococcus granulosus. In this study, we aimed to investigate the benefit of monitoring cases with hydatid cyst by means of immune components in patients in a long-term follow-up after surgery. Eighty-four preoperative and postoperative serum samples from 14 cases undergoing surgery for hydatid disease were evaluated in terms of immune parameters, such as total and specific IgE, IgG, IgM, IgA and complement. Total and specific IgE were determined by ELISA. Specific IgG levels were measured by indirect hemagglutination.Total IgG, IgM, IgA and complement (C3 and C4) were detected by nephelometry. Imaging studies were also carried out during the follow-up. In none of the patients hydatid cysts were detected during the follow-up. Total IgE levels in the sera of the patients decreased to normal six months after surgery. Although specific IgE against echinococcal antigens decreased one year after operation, levels were still significantly high. There were no changes in the levels of anti-Echinococcus IgG and total IgG in follow-up period. Additionally, other parameters, such as IgA, IgM, C3 and C4, were not affected.     

Serum and secretory immunoglobulins in allergic diseases

A total of 158 patients with pollinosis, bronchial asthma, urticaria and Quincke's edema were examined. The immunoglobulin and C3 levels in sera and the immunoglobulin and albumin levels in saliva were determined by the method of single radial immunodiffusion with the corresponding monospecific antisera. In all the groups of patients subjected to examination the presence of polyclonal hypergammaglobulinemia was detected, which was manifested by a rise in the levels of IgG, IgA and especially IgM; the level of IgD was low. A decrease in the level of C3 was detected in pollinosis patients in the absence of the exacerbation of the disease. No circulating immune complexes were detected. An essential increase in the level of IgG in saliva was revealed, which was due to the local synthesis of this immunoglobulin. In winter the level of salivary IgA in pollinosis patients was found to be essentially below normal, but at the period of exacerbation it increased twofold, probably in response to local stimulation with antigen-allergen. Patients with bronchial asthma and pollinosis were found to have a high level of free secretory component (SC); in pollinosis the level of free SC sharply increased during the stage of exacerbation, which was due to the increase of its synthesis and secretion by the epithelial cells of the mucous membranes. The importance of these data for the pathogenesis of allergic diseases are discussed.     

Prevalence of Helicobacter pylori infection in Warao lineage communities of Delta Amacuro State, Venezuela

The purpose of this study was the evaluation of Helicobacter pylori infections in children and adults from two indigenous communities of Delta Amacuro State, Venezuela, that differ in hygienic conditions of the housing. The evaluation was performed in 98 children (mean age 7 +/- 3.37 years) and their mothers (33.96 +/- 13.77 years) from two communities of Warao lineage. Anti-H. pylori serum IgG and secretory anti-H. pylori IgA antibodies were determined, as well as total secretory IgA and H. pylori antigens in feces. Serological prevalence of H. pylori infection was 38% in children and 84% their in mothers. Children from the community that had the most deficient sanitary and hygienic conditions had significantly lower titers of specific IgG antibodies and total secretory IgA (P<0.0001) and a high percentage of them had H. pylori antigens in their feces (P<0.0001). The levels of specific IgA were similar in both groups. The results indicate that in these populations there is a high prevalence of H. pylori infection and that poor hygienic conditions can increase the risk of infection and damage to the gastrointestinal tract.     

Humoral response of the immunologic secretory system of the small intestine in children infected with Giardia intestinalis

Local humoral response of the intestinal mucosa was determined with secretory IgA levels and secretory component activity in enterocytes and duodenal content of 15 children infected with G. intestinalis. The obtained results were compared to those in 5 children with coeliac disease and 12 children with diarrhoea without lambliasis. Secretory IgA was increased in about 50% of children with lambliasis (in the remaining groups in 25% of children) to the values higher than that in the comparative groups. Secretory component activity was relatively high in the intestinal epithelium. Secretory component activity in the duodenal content was high in about 40% of children independently of the examined group. No correlation between the said variables was noted except positive correlation of secretory IgA levels and secretory component activity in the bile.     

Characterization of anti-Cryptosporidium IgA antibodies in sera from immunocompetent individuals and HIV-infected patients.

Human antibody response to Cryptosporidium parvum has been previously shown as involving immunoglobulin (Ig)M and IgG isotypes. The interest in anti-cryptosporidial IgA antibody response has been recently stimulated by studies on the therapeutic effects of secretory IgA antibodies to Cryptosporidium in animal models and in patients. In the present study, isotypes of serum anti-Cryptosporidium antibodies have been characterized in donors of the following categories: (a) healthy adults, (b) healthy children, (c) immunocompetent children with transient cryptosporidial diarrhea, (d) HIV-infected patients without clinical and parasitological evidence of Cryptosporidium infection and (e) AIDS patients with cryptosporidial diarrhea. Antibodies were detected using C. parvum oocysts purified by density gradient centrifugation from bovine faeces. The IgA antibodies were revealed using alpha-chain specific antibodies. Indirect immunofluorescence analysis with oocysts was used as control. Although high levels of serum antibodies of the IgA class were detected in some donors in the group of healthy adults, elevated values were consistently found in HIV-infected patients. Higher values were found in HIV patients with clinical cryptosporidiosis. The presence of a secretory component in serum IgA antibodies in these patients has been documented. Data indicate that IgA serum antibodies are produced as well as IgM and IgG antibodies upon contact with the parasite, and suggest that elevated IgA serum antibodies to Cryptosporidium are not associated with protection in HIV patients.     

Humoral immunity in malnutrition

Malnutrition affects the humoral immune system in diverse fashions. B lymphocyte subpopulations, serum IgG, and IgA levels, and immunoglobulin synthesis and metabolism are usually normal or increased. Hypogammaglobulinemia may occur in very young, severely malnourished infant. Although usually normal, deficient antibody responses to injected antigens are occasionally present, depending on the severity of the malnutrition. Serum IgE levels are usually high in malnutrition, probably due both to the increased incidence of intestinal parasites and to the lack of T cell control of IgE. Despite these high IgE levels, allergy is unusual. Secretory IgA levels in the respiratory and gastrointestinal (GI) fluids are generally decreased, as are secretory IgA antibody responses. This local antibody deficiency may increase GI permeability, stimulate immune complex formation, and result in the formation of IgG food antibodies. Breast milk secretory IgA levels are equivalent in malnourished mothers but because of lessened milk volume, the total amount of secretory IgA delivered to their offspring is diminished. Complement levels are somewhat diminished in the presence of malnutrition with a resultant opsonic deficiency. Although these B cell system aberrations undoubtedly contribute to the enhanced susceptibility to infection of malnourished patients, they are usually less severe than are concomitant T cell deficiencies.     

抗小鼠精子特有乳酸脱氢酶C4的与分泌片装配或未装配的单克隆抗体IgA的生物学功能

为了测定抗精子ＩｇＡ在免疫不育和抗精子避孕疫苗研制方面的生物学作用,用肠道内免疫的方法制备了一组抗乳酸脱氢酶Ｃ４（ＬＤＨ－Ｃ４）的单克隆ＩｇＡ抗体（ｍｏＩｇＡ）。以免疫印迹证实了它们的异质同形体。大部分ｍｏＩｇＡ（ＰＡ１－ＰＡ５）是用肠道内免疫和以派依尔氏淋巴细胞作为亲本细胞进行融合来获得的。在豚鼠血清补体存在的情况下,小鼠精子可以被ｍｏＩｇＡＰＡ１、ＰＡ２和ＰＡ４所制动。高浓度ＰＡ４和ＰＡ５可凝集小鼠精子。小鼠体外受精率可被３个ｍｏＩｇＡ（ＰＡ２、ＰＡ３和ＰＡ４）显著降低,但用ＰＡ１、ＰＡ２和ＰＡ５被动免疫之后,小鼠体内受精无明显变化。纯化的小鼠胆汁分泌片可同纯化的ｍｏＩｇＡ或腹水中的ｍｏＩｇＡ在体外组装起来。同分泌片结合之后,ｍｏＩｇＡ对精子的制动、凝集和体外受精无明显变化。这些研究结果提供了抗ＬＤＨ－Ｃ４的ｍｏＩｇＡ和分泌性ＩｇＡ对精子功能和体外受精的生物学作用的直接证据,在免疫不育的防治,避孕疫苗的研制以及性传播疾病的防治方面均有一定的指导意义。     

Mucosal B cell deficiency in IgA-/- mice abrogates the development of allergic lung inflammation

We have investigated the consequence of lack of IgA on host immunity using a murine model of allergic lung inflammation. Mice with a targeted disruption of the alpha-switch region and 5' H chain gene (IgA(-/-) mice), which lack total IgA, developed significantly reduced pulmonary inflammation with fewer inflammatory cells in lung tissue and bronchoalveolar lavage fluids, as well as reduced levels of total and IgG1 OVA-specific Abs and decreased IL-4 and IL-5 in bronchoalveolar lavage fluids compared with IgA(+/+) controls, following allergen sensitization and challenge. This defect was attributable to fewer B cells in the lungs of IgA(-/-) mice. Polymeric IgR-deficient (pIgR(-/-)) mice, which lack the receptor that transports polymeric IgA across the mucosal epithelium where it is cleaved to form secretory IgA, were used to assess the contribution of secretory IgA vs total IgA in the induction of allergic lung inflammation. pIgR(-/-) and pIgR(+/+) mice had comparable levels of inflammation, demonstrating that IgA bound to secretory component is not necessary for the development of allergic lung inflammation, although this does not necessarily rule out a role for transudated IgA in lung secretions because of "mucosal leakiness" in these mice. The results indicate that Ag-specific B cells are required at mucosal surfaces for induction of inflammation and likely function as major APCs in the lung for soluble protein Ags.     

Secretory IgA and secretory component in women affected by recidivant vaginal candidiasis

Local immunity was evaluated in 47 patients affected by recidivant vaginal candidiasis and 33 control women. IgG, IgA, IgM and secretory component (SC) were determined by single radial immunodiffusion in samples of cervicovaginal secretion. IgG in dosable levels was detected in 17/47 samples (36.2%) and IgA in 15/47 patients (31.9%) whereas in the controls, the incidence was 31/33 (93.9%) for IgG and 24/33 (72.7%) for IgA. The difference was significative (P< 0.001) for both immunoglobulins. Significant differences were not obtained for IgM. The SC was detected in 4/47 cervicovaginal secretions of patients affected by candidiasis (8.5%) whereas in the control samples the incidence was 21/33 (63.6%) (P<0.001). In only 2/15 patients with dosable levels of IgA (13%) the secretory nature of this immunoglobulin could be shown by its reaction with anti-SC serum. In the control group, secretory IgA was detected in 19/24 cases (79%) (P< 0.001). Serum immunoglobulins levels were normal. The lack of secretory IgA and SC in the secretion could be related to the adherence capacity of the Candida albicans to epithelial cells.     

Polymeric IgA rheumatoid factor in idiopathic IgA mesangial nephropathy (Berger's disease)

A specific and sensitive enzyme-linked immunosorbent assay (ELISA) was used to detect IgA rheumatoid factor (RF) in sera from 88 patients with IgA nephropathy (IgA GN), a disease characterized by abnormalities of IgA production. Significantly higher levels of IgA antiglobulins were demonstrated in IgA GN patients than in normal healthy controls and patients with other forms of chronic primary glomerulonephritis (mean +/- SEM 28.4 +/- 6.6 vs 6.0 +/- 0.4 and 8.3 +/- 1.2 micrograms/ml respectively; p less than 0.002). Interestingly, in contrast to rheumatoid arthritis, IgA RF activity was not associated with IgM antiglobulins. Analysis of sera fractionated by gel chromatography at acid pH revealed that anti-IgG activity resided predominantly in the polymeric fractions of IgA as confirmed by the ability to bind "free" secretory component. Several findings in patients with IgA GN suggest that the IgA deposited in the glomeruli is polymeric, and levels of circulating macromolecular IgA are increased. Our findings confirm a general perturbation of IgA metabolism in this disease. Although the polymeric nature of the IgA RF is suggestive of a mucosal origin, additional evidence is needed to confirm this hypothesis.     

Heterophile Hanganutziu-Deicher antibodies in sera of patients with Kawasaki diseases

The heterophile antibody levels in sera from patients with Kawasaki disease (49 sera from 39 cases) were measured by sheep erythrocytes (SRBC) agglutination and radioimmunoassay in microplates coated with Hanganutziu-Deicher (H-D) antigen-active glycosphingolipid, equine hematoside. The antibody levels were low in the first week of illness, increased rapidly in the 2nd week, and thereafter gradually decreased. The SRBC agglutination titers and H-D antibody titers of sera from patients with Kawasaki disease from week 2 to 8 of illness were significantly higher than those of healthy children (44 sera) and normal cord blood (13 sera).     

Quantification of human urinary free secretory component, secretory and nonsecretory IgA by ELISA

The specific quantification of human urinary free secretory component (FSC), secretory IgA (SIgA) and total IgA using ELISA has been hampered by mutual interferences of these three molecules. Using affinity chromatographically purified antisera an attempt was therefore made to reduce these interferences without necessitating further assay steps. FSC and total IgA were measured in unprocessed urine by means of anti-FSC and anti-IgA as well as alkaline phosphatase-coupled anti-FSC or anti-IgA antisera. SIgA was determined using anti-IgA as well as alkaline phosphatase-coupled anti-FSC. Nonsecretory urinary IgA was calculated from the measured SIgA and total IgA. The mutual interferences of FSC, SIgA or nonsecretory IgA in the three assay systems were low and not relevant for normal samples. Normal urinary concentrations were: FSC 344 +/- (SD) 208 ng/ml (n = 120), SIgA 1,874 +/- 1,133 ng/ml (n = 123) and nonsecretory IgA, depending on the way of standardization, 712 +/- 699 (n = 56) or 878 +/- 732 ng/ml (n = 51). SIgA excretion increased with age. Lower urinary SIgA as well as total and nonsecretory IgA levels were observed in males as compared to females. No correlation evolved between the hormonal status of women and the excretion of FSC, SIgA or IgA. In IgA-deficient patients virtually no nonsecretory IgA or SIgA was detected in the urine while the FSC concentration was in the normal range.     

类风湿关节炎患者RF、ANA、CCP、Ig及炎症因子的水平测定及临床意义

目的:探讨类风湿关节炎(RA)患者血清类风湿性因子(RF)、抗核抗体(ANA)、抗环瓜氨酸肽(CCP)抗体、免疫球蛋白(Ig)、补体(C3、C4)以及炎症因子的水平及临床意义。方法:收集2016年9月至2017年4月我院收治的165例RA患者为RA组,其中RA活动期患者93例(RA活动组),RA缓解期患者72例(RA缓解组),并于同期随机选取30例健康体检者为对照组。采用免疫散射比浊法检测各组血清RF、IgM、IgG、IgA、C3、C4水平,采用酶联免疫吸附法(ELISA)检测各组血清CCP抗体,电化学发光法检测白介素-6(IL-6)、化学发光法检测白介素-8(IL-8)。免疫荧光法检测ANA。比较不同组别各检测指标水平,并分析RA患者RF、ANA、CCP抗体、Ig、C3、C4与炎症因子的相关性。结果:RA活动组、RA缓解组血清RF、ANA、CCP抗体、IgM、IgG、IgA、IL-6、IL-8水平高于对照组,且RA活动组血清RF、ANA、CCP抗体、IgM、IgG、IgA、IL-6、IL-8水平高于RA缓解组,差异均有统计学意义(P0.05)。RA活动组、RA缓解组血清C3、C4水平低于对照组,且RA活动组血清C3、C4水平低于RA缓解组,差异均有统计学意义(P0.05)。经Pearson积矩相关分析,RA活动期和缓解期患者血清RF、ANA、CCP抗体、IgM、IgG、IgA与炎症因子IL-6、IL-8呈正相关关系(P0.05),血清C3、C4与炎症因子IL-6、IL-8呈负相关关系(P0.05)。结论:RA患者体内RF、ANA、CCP抗体、Ig及IL-6、IL-8水平明显较高,C3,C4水平明显较低,活动期RA患者更为显著,联合检测可早期辅助诊断RA及判断病情进展,在临床上有重要的参考意义。     

Development and validation of a sensitive ELISA for quantification of secretory IgA in rat saliva and faeces.

Non-invasive measures of immunological markers are an attractive means of stress assessment in laboratory animals. Salivary IgA has been used successfully as a stress marker in the human, and several reports indicate the potential of secretory IgA as a non-invasive measure of stress in animals. The present paper describes the development of an ELISA using commercially available components for the quantification of rat IgA and validation of this assay for the quantification of rat secretory IgA in saliva and faeces. The concentration of IgA in rat saliva varied significantly between duplicate samples obtained from individual rats, and the viscosity and small total volume of rat saliva gave unsatisfactory results for IgA. Faecal IgA was present in high concentrations, and duplicate samples varied by only 2-3%. However, faecal IgA seemed less stable than IgA in other biological compartments, and this finding must be taken into consideration when using quantitative measurements of IgA as a marker of mucous humoral immune status.     

丹参对毛细支气管炎患儿体液免疫调节作用的观察

目的:探讨丹参佐治毛细支气管炎的疗效及其可能的机理。方法:73例毛细支气管患儿随机分为常规组和丹参组,均用常规方法治疗,但后者加用复方丹参静脉滴注。在治疗前、后用全自动散射比浊法测定并比较两组患儿血清IgE、IgG、IgA、补体C3和C4水平。结果:丹参组和常规组总有效率依次为94.44%和78.38%,差异具有统计学意义(P<0.05)。治疗后,丹参组患者血清IgE、C3和C4水平下降,IgG、IgA水平上升,与治疗前相比均有一定程度差异(P<0.01-0.05)。治疗后,常规组患者仅IgE水平下降(P<0.01),但其余指标与治疗前差异无统计学意义(P>0.05)。结论:丹参佐治毛细支气管炎,临床效果较好,且可调节机体的体液免疫应答,增强机体的抗感染能力。     

Platelet immune complex interaction in pathogenesis of Kawasaki disease and childhood polyarteritis.

The role of platelets in the pathogenesis of vasculitis and the formation of coronary artery aneurysms was studied in 19 children with Kawasaki disease and five with polyarteritis. All patients with Kawasaki disease developed thrombocytosis in the third week of illness. The peak platelet count was significantly correlated (p less than 0.005) with the subsequent development of coronary artery aneurysms. The rise in platelet count was associated with the appearance in the circulation of a factor that induced aggregation and serotonin release in normal platelets. This factor was shown to be of high molecular weight, and its activity was lost at low pH--features suggestive of an immune complex. Immune complexes, detected by precipitation with polyethylene glycol, also appeared in the circulation as the platelet count increased. These complexes induced platelet aggregation, and there was a significant correlation (p less than 0.001) between the concentrations of IgG and IgA in the polyethylene glycol precipitated material and the platelet aggregating activity. Similar platelet aggregating activity was also detected in patients with polyarteritis but followed a different time course, persisting in the circulation for several months in association with continued disease activity. These findings imply that different mechanisms have a role in distinct phases of Kawasaki disease. The initial feverish phase (probably infective) is probably followed by an immune complex vasculitis that occurs when antibodies to the initiating agent appear in the circulation. The immune complexes aggregate platelets and induce release of serotonin. Platelet derived vasoactive mediators may increase vascular permeability and facilitate further deposition of complexes in the tissues.     

Isolated glomerulonephritis with mesangial IgA deposits.

Mesangial deposits of IgA, occurring in the absence of systemic disease known to be associated with nephritis, were detected by immunofluorescence microscopy in renal biopsy specimens from 25 patients (4% of 630 specimens studied). Associated deposits of C3 were always present, usually with IgG, but IgM deposits were less common and C1q was never seen. On light microscopy most of the biopsy specimens showed mesangial of focal nuclear proliferation though some were normal. Fifteen of the 25 patients presented with macroscopic haematuria, which was usually recurrent and preceded by a sore throat, whereas the remaining, and usually older, patients presented with persistent proteinuria and were more likely to have impaired renal function. This incidence of "mesangial IgA disease" is less than that reported by French workers. There was a significantly high incidence of familial renal disease among these patients. No abnormalities of serum complement or IgA concentration were found.     

IgA and IgG antigliadin,IgA anti-tissue transglutaminase and antiendomysial antibodies in patients with autoimmune thyroid diseases and their relationship to thyroidal replacement therapy

Celiac disease is a chronic illness of the small bowel caused by gliadin intolerance in genetically predisposed subjects. The aim of this study was to investigate serum levels of IgA and IgG antigliadin antibodies, IgA antiendomysial antibodies, and IgA anti-tissue transglutaminase antibodies in 169 patients with autoimmune thyroid diseases, i.e. chronic thyroiditis and Graves' disease. Antiendomysial antibodies were positive in 2 out of 169 persons (1.18%), IgA antigliadin antibodies in 15.98%, IgG antigliadin antibodies in 51.48%, and IgA anti-tissue transglutaminase in 14.79%. The prevalence of positivity was higher compared to the 1312 control blood donors described in our previous study (Vancíková et al. 2002) (p<0.05). Patients with chronic thyroiditis treated with a high replacement dosage of levothyroxin (125-200 microg daily) had higher serum levels of IgA antigliadin antibodies in comparison with patients treated with a lower dosage (50-100 microg daily) (medians: 13.00 vs. 19.69, p=0.033). We found a negative correlation of IgA anti-tissue transglutaminase antibodies and total calcium serum levels (r = -0.480, p=0.0236, n=22). We can conclude that in persons with autoimmune thyropathy there is a high prevalence of positive antigliadin, anti-tissue transglutaminase and antiendomysial antibodies. Latent celiac disease may lead to impaired resorption of therapeutically administered levothyroxine, calcium, or other substances.