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Effect of thermal injury in the rat on transfer of IgA protein into bile.

Severe thermal injury is associated with bacterial sepsis; the intestine is considered a likely source of invasive organisms. Because IgA antibody in bile accounts for much of the specific immune defense of the upper intestinal tract in the rat, the effect of thermal injury on the quantity of IgA protein in bile was examined. Sprague-Dawley rats received a 20% to 30% body surface area burn under anesthesia. Eighteen hours later the common bile duct was cannulated and bile was collected for three hours. Total IgA protein in bile decreased 90% after thermal injury. The bile volume, the concentration of bile protein, and free secretory component did not change significantly. Although blood flow to the liver 18 hours after thermal injury was not changed, there was a significant reduction in total IgA concentration in the circulation; both monomeric (m-IgA) and polymeric IgA (p-IgA) were decreased. This finding may explain, in part, the reduced concentration of IgA protein in bile. Although not examined in this study, decreased local hepatic synthesis and/or transport of p-IgA across the hepatocyte may also contribute to the reduced IgA levels in bile.

Animals↗

Possible mechanisms of elevation of serum secretory immunoglobulin A in liver diseases.

In order to investigate possible mechanisms of elevation of serum secretory immunoglobulin A (sIgA) in liver diseases, human liver specimens were applied to immunohistochemical study of immunoglobulin A, secretory component, and J chain, which are components of sIgA. In the cases of chronic hepatitis with high serum sIgA levels, these antigens were present in dilated bile canaliculi of hepatocytes and they were continuously stained on the lateral plasma membrane of hepatocytes from the bile canaliculus to the space of Disse over the junctional complexes. Furthermore, in liver cirrhosis and extrahepatic cholestasis, they were also detected in intraportal bile ductules and intercellular spaces of degenerated cholangiocytes. These results suggest that at least two pathways might allow elevation of serum sIgA: through the communication of the bile canaliculus with the space of Disse over junctional complexes and through the bile ductule into the portal blood vessel.

Bile Canaliculi↗

An enzyme-linked immunosorbent assay for differential quantitation of secretory immunoglobulins of the A and M isotypes in human serum.

An enzyme-linked immunosorbent assay was developed for differential quantitation of secretory IgA (SIgA) and secretory IgM (SIgM) in human serum. The assay was based on non-competitive binding of SIgA and SIgM to microplates coated with an excess of antibodies to secretory component (SC). Appropriate standards were included to obtain absolute values. Mutual competition of SIgA and SIgM was avoided by testing the serum samples at sufficiently high dilutions. The assay is fast, simple, sensitive and reproducible. All of the 138 healthy individuals tested (1-91 years old) were found to have both SIgA and SIgM in their serum (medians, 10 mg/l and 14 mg/l, respectively). Lactating women, SIgA-deficient healthy individuals, and particularly patients with hepatitis had significantly increased serum SIgM levels compared with controls. Differential quantitation of SIgA and SIgM may turn out to be of diagnostic value and provide pathogenetic information.

Adolescent↗

The pulpal origin of immunoglobulins in dentin beneath caries: an immunohistochemical study.

Immunoglobulins localized in uninfected dentin beneath caries are thought to be protective, but their origin remains controversial. We reasoned that the localization and dominance of serum IgG1 would support the pulpal origin of the immunoglobulins while a predominance of secretory component (SC) bearing IgA1 and IgA2 would support their salivary origin. The prevalence and staining intensity of IgG1, IgA1, IgA2, IgM, and SC in uninfected dentinal tubules beneath shallow, deep caries, and noncaries teeth were examined immunohistologically. SC was only localized in caries, and IgG1 was the predominant subclass in uninfected dentinal tubules beneath shallow and deep caries, followed by IgA1. In noncaries teeth, IgG1 was localized on the pulpal end. The intensity of IgG1 was significantly higher than either IgA1 or IgA2 in both shallow and deep caries. Our data support the serum origin of immunoglobulins in uninfected dentin beneath caries.

Analysis of Variance↗

Reduced secretion of IgA to skin surface of patients with atopic dermatitis.

To investigate whether the secretory form of immunoglobulin A (sIgA) was reduced on the skin surface in atopic dermatitis, the amount of sIgA present in sweat was measured in 40 patients with atopic dermatitis and in 50 healthy volunteers by attaching a cellulose membrane disk (10 x 10 mm) to the inner aspect of the upper arm skin for 24 hours. The secretory form of IgA, which was absorbed to the membrane and accumulated during the period of application, was revealed as dots by an enzyme immunoassay in which antibodies for IgA and for the secretory component were used. The density and number of dots (per mm2/day), which corresponded to the openings of eccrine excretory ducts, were determined with a densitometer. The mean amount of sIgA secreted by those patients was 3.86 +/- 0.71 pg/mm2/day (range, 0 to 21.17 pg/mm2/day), whereas that of the control subjects was significantly higher (p < 0.001), 16.79 +/- 2.80 pg/mm2/day (range, 0.79 to 133.77 pg/mm2/day). This may be related to the high incidence of bacterial and viral skin infections seen in patients with atopic dermatitis, and in addition, to the development of eczematous lesions through a defect in ridding the skin of allergens and/or microorganisms.

Adolescent↗

Antibody against the human J chain inhibits polymeric Ig receptor-mediated biliary and epithelial transport of human polymeric IgA.

To emphasize the requirement for a J chain in native polymeric immunoglobulins for their selective transport into exocrine secretions, IgG, purified from two different antisera specific for the human J chain, was shown to: (i) bind in vitro to human polymeric IgA (pIgA) by density gradient ultracentrifugation; (ii) inhibit binding in vitro of rat secretory component to human pIgA; (iii) inhibit hepatic transport of human pIgA into rat bile in vivo; and (iv) inhibit apical transcytosis of pIgA in vitro by polarized human polymeric immunoglobulin receptor (pIgR)-expressing Madin-Darby canine kidney cells. Inhibition of biliary transport increased with the molar ratio of anti-J chain antibodies against pIgA and their incubation time. Anti-J chain F(ab')2 and Fab fragments also inhibited biliary transport, excluding a role for phagocytic clearance or excessive size of the immune complexes. Anti-human-Fc alpha Fab, bound to human pIgA in complexes of larger size than those with anti-J chain Fab, did not inhibit biliary transport of human pIgA. Propionic acid-denatured human pIgA, although containing J chains, was very poorly transported into rat bile. Altogether, our data strongly support, now also by in vivo experiments, the crucial role of the J chain of native pIgA in its selective pIgR-mediated transport into secretions, as suggested long ago by in vitro data only. Recent data on J chain-knockout mice, with low IgA levels in bile and feces, cannot explain the role of the J chain in contributing to the secretory component/pIgR-binding site of normal pIgA, but otherwise agree with our study.

Animals↗

Regulation of Candida albicans growth and adhesion by saliva.

To examine the local regulation of oral Candida albicans growth, we examined non-stimulated and stimulated salivary flow rates (SFRs) and the C. albicans growth and adhesion inhibitory activities of saliva in 60 patients with oral candidiasis (divided into two groups: 25 patients with oral candidiasis only (group OC) and 35 patients with oral candidiasis and systemic diseases (group CS)) and 30 healthy control subjects. Both non-stimulated and stimulated SFRs in patients, especially in group CS; were decreased in comparison with those in the healthy control subjects. The levels of secretory immunoglobulin A (sIgA) in group OC and group CS and the lactoferrin level in group CS were decreased as compared with those in control individuals, although there were no differences in transferrin and total secretory component (SC) levels between the three groups. The secretion amounts (microg/min) of these proteins were statistically significantly decreased in the patients, especially in group CS. Saliva from the patients showed a lesser inhibitory effect on C. albicans growth and adhesion to HeLa cells than did saliva from the control subjects. In addition, polymorphonuclear leukocytes (PMNs) in patients' saliva generated smaller amounts of superoxide than did those in control subjects' saliva, and phagocytic and C. albicans killing activities were suppressed in the patients. These results indicate that the decreases in SFR, secretion of antimicrobial proteins in saliva, and salivary PMN activity are risk factors for oral candidiasis associated with aging and systemic diseases.

Adult↗

The DNA content of colorectal carcinomas: an analysis of the heterogeneity of aneuploidy and correlation with immunopathological parameters.

DNA ploidy patterns of 107 colorectal adenocarcinomas were retrospectively analyzed by flow cytometry (FCM) and correlated with clinicopathological and immunohistological parameters as grade, vascular and serosal invasion, carcinoembryonic antigen, secretory component (SC) and expression of HLA-DR antigens. Using the classical division into DNA diploid and DNA aneuploid tumors, a correlation with SC expression was observed. Subdivision of aneuploid tumors according to the discussed hypothetical functional criteria into hyperdiploid, hypotetraploid, tetraploid and hypertetraploid groups led to statistically significant correlation with serosal invasion. Among the immunohistological parameters the interdependence of SC and HLA-DR expression may be also of interest. The need for subdivision of DNA aneuploidy is broadly discussed so as to permit the analysis of different properties, cellular origins and kinetic parameters. Ultimately this may lead to more clinically meaningful interpretation of DNA histograms.

Adenocarcinoma↗

Origin of immunoglobulins in respiratory tract secretion and saliva of sheep.

The origin of the immunoglobulins in the upper respiratory tract secretion of sheep was determined by measuring the distribution between plasma and secretion of radiolabelled purified immunoglobulins and albumin. By calculation of the ratio of specific activity for each immunoglobulin between plasma and secretion, it was estimated that about 81% of IgA in secretion was of local origin, whereas IgM, IgG1, IgG2 and albumin were wholly derived from plasma. Estimates of the selectivity of transport of IgA and IgM into both respiratory tract secretion and saliva were obtained by calculation of a selective index relative to IgG1 or IgG2, which do not bind secretory component (SC). This was based on radioactivity ratios after the simultaneous injection of immunoglobulin labelled with different isotopes (IgA or IgM injected with either IgG1 or IgG2). These calculations revealed that both IgA and IgM were selectively transported into respiratory tract secretion and saliva. This provides further support for the proposition that SC-binding immunoglobulins may be transported from serum into secretions at a variety of mucosal sites dependent on SC availability. Since the IgA in serum of sheep is predominantly of gut origin, this provides an opportunity, in addition to relocation of gut-derived plasma cell precursors, by which the gut may contribute to extraintestinal mucosal responses.

Albumins↗

Detection of monomeric and polymeric IgA containing immune complexes in serum and kidney from patients with alcoholic liver disease.

The purpose of this study was to characterize circulating IgA and the IgA deposited in the glomeruli of patients with alcoholic liver disease. In the 6 patients studied there was an increased proportion in monomeric IgA (3.5 fold) and IgA between 9-13S (8.94 fold), 13-17S (4.49 fold) and 17-21S (1.63 fold) fractions on 5-40% sucrose density gradient ultracentrifugation at physiological pH. All fraction between 9.12S decreased at acid pH, however a 3.28 fold increase in fractions where polymeric IgA is expected to appear. IgG eluted at acid pH from autopsy kidney was studied by the same procedures. At pH 7.4 about 55% of that IgA have a molecular weight comprised between 9-12S, decreasing to around 25% at acid pH. The existence of true polymeric IgA in serum and kidney was based on the capacity of high molecular weight IgA to bind human secretory component. The amount of immune complexes with monomeric IgA were higher than those with polymeric IgA in serum as well as in kidney. However, the percentage of heavy IgA (probably polymeric IgA) in kidney were, in each patient, higher than those observed in serum. Our results show the presence of high amounts of monomeric and polymeric IgA, both partially as immune complexes, in serum and kidneys of patients with alcoholic liver disease and IgA glomerulonephritis. Furthermore, our data suggest a role for human liver in the clearance of serum IgA such as has been demonstrated in the some animal species, especially in rats.

Antigen-Antibody Complex↗

Oral tonsils: an immunoperoxidase study.

Tissue morphometry and the distribution of cells containing immunoglobulin (Ig), J chain and secretory component (SC) were studied in eight oral tonsils. Quantitative immunohistochemistry revealed that IgG-containing cells predominated in all lymphoid compartments (follicles, extrafollicular areas and reticular epithelium) and that the IgG:IgA:IgM class ratios of the overall tonsillar immunocyte population were 13:8:2. Cells containing IgD and IgE were rare. IgG immunocytes showed no significant localisation within a given lymphoid compartment, whereas IgA cells were found predominantly in extrafollicular areas, especially adjacent to surface epithelium, and IgM cells were in follicles. J chain was present within IgM and some IgA cells. Tonsillar crypt and surface epithelium was negative for SC, suggesting that these structures are not directly involved in local mucosal immunity.

Humans↗

Lectin receptors on IgA isotypes.

It has been shown previously that secretory IgA interacts with the mannose-specific lectin of Escherichia coli. The purpose of the study described here was to evaluate whether the N-linked oligosaccharide chains of the human IgA isotypes IgA1 and IgA2 differ in lectin receptor activity. A range of plant lectins specific for N-linked oligosaccharide chains were tested for their ability to precipitate IgA1 and IgA2 myeloma proteins, secretory IgA and free secretory component. IgA2 myeloma proteins reacted more strongly than IgA1 with the mannose-specific lectin ConA, whereas IgA1 myeloma proteins reacted more strongly than IgA2 with two galactose-specific lectins, Ricinus communis agglutinin I and Abrus precatorius agglutinin. This suggests that IgA2 possesses a larger proportion of short truncated complex type oligosaccharide chains and/or oligomannose type chains than IgA1. Further, IgA2 reacted more strongly than IgA1 myeloma proteins with Lens culinaris (lentil) lectin, and Pisum sativum (pea) lectin, suggesting that IgA2 exposes more of short, complex type chains fucosylated on the core than IgA1. The differences demonstrated in receptor activity between IgA1 and IgA2 may be important in their interaction with the microbial flora, as well with endogenous lectins, such as phagocyte receptors.

Carbohydrate Sequence↗

Transepithelial transport of immunoglobulins.

Immunoglobulins are transported across a variety of epithelial tissues. The best studied example of this is the transport of polymeric IgA and IgM by the polymeric immunoglobulin receptor (pIgR) across many types of epithelial cells. The pIgR binds its ligand at the basolateral surface and is internalized into endosomes. Here it is sorted into vesicles that transcytose it to the apical surface. At the apical surface the pIgR is proteolytically cleaved, and the large extracellular fragment (known as secretory component) is released together with the ligand. The pIgR contains a cytoplasmic domain of 103 amino acids that contains several sorting signals. Targeting from the trans-Golgi network to the basolateral surface is determined by the membrane-proximal 17 residues of this domain. There are two endocytosis signals that contain crucial tyrosine residues. Transcytosis of the pIgR is stimulated by binding of polymeric IgA. Phosphorylation of a cytoplasmic serine promotes transcytosis of the pIgR without ligand bound. Transcytosis may be regulated by the heterotrimeric Gs protein, protein kinase C and calmodulin. IgG is transcytosed from the apical to basolateral surface in several epithelial tissues such as the placenta and the small intestine of newborn rats. The receptor for intestinal transport of IgG is structurally similar to class I MHC molecules.

Animals↗

Interactions of human mesangial cells with IgA and IgA-containing immune complexes.

BACKGROUND: IgA nephropathy (IgAN) is characterized by IgA1-containing immune complexes in mesangial deposits and in the circulation. The circulating immune complexes (CIC) are composed of galactose- (Gal) deficient IgA1 and IgG or IgA1 antibodies specific for the Gal-deficient IgA1; interactions of these CIC with mesangial cells (MC) were studied. METHODS: Binding, internalization, and catabolic degradation of myeloma IgA1 protein as a standard control and the isolated CIC were studied using human MC, hepatoma cell line HepG2 expressing the asialoglycoprotein receptor (ASGP-R), and monocyte-like cell line U937 expressing the Fc(alpha)-R (CD89). Biochemical and molecular approaches were used to assess expression of CD89 and ASGP-R by MC. RESULTS: At 4 degrees C, radiolabeled IgA1 bound to MC and HepG2 cells in a dose-dependent and saturable manner. The binding was inhibited by IgA-containing CIC or excess IgA1 or its Fc fragment but not by the Fab fragment of IgA1. At 37 degrees C, the cell-bound IgA1 was internalized and catabolized. In addition to IgA1, HepG2 cells also bound (in a Ca2+-dependent manner), internalized, and catabolized asialoorosomucoid (ASOR), other asialo-(AS)-glycoproteins, and secretory component (SC). The binding by MC appeared to be restricted to IgA1 or AS-IgA1 and was not Ca2+-dependent. Furthermore, MC and HepG2 cells internalized and catabolized IgA1-containing CIC. Using RT-PCR with ASGP-R- or CD89-specific primers, mRNAs of the two respective genes were not detected in MC. CONCLUSIONS: The data showed that the ability of MC to bind IgA1 and IgA1-containing CIC in vitro was mediated by an IgA receptor that was different from CD89 or ASGP-R and had a higher affinity for IgA-CIC than for uncomplexed IgA.

Antibody Affinity↗

Elevated levels of secretory immunoglobulins A and M in serum of patients with large bowel carcinoma indicate liver metastasis.

Preoperative serum levels of secretory IgA (SIgA) and secretory IgM (SIgM) were quantified by an enzyme-linked immunosorbent assay in 100 patients with large bowel carcinoma. The values obtained were related to tumor characteristics such as Dukes' stage, differentiation, DNA ploidy pattern, expression of secretory component (SC), volume, and plasma level of carcinoembryonic antigen (CEA). Statistical comparison was made with matched controls. Only patients with Stage D tumors showed significantly increased serum levels of SIgA and SIgM (P less than 0.004). The combined diagnostic sensitivity of SIg and CEA in Stage D was 0.94. In patients with liver involvement, serum SIgA and SIgM were elevated in 67% and 53%, respectively. However, circulating SIg was neither correlated with tumor SC expression nor with any other studied variable. Thus, the raised serum SIg levels were apparently not caused by release of SC from the tumors but, instead, by hampered liver function due to hepatic metastasis.

Adult↗

Selective transport of IgA. Cellular and molecular aspects.

The principal characteristic immunoglobulin of mucosal surfaces, secretory immunoglobulin A (S-IgA), is the product of two different types of cell present in mucosal and glandular tissues. Submucosal plasma cells, which are generated largely within the common mucosal immune system, synthesize predominantly polymeric, J chain-containing IgA, which is selectively bound by polymeric immunoglobulin receptor or secretory component (SC) on the basolateral surfaces of mucosal and glandular epithelial cells. The molecular and cellular events involved in SC expression, its intravesicular transport together with its polymeric IgA ligand to the apical surface of the epithelial cell, during which IgA becomes covalently linked to SC, and the proteolytic cleavage of SC from the apical membrane to release S-IgA into the lumen have been elucidated. Additional receptors and mechanisms for the uptake, catabolism, and transport of IgA exist, especially in the liver. The biologic significance of IgA transport lies in the secretion of large quantities of S-IgA antibodies for the protection of huge areas of mucosal surfaces and for the provision of passive immunity to suckling infants, and in the immune elimination of antigenic materials by hepatobiliary transport.

Animals↗

Tryptic digestion of bovine secretory IgA at elevated temperature and in urea. Isolation of SC domain 1 which is covalently bound to IgA dimer and binds non-covalently to IgM.

1. Tryptic cleavage sites in bovine secretory component (SC) which become inaccessible when SC is bound to IgA dimer remained inaccessible at 60 degrees C and in 4 M urea at 37 degrees C. 2. This suggests the presence of strong interactions compatible with published affinity constants of ca 10(8) M-1. 3. In 5 M urea at 37 degrees C further cleavage of bound SC did occur to produce a fragment consisting of domain 1 which was disulphide bridged to the IgA dimer. 4. Binding studies on the isolated fragment showed that domain 1 did not account for all the binding by SC. 5. Cleavage of the isolated fragment with iodosobenzoic produced a smaller fragment consisting of the n-terminal third of domain 1 (residues 1-35). This N-terminal fragment showed significant binding.

Alkylation↗

Immunohistochemical study of pulmonary adenocarcinoma.

In order to improve the accuracy of diagnosis and subtyping of pulmonary adenocarcinomas, immunohistochemical studies were carried out on 105 adenocarcinomas of the lung procured from both surgery and autopsy. Avidin-biotin-peroxidase complex methods were used for identifying keratin, vimentin, carcinoembryonic antigen (CEA), and secretory component (SC) on deparaffinized tissue sections. Keratin was positive in 29% of well differentiated adenocarcinoma, significantly lower than in moderately or poorly differentiated adenocarcinoma. Likely, vimentin was positive in 27% of well differentiated adenocarcinoma, significantly lower than in moderately or poorly differentiated adenocarcinoma. SC was positive in 66% of well differentiated adenocarcinoma, significantly higher than in moderately or poorly differentiated adenocarcinoma. In the subtyping of well differentiated adenocarcinomas, keratin showed higher positive results in the bronchial surface epithelial, goblet cell, and bronchial gland types than in the Clara cell or type II alveolar epithelial cell type. These findings suggest that immunoperoxidase stains for keratin, vimentin, and SC may be useful for determining the degree of differentiation of adenocarcinomas of the lung as well as for subtyping of well differentiated pulmonary adenocarcinomas.

Adenocarcinoma↗