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PubMed1996

Evaluation of monoclonal antibodies with specificity for human IgA, IgA subclasses and allotypes and secretory component. Results of an IUIS/WHO collaborative study.

51 monoclonal antibodies (McAb) with putative specificity for human IgA, the IgA subclasses, Am allotypes or secretory component (SC) were evaluated for immunoreactivity and specificity by nine laboratories employing immunodiffusion, agglutination, immunohistological assays, immunoblotting and direct binding and competitive inhibition enzyme immunoassays. McAbs specific for IgA PAN (n = 24), IgA1 (n = 7), IgA2 (n = 3), IgA2m(2) (n = 2), non-IgA2m(2) (n = 4) and SC or secretory IgA (n = 5) were identified that were immunoreactive and specific in the assays employed. The McAbs identified as IgA- or SC-reactive were shown to be non-reactive to human IgG, IgM, IgD, IgE, kappa and lambda by direct binding and competitive inhibition immunoassays. Interestingly, no McAbs with restricted specificity for IgA2m(1) were identified. Some McAbs displayed higher affinity and/or better performance in one or several of the assay groups. The IgA- and SC-specific McAbs identified in this international collaborative study have potential as immunochemical reference reagents to identify and quantitate monomeric and polymeric IgA in human serum and secretions.

Animals↗
PubMed1988

Sandwich-type ELISA for free and bound secretory component in human biological fluids.

Three sandwich-type enzyme-linked immunosorbent assays (ELISA) are described for the measurement of free secretory component (SC) and SC bound to IgA (S-IgA) or IgM (S-IgM). These assays do not require preliminary fractionation of the biological fluids to be tested. The specificity of the assays is achieved with monoclonal antibodies specific for free SC (855 SC) and for SC bound to IgA or IgM (8545 SA). The amount of the three SC molecules in various biological fluids is reported. We demonstrate the presence of low levels of free SC in most of these fluids, including normal serum. Moreover our results suggest that S-IgM in serum may result from a non-covalent association between serum IgM and free SC.

Adult↗
PubMed1985

Regulation of secretory component by glucocorticoids in primary cultures of rat hepatocytes.

The present study examined the effects of steroid hormones on the production of secretory component (SC) by rat hepatocytes in cell culture. When hepatocytes were incubated in the presence of cortisol (10(-6) M), the levels of SC in media increased significantly after 2 days of incubation. This response was dose-dependent and specific for glucocorticoids because progesterone, dihydrotestosterone, and estradiol had no effect. When estradiol was added to the incubation media along with dexamethasone, a known potent synthetic glucocorticoid, it diminished the glucocorticoid response. The addition of cycloheximide to incubation media significantly decreased the effect of dexamethasone on SC accumulation. These findings suggest that glucocorticoid regulation of hepatocyte SC most likely involves stimulation of its synthesis. In addition, our results suggest that endogenous glucocorticoids may play a role in enhancing the clearance of IgA from blood into bile in the intact animal.

Animals↗
PubMed1991

Expression of HLA-DR and secretory component antigens and lymphocyte infiltration in human gastric nonmalignant and malignant tissues: an immunohistochemical study.

The relation between HLA-DR and secretory component (SC) expression and the degree of lymphocyte infiltration was immunohistochemically examined in human gastric mucosa with or without intestinal metaplasia and gastric carcinoma tissues. Gastric mucosa without obvious inflammation showed neither expression of HLA-DR or SC nor remarkable lymphoid infiltration. In contrast, gastric mucosa with chronic inflammation, tissues with incomplete type of intestinal metaplasia, and carcinoma demonstrated both HLA-DR and SC in almost the same area and also prominent lymphoid infiltration in the surrounding stroma. This simultaneous expression of HLA-DR and SC was not observed in complete type of intestinal metaplasia. The results indicate a close relationship between expression of HLA-DR and SC and the presence of lymphocyte infiltration in gastric mucosa, areas of incomplete type of intestinal metaplasia and in gastric carcinomas.

Adenocarcinoma↗
PubMed1976

Studies on translocation of immunoglobulins across intestinal epithelium. II. Immunoelectron-microscopic localization of immunoglobulins and secretory component in human intestinal mucosa.

To define mechanisms involved in the transport of immunoglobulins into intestinal fluids, we localized IgM, IgA, IgG, and secretory component (SC) in human intestinal mucosa by the peroxidase-labeled antibody technique. At the light microscopic level, immunocytes containing IgA, IgM, or IgG were found in the lamina propria. IgA, IgM, and SC were prominent in the epithelium of gland crypts; IgG was limited to a few cells at tips of villi. At the electron-microscopic level, SC was localized to perinuclear spaces, endoplasmic reticulum, saccules associated with Golgi complexes, cytoplasmic vesicles, and lateral and basal plasma membranes of columnar epithelial cells. IgA and IgM, but not IgG, also were localized to plasma membranes and cytoplasmic vesicles of these cells. Neither the immunoglobulins nor SC was found within other types of epithelial cells (Paneth, goblet, endocrine). The findings provide evidence that (1) the site of SC synthesis in intestinal epithelium is secretory columnar cells, principally those in gland crypts; (2) the polymeric immunoglobulins IgM and IgA are translocated through such SC-containing cells by a process that involves formation of cytoplasmic vesicles; (3) IgM and IgA could combine with SC during transcellular transport (likely sites are lateral or basal plasma membranes or supranuclear cytoplasm); (4) the monomeric immunoglobulin IgG does not share the transepithelial cell route involved in IgM and IgA transport.

Biological Transport↗
PubMed1987

Early detection of liver metastasis in patients with colorectal carcinoma by increased levels of circulating IgA- and IgM-associated secretory component.

One hundred patients operated for colorectal carcinoma were followed clinically and with serial blood samples from 5 to 8 years. Levels of secretory component (SC) associated with IgA and IgM in serum were measured and related to Dukes' stage, histological differentiation, tumour expression of SC, and circulating carcinoembryonic antigen (CEA). On the whole, elevated levels of SC in serum were found in 15 of the 20 patients who already had (n = 15), or later developed (n = 5), liver metastasis. Four of the latter 5 patients showed raised SC levels with a 5.5 months median lead time from the first positive serum sample to clinically manifest liver disease. These data are interesting in view of the promising results reported for liver resection in patients with colorectal carcinoma.

Adult↗
PubMed1984

Histological and histochemical studies of the secretory components of the salamander olfactory mucosa: effects of isoproterenol and olfactory nerve section.

Secretory components of the salamander olfactory mucosa, sustentacular cells (SC), and Bowman's glands (BG), were examined histologically and histochemically. In the aquatic larval salamander, SC in sensory grooves contained secretory granules; the submucosa contained a single layer of homogeneous, ductless glands. In the land-dwelling adult salamander, SC spanning a flat epithelial sheet contained vesicles. Subjacent to the epithelium in both dorsal and ventral mucosae lay BG whose ducts opened at the surface of the epithelium. In the ventral mucosa, two additional layers of olfactory glands (OG) lying below the BG were identified; ducts were not observed in association with the OG. The beta-adrenergic agonist isoproterenol caused depletion of secretory granules from BG and OG of larval, young, and adult salamanders but had no discernible effect on SC. Histochemical techniques (Alcian blue at pH 2.5 and pH 1.0, high-iron diamine, and the periodic acid-Schiff reaction) demonstrated that SC contained neutral, acidic, and small amounts of sulfated mucopolysaccharides (MPS), BG and OG contained only neutral MPS. In contrast, glands under adjacent respiratory epithelium contained both acidic and sulfated MPS. Unilateral olfactory nerve section ( ONX ) caused changes in the histochemical reactivity of acidic and sulfated MPS in SC on the ipsilateral and later on the contralateral side. Neutral MPS staining became enhanced first in the OG that lay under the BG, then in BG cells, and later in the deepest OG layer. Ipsilateral changes preceded contralateral ones. At 24 days post- ONX , some acinar cells in the deep OG contained acidic but not sulfated MPS.

Animals↗
PubMed1993

Secretory component and lactoferrin in pure pancreatic juice in chronic pancreatitis.

To evaluate pathophysiological roles of proteins in pancreatic secretion, immunoreactive lactoferrin (LF) and secretory component (SC) were measured in the first fraction of the pure pancreatic juice obtained endoscopically from 17 control, 21 suspected (SCP), 14 noncalcified (NCP), and 14 calcified chronic pancreatitis (CCP) subjects. The protein and amylase tended to decrease both in concentration and output from control to CCP. LF concentration was elevated in CCP (18.0 +/- 4.9 micrograms/ml) when compared with controls (2.3 +/- 0.2 micrograms/ml), and LF output in NCP (12.3 +/- 3.8 micrograms/min) was increased from controls (3.8 +/- 0.6 micrograms/min). The combination of high LF concentration with low protein output was observed in 10/14 in CCP but 0/14 in NCP and can be a biochemical discriminator of CCP from NCP. SC concentrations were also elevated in NCP (8.5 +/- 2.0 micrograms/ml) and CCP (5.6 +/- 1.6 micrograms/ml) from controls (1.2 +/- 0.2 micrograms/ml). SC outputs in SCP (9.8 +/- 3.1 micrograms/min) and NCP (21.1 +/- 4.8 micrograms/min) were increased from controls (1.7 +/- 0.3 micrograms/min), but there was no further increase in CCP. Hypersecretion of LF and SC in chronic pancreatitis is different, especially in CCP, although the mechanisms for hypersecretion are unknown.

Adult↗
PubMed1993

Surface expression of secretory component and HLA class II DR antigen on glandular epithelial cells from human endometrium and two endometrial adenocarcinoma cell lines.

The expression of secretory component (SC) by human glandular endometrial cells cultured in vitro was significantly increased by estradiol in the medium. Interferon-gamma and interleukin-4 stimulated the expression of SC only in the presence of estrogen. Tumor necrosis factor-alpha plus estrogen also caused a significant increase in the number of cells expressing SC. HLA class II antigen DR was detected on few glandular epithelial cells of human endometrium cultured in control medium, whereas interferon-gamma and interleukin-4, but not tumor necrosis factor-alpha, caused significant increases in the expression of DR. Estrogen in the culture medium did not significantly affect DR expression. The human endometrial adenocarcinoma cell lines, HEC and RL-95, expressed SC in approximately 50 and 20% of the cells. Also, approximately 20% of the RL-95 cells stained for DR antigen. Interferon-gamma did not influence the degree of expression of either surface marker of the two cell lines. Cells of both lines bound polymeric IgA and IgM but showed little to no binding of monomeric IgA, IgG, or an IgM previously shown not to bind SC.

Adenocarcinoma↗
PubMed1999

Production of secretory component and pathogenesis of gastric cancer in Helicobacter pylori-infected stomach.

The purpose of this study was to examine the production of secretory component (SC) and immunoglobulin A (IgA) in the gastric mucosa with Helicobacter pylori infection and to investigate the influence of immunological reactions on various phases of infection (gastritis, intestinal metaplasia, gastric cancer). Production of SC and IgA was assessed by immunohistochemical staining in (1) endoscopic biopsy samples of H. pylori-eradicated cases (n = 25), and (2) surgically resected stomach tissues of H. pylori-positive gastric cancer cases, intestinal type (IGC, n = 25) and diffuse type (DGC, n = 25). Before eradication therapy, all samples showed positive staining of SC and IgA in epithelial cells, and IgA was also positive in plasma cells in the mucosal layer. H. pylori bacteria were positively stained for SC and IgA. After treatment, the degree of SC and IgA staining in epithelial cells was reduced with successful eradication; but with intestinal metaplasia, SC staining was positive regardless of the results of treatment. In nonmetaplastic mucosa, SC-positive cells were increased in the glandular neck zone to the surface mucosal layer; and the intensity of SC staining was increased in proportion to the degree of mucosal inflammation and IgA-positive cell aggregation. In intestinal metaplasia, SC was positive irrespective of the degree of inflammation. Most cancer foci also showed positive staining of SC, irrespective of histological type. Production of SC and IgA was thought to be a specific reaction against H. pylori infection, occurring from the early to the late stages and not limited to intestinal metaplasia. It was suggested that immunological reactions against H. pylori infection might generally be involved with the pathogenesis of intestinal metaplasia and both histological types of gastric cancer (IGC and DGC).

Gastric Mucosa↗
PubMedDate not supplied

Secretory component in human amniotic fluid and gestational tissues: a potential endogenous phospholipase A2 inhibitor.

OBJECTIVES: Prostaglandins (PGs) are essential mediators of labor during human pregnancy. Phospholipase A2 (PLA2) provides the essential substrate for PG synthesis through the liberation of arachidonic acid from membrane phospholipid stores. Nonlaboring amniotic fluid (NL-AF) contains secretory component (SC)-like protein(s) that suppress in vitro PLA2 activity. This study characterizes the biologic activity, identity, and tissue distribution of these protein(s) in NL-AF and gestational tissues. METHODS: Third-trimester NL-AF was collected by amniocentesis, fractionated by ammonium sulfate precipitation, and submitted to an in vitro PLA2 assay. Identity of the PLA2 inhibitor in NL-AF was confirmed by Western blot and antibody neutralization studies. Secretory component-immunoreactive proteins were purified by immunoaffinity chromatography and visualized by sodium dodecyl sulfate-gel electrophoresis. Tissue distribution of SC in gestational tissues was determined by immunohistochemistry. RESULTS: The 100% pellet and supernatant fractions of NL-AF suppressed PLA2 activity, and this activity was neutralized by a polyclonal antibody to SC. Western blot studies revealed an SC-reactive protein in the 70-80-kD range in the 100% pellet fraction of NL-AF. Two SC-reactive proteins were detected in the 60-80-kD range in the eluate from the SC immunoaffinity column, along with minor proteins of 30 and greater than 100 kD. Immunohistochemical studies revealed SC in placental trophoblast, amniotic membranes, and decidual epithelium. CONCLUSIONS: These results demonstrate that proteins homologous to SC are present in human gestational tissues and possess anti-PLA2 activity. These proteins may contribute to the maintenance of pregnancy by suppressing local PG production.

Amniocentesis↗
PubMed1984

Immunohistochemistry of carcinoembryonic antigen, secretory component and lysozyme in benign and malignant common bile duct tissues.

An immunoperoxidase technique has been utilized for the localization of carcinoembryonic antigen (CEA), secretory component (SC) and lysozyme ( LZ ) in normal and cancerous common bile duct tissues. Little or no CEA was found in the non-cancerous common bile duct tissues. SC was found in the surface epithelium and accessory gland epithelium and LZ was demonstrated only in the accessory glands. Some inflammatory cells were also positively stained for LZ . In adenocarcinoma, CEA was always present on the luminar border of the carcinoma cells, occasionally with intercellular and intracellular localization. LZ was absent, or only faintly detected in carcinoma. SC was generally distributed in well-differentiated adenocarcinoma cells, but showed a reduced intensity of staining with progressive dedifferentiation. These findings suggest that CEA, SC and LZ could be useful markers providing valuable information in the pathological diagnosis of bile duct carcinoma.

Adenocarcinoma↗
PubMed1984

Estradiol regulation of secretory component in the uterus of the rat: evidence for involvement of RNA synthesis.

The present studies were undertaken to characterize the response of uterine secretory component (SC) to estradiol. Administration of estradiol for 3 days to ovariectomized rats before incubation of uterine tissues resulted in a marked accumulation of SC in the incubation media. When uteri from ovariectomized rats treated with progesterone or testosterone were incubated, very little SC accumulated in the media, indicating that the estradiol-stimulated increase is hormone-specific. When uteri from rats that received estradiol for 6 days were compared with uteri from 3-day treated rats, SC release during a 24-hr incubation period was the same. This finding indicates that in the presence of prolonged estradiol exposure, SC production continues. The estradiol-induced accumulation of SC in culture is not due to the release of pre-formed uterine SC. When tissue SC levels were measured after 3 days of estradiol treatment, very little tissue SC was found relative to that released into culture media during 24 hr of incubation. The addition of actinomycin D to the incubation media markedly inhibited SC release by uteri from estradiol-treated rats. The release of SC was also inhibited by alpha-amanitin, a known inhibitor of Type II polymerase. These studies demonstrate that estradiol stimulation of SC is markedly reduced by inhibitors of RNA synthesis, and suggest that estradiol regulation of SC is mediated through uterine mRNA synthesis.

Animals↗
PubMed1986

Human secretory component. IV: Antigenic regions involved in in vitro binding to dimeric IgA.

The aim of this report was to identify the region(s) of the secretory component (SC) molecule involved in in vitro binding to dimeric IgA. Inhibition of the SC binding was tested by Fab' antibody fragments directed against the accessible (A) and inaccessible (I) regions of SC. Antibodies directed against the main 38.5-kDa trypsin fragment of SC, and antibodies from two immune sera with a wide anti-SC spectrum, were also used. The specificity and activity of the five antibody preparations were established by double-diffusion in gel and by their SIgA combining capacity. Inhibition curves were established by RIA using constant amounts of 125I SC, dimeric IgA and increasing quantities of the various Fab' antibodies. These results indicated involvement of a larger part than the I region of the SC molecule in combination with dimeric IgA, perhaps including a second (minor?) site of binding on the accessible parts in addition to the major region located on I.

Antibody Specificity↗
PubMed1988

Lateral diffusion of the secretory component (SC) in the basolateral membrane of the human colon carcinoma cell line HT29 assessed with fluorescence recovery after photobleaching.

The lateral diffusion of the secretory component (SC), acting as a receptor for dimeric IgA in the basolateral side of intestinal epithelial cells, was studied in the human colonic carcinoma cell line HT29. The HT29 cells were grown in Dulbecco's modified Eagle's medium in which galactose had been substituted for glucose to promote development of small intestine-like cells, with a distinct separation of the basolateral side from the apical surface. The SC was stained with rhodamine-labeled polyclonal anti-human SC rabbit antibodies (Ig) or Fab fragments, and the lateral mobility was assessed with the fluorescence recovery after photobleaching technique. The average lateral diffusion was consistent with a diffusion constant of 7.7 +/- 2.0 (mean value +/- SD; n = 29) and 7.1 +/- 2.3 (n = 30) x 10(-10) cm2s-1 for Ig-and Fab-labeled receptors, respectively, which is slower than lipid diffusion but is similar to that found for other membrane receptors. The corresponding values for the fraction of mobile receptors were 66 +/- 13% and 71 +/- 12%, respectively. Cells were labeled from the top of the culture plate, and cells adjacent to a mechanically made rift or a natural opening in the cell monolayer were labeled more strongly, confirming the microscope-based impression that the basolateral surface primarily harboured the SC receptor.

Colonic Neoplasms↗
PubMed1984

Evaluation of nine different fixatives. 1. Preservation of immunoglobulin isotypes, J chain, and secretory component in human tissues.

Eight cross-linking fixatives were tested for their ability to preserve IgG, IgA, IgM and IgD isotypes, J chain, and secretory component (SC) in paraffin-embedded specimens of human tonsils and colonic mucosa. The results were compared with the antigenic preservation afforded by cold 96% ethanol (with or without inclusion of a prefixation 48-hr washing period). A semiquantitative immunofluorescence scoring system was applied. In relation to the cytoplasmic scores of 3.0 assigned to IgG and IgA immunocytes after ethanol fixation, the following median scores were obtained with the other fixatives: routine formalin, 1.3 (IgG and IgA); glutaraldehyde (1%)-formalin, 0.3 (IgG and IgA); Baker's formol calcium, 2.3 (IgG) and 1.5 (IgA); formol sublimate, 1.0 (IgG) and 1.2 (IgA); acetic acid (2%)-formol saline, 2.0 (IgG and IgA); Bouin's fluid and Susa fixative, 0 (IgG) and 1.3 (IgA); and carbodiimide, 2.3 (IgG) and 2.0 (IgA). Most aldehyde-based fixatives afforded poorer result for IgA immunocytes when an anti-alpha-chain reagent of restricted specificity was applied. The result was usually slightly better for IgM cells than for IgG cells, but IgM did not resist fixation in Bouin's fluid and Susa fixative to the same extent as IgA. IgD immunocytes were poorly revealed except after fixation with Baker's formol calcium, carbodiimide, and acetic acid-formol saline, which afforded median fluorescence scores of 1.0-1.8 compared with 3.0 after ethanol fixation. J chain in IgA immunocytes was most intensely stained in sections of ethanol-fixed tissue denatured in acid urea (median score, 3) but was also well revealed after fixation with formalin, acetic acid-formol saline, Bouin's fluid or Susa fixative (median scores, 1.8-2.0). The results with most of the cross-linking fixatives were much less favorable for IgA, IgM, J chain and SC in colonic crypt epithelium than in immunocytes. Only carbodiimide afforded fluorescence scores for IgA and SC approaching those obtained after ethanol fixation (2.0 and 1.5 vs. 3.0 and 2.8). The effect of proteolytic unmasking depended on the fixative - not only for different antigens and locations in the tissue, but also for the same antigen in apparently similar cells.

Colon↗
PubMed1983

Immunohistochemical study of the distribution of secretory component and IgA in the normal and diseased uterine mucosa.

The distribution of secretory component (SC) and IgA in the normal and diseased uterine mucosa was studied by means of an immunoperoxidase method. Both substances could not be demonstrated in ectocervical epithelium, whereas in endocervical epithelium both were present and staining intensity appeared to be related to the degree of inflammatory activity in the underlying stroma. Local IgA-containing plasma cells most likely are the source of epithelial IgA. Well-differentiated adenocarcinomas of the endocervix were shown to display SC immunoreactivity. In endometrial glands SC was observed in the postovulatory phase of the menstrual cycle, in increasing quantities toward menstruation. Hyperplastic conditions of the endometrium showed a weak focal staining for SC. In contrast, in most well-differentiated adenocarcinomas of the endometrium, SC and IgA could be detected in most neoplastic elements, whereas poorly differentiated carcinomas were largely negative. It is concluded that: (a) in contrast to the endometrium, in the endocervix a local secretory immune system operates similar to that of the gastrointestinal tract; (b) the ability of the endometrium to synthesize SC is at least partly regulated by hormonal factors and, as such, is confined to the progestagen-dominated phase of the menstrual cycle; and (c) immunohistochemical detection of SC may be of help in the grading of adenocarcinomas of the endometrium.

Adenocarcinoma↗
PubMed1989

Immunohistochemical localization of secretory component and immunoglobulin A in the urogenital tract of the male rodent.

The mucosal immune system in the male rodent urogenital tract was studied by localizing secretory component (sc) in the rat and immunoglobulin A (IgA) in both rat and mouse by immunofluorescence. In the rat, bright labelling of sc was observed at several sites, including the ejaculatory ducts, excretory ducts of several accessory glands, and urethral glands in the pelvic and bulbous portions of the urethra. Pale labelling of sc was detected in epithelial cells of the ventral prostate gland. Plasma cells containing IgA were only observed in the urethral gland in the bulbous portion of the urethra in rats and mice. These results suggest that IgA may be transported into the urogenital tract of the male rat primarily at sites distal to the production of seminal fluid and spermatozoa. While locally synthesized IgA may be available in the bulbous urethra, it appears that serum may be the main source of IgA for transport into the rat urogenital tract at the other sites where its receptor, sc, was demonstrated.

Animals↗