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PubMed1987

Rat mast cells permeabilised with streptolysin O secrete histamine in response to Ca2+ at concentrations buffered in the micromolar range.

Rat peritoneal mast cells have been permeabilised by treatment with streptolysin O which generates membrane lesions of macromolecular dimensions. In the presence of Ca2+ buffered at concentrations in the micromolar range, the permeabilised mast cells release histamine, beta-N-acetylglucosaminidase and lactate dehydrogenase. Release of the two secretory components (but not lactate dehydrogenase) has an obligatory requirement for a nucleoside triphosphate and micromolar concentrations of Ca2+. Inosine triphosphate (ITP) supports the release reaction better than ATP does. It is concluded that the secretory materials are released from the cells by an exocytotic mechanism, while lactate dehydrogenase leaks from the cells through the toxin-generated lesions. By initially withholding and then supplying Ca2+ to the permeabilised cells, it is shown that the exocytotic secretory reaction can persist even when the cytosol is depleted of the bulk of soluble proteins. The streptolysin O treated mast cell preparation represents a simplified system with which to study the mechanism of exocytosis.

Acetylglucosaminidase↗
PubMed1993

Malignant oncocytoma of the parotid gland: a case report with an immunohistochemical and ultrastructural study.

A rare case of a 71-year-old man with malignant oncocytoma is presented. The tumour was found in the left parotid region with enlarged lymph nodes in the neck. The resected parotid mass consisted of three discrete nodules, each histologically characterized by a uniform proliferation of oncocytic tumour cells. The patient underwent repeated operations for recurrences and metastases, and eventually died of acute pneumonia 18 months after the first admission. Metastases to the lymph nodes, ribs, spine and liver had been clinically pointed out. Immunohistochemically, the tumour cells were positive for alpha-1-antitrypsin, alpha-1-antichymotrypsin, lactoferrin, secretory component and carcinoembryonic antigen (CEA), while they were negative for S-100 protein and HHF35 (muscle specific actin). Ultrastructural pictures disclosed numerous mitochondria in the cytoplasm of the tumour cells, revealing neither myofibrils nor secretory granules. These findings therefore support the concept that this tumour is of a glandular epithelial origin.

Adenoma↗
PubMed1988

Polymeric and monomeric IgA response in serum and milk after parenteral cholera and oral typhoid vaccination.

The effect of vaccinating lactating Pakistani mothers with a combination of live oral typhoid vaccine and parenteral inactivated cholera vaccine on specific milk and serum IgA antibodies in both monomeric (m) and polymeric (p) forms was analysed. IgA antibody titres peaked for both antigenic specificities 2 weeks after the first dose of vaccine. 82 +/- 7% of anti-Vibrio cholerae and 72 +/- 17% of anti-Salmonella typhi IgA were in the polymeric form. These serum pIgA antibodies were mainly dimeric IgA, not complexed with the secretory component. They disappeared more rapidly from serum than mIgA antibodies. Anti-V. cholerae IgA responses were parallel in serum and milk samples, whereas anti-S. typhi responses were dissociated. In milk, IgA antibodies were secretory IgA for both antigenic specificities, being probably of local origin in the mammary gland. Our results indicate that both oral and parenteral vaccinations can induce pIgA antibodies in serum and secretions, confirming that the presence of pIgA in serum does not necessarily reflect an immune stimulation only at the mucosal level.

Administration, Oral↗
PubMed2006

A unique set of SH3-SH3 interactions controls IB1 homodimerization.

Islet-brain 1 (IB1 or JIP-1) is a scaffold protein that interacts with components of the c-Jun N-terminal kinase (JNK) signal-transduction pathway. IB1 is expressed at high levels in neurons and in pancreatic beta-cells, where it controls expression of several insulin-secretory components and secretion. IB1 has been shown to homodimerize, but neither the molecular mechanisms nor the function of dimerization have yet been characterized. Here, we show that IB1 homodimerizes through a novel and unique set of Src homology 3 (SH3)-SH3 interactions. X-ray crystallography studies show that the dimer interface covers a region usually engaged in PxxP-mediated ligand recognition, even though the IB1 SH3 domain lacks this motif. The highly stable IB1 homodimer can be significantly destabilized in vitro by three individual point mutations directed against key residues involved in dimerization. Each mutation reduces IB1-dependent basal JNK activity in 293T cells. Impaired dimerization also results in a reduction in glucose transporter type 2 expression and in glucose-dependent insulin secretion in pancreatic beta-cells. Taken together, these results indicate that IB1 homodimerization through its SH3 domain has pleiotropic effects including regulation of the insulin secretion process.

Adaptor Proteins, Signal Transducing↗
PubMed1999

Uterine-associated serine protease inhibitors stimulate deoxyribonucleic acid synthesis in porcine endometrial glandular epithelial cells of pregnancy.

Protease inhibitors are major secretory components of the mammalian uterus that are thought to mediate pregnancy-associated events primarily by regulating the activity of proteolytic enzymes. In the present study, we examined the mitogenic potentials of two serine protease inhibitors, namely secretory leukocyte protease inhibitor (SLPI) and uterine plasmin/trypsin inhibitor (UPTI) in primary cultures of glandular epithelial (GE) cells isolated from early pregnant (Day 12) pig endometrium, using the [(3)H]thymidine incorporation assay. Purified porcine SLPI (pSPLI), porcine UPTI (pUPTI), or recombinant human SLPI (rhSLPI), all of which exhibited anti-trypsin activity, increased (p < 0.05) labeled thymidine incorporation into DNA of serum-deprived GE cells when tested at a range of 10-1000-ng/ml concentrations. Polyclonal antibodies directed against either hSLPI or pSLPI abrogated the effect of SLPI. Co-addition of pSLPI and pUPTI increased DNA synthesis in these cells to a level higher (p < 0.05) than that observed with either protease inhibitor. The glycosaminoglycan heparin, which has been previously shown to increase the anti-protease activity of SLPI, exhibited a tendency (p = 0.08) to enhance SLPI and UPTI induction of cellular DNA synthesis. Reverse transcription-polymerase chain reaction indicated that the messenger RNAs for both protease inhibitors were present in the endometrium throughout pregnancy and, within this tissue, in GE cells to a greater extent (p < 0.05) than in stromal fibroblastic cells. Results demonstrate that, in addition to their well-documented anti-protease activities, SLPI and UPTI may constitute autocrine growth promotants for the uterine epithelium. These data suggest a novel mechanism whereby locally produced protease inhibitors may modulate periimplantation events and embryo-maternal communication.

Animals↗
PubMed2000

Mammary gland immunology and neonate protection in pigs. Homing of lymphocytes into the MG.

Since placenta of pregnant sows are impermeable to immunoglobulin passage, the neonates are born agammaglobulinemic; although immunocompetent, they are unable to develop rapidly an immune response which will protect their systemic and mucosal compartments; thus their survival depend upon the passive acquisition of maternal immunity including at least 3 components: i) a systemic humoral immunity, transmitted through colostrum conveying mainly by IgG; these IgG are transferred from maternal serum via Fc gamma receptors on the epithelial cells of mammary gland (MG). ii) a local humoral immunity, especially secretory IgA (IgAs), transmitted mainly by milk (lactogenic immunity) until weaning. IgAs are secreted by MG recruited plasma cells and are excreted in milk via secretory component of epithelial cells: these IgA exhibit a specificity for the antigens present in the maternal digestive tract, the so-called "entero-mammary link"; this link is due to the migration of lymphocytes from the gut to the mammary gland; they are recruited from the blood via the interaction of their homing receptor (alpha 4 beta 7) with the developmentally regulated mucosal vascular addresin MadCAM-1. In the MG, MadCAM-1 increased in pregnancy (probably under oestrogenic stimulation) but regressed in lactation; its density is closely related to the T cell numbers in MG; in contrast the increase in plasma cell numbers is not related to MadCAM-1 density. Thus IgA precursor cells (alpha 4 beta 7 B cells) seem to be recruited by a milk B cell chemoattractant. On the other hand, presence of T and B lymphocytes in MG (some of them originating from the systemic compartment), sustains the attempts of MG immunization and the results sustain the view of a true local immune response. iii) possibly but not formally proved, a cellular immunity transmitted via maternal immunocompetent cells present in mammary secretions; the exported lymphocytes may represent a selected population of lymphocytes after their passage through the MG epithelium.

Animals↗
PubMed1990

Characterization of ocular mucus extracts by crossed immunoelectrophoretic techniques.

Crossed immunoelectrophoresis (CIE) and crossed immunoelectrofocusing (CIEF) were used to characterize proteins and mucosubstance in the saline extract of human ocular mucus pooled from the normal eyes of donors. CIE resolved more components with greater specificity than previous techniques. Up to 25 components were identified. Lactoferrin, protein G, tear prealbumin, and ocular mucosubstance were found to be ocular-specific. CIE also allowed for the study of protein associations of: 1) albumin-alpha 1-antitrypsin; 2) albumin-tear prealbumin; and 3) IgA-secretory component and lactoferrin-mucosubstance. CIEF revealed that most of the proteins were in the pI range of 4.6-7.4. Up to 19 components were identified. Protein associations revealed by CIE were not evident by CIEF. These results provide a basis for future comparative analyses of tear and mucus from normal and diseased eyes, essential for a better understanding of tear and mucus function.

Adult↗
PubMed2001

Activity of human IgG and IgA subclasses in immune defense against Neisseria meningitidis serogroup B.

Both IgG and IgA Abs have been implicated in host defense against bacterial infections, although their relative contributions remain unclear. We generated a unique panel of human chimeric Abs of all human IgG and IgA subclasses with identical V genes against porin A, a major subcapsular protein Ag of Neisseria meningitidis and a vaccine candidate. Chimeric Abs were produced in baby hamster kidney cells, and IgA-producing clones were cotransfected with human J chain and/or human secretory component. Although IgG (isotypes IgG1-3) mediated efficient complement-dependent lysis, IgA was unable to. However, IgA proved equally active to IgG in stimulating polymorphonuclear leukocyte respiratory burst. Remarkably, although porin-specific monomeric, dimeric, and polymeric IgA triggered efficient phagocytosis, secretory IgA did not. These studies reveal unique and nonoverlapping roles for IgG and IgA Abs in defense against meningococcal infections.

Animals↗
PubMed1975

[Study of human secretory immunoglobulin A. I. Obtaining monospecific antiserum to human secretory immunoglobulin A].

A method of obtaining monospecific antiserum to the human secretory IgA is described. Immunochemically pure secretory IgA (isolated from human colostrum by fractionation with ammonium sulfate and gel-filtration on Sephadex G-200) was used for immunization of rabbits or sheep. Heterologous antibodies were removed by adsorption with commercial gamma globulin, normal serum, the serum of a patient suffering from A-myeloma with the IgA polymere and purified lactoferrin. Monospecific antiserum to the secretory IgA gave a reaction of complete immunological identity with the secretory IgA and a free secretory component.

Adsorption↗
PubMed1988

Complex-type glycoproteins synthesized in the subcommissural organ of mammals. Light- and electron-microscopic investigations by use of lectins.

The secretory activity in the subcommissural organ (SCO) of the sheep and cow was examined by means of lectin histochemistry and cytochemistry. Among the various lectins tested. Concanavalin A (Con A) revealed glycoproteins rich in mannosyl residues in the rough endoplasmic reticulum of ependymal and hypendymal cells. One of these Con A-positive glycoproteins may represent the precursor of the specific secretory component elaborated in the SCO, giving rise to Reissner's fiber. Lens culinaris agglutinin (LCA) and Phaseolus vulgaris hemagglutinins (E-PHA and L-PHA), known to bind to oligosaccharides, as well as wheat-germ agglutinin (WGA) revealing neuraminic acid, labeled secretory granules located in the apical part of ependymal and hypendymal cells of ruminants, and also Reissner's fiber. Electron-microscopic visualization of WGA-positive material in the Golgi complex shows that complex-type glycoproteins are synthesized in the subcommissural organ of mammals. The electron-dense material is mainly secreted into the ventricular cavity and gives rise to Reissner's fiber. On the basis of lectin affinity for oligosaccharides, a structure of the complex-type oligosaccharide is proposed.

Animals↗
PubMed1980

[Amphibian jelly envelopes (proteins secreted by the oviduct) and their role in fertilization].

The jelly envelopes, surrounding amphibian eggs and playing a major part in the initial steps of fertilization, are deposited around the eggs as they traverse the different parts of the oviduct. These envelopes form 5 to 8 morphologically distinct layers according to the species, i.e. in Anura and Caudata they are organized differently, and are composed of about ten different macromolecular components (glycoproteins). The innermost layer is rich in sulfate and poor in fucose, a carbohydrate abundant in the other layers. This is due to a fucosyltransferase deficit in the cranial part of the oviduct secreting this layer. The cranial and the medial oviduct secretory products show a lectin carbohydrate-like reaction which is pH-dependent and can be inhibited by carbohydrates like glucuronolactone. This reaction has made it possible to isolate, by affinity chromatography, the main secretory component of the cranial oviduct (Spop). This paper discusses the main functions of the jelly layers of the egg, with particular reference to fertilization. The role of their macromolecular components in sperm capacitation is clear in Anura; experimental investigation in Urodeles is more difficult. We propose several hypotheses to explain the molecular interactions leading to capacitation. Block to polyspermy in amphibians is discussed.

Amphibians↗
PubMed1977

[Secretory IgA in amniotic fluid].

Immunoglobulin composition was studied in the amniotic fluid (AF) and in the urine of newborn babies. IgA was revealed with the aid of anti-secretory component of the serum in 50 of 57 AF samples; in the urine of babies in the course of the first 24 hours after birth it was revealed in 2 of the 21 cases only. This fact and also the detection of the secretory IgA in the AF at the early periods of pregnancy permit to suppose the synthesis of IgA or of its C-fragment in the amniotic membranes.

Amniotic Fluid↗
PubMed2002

Growth of purified lacrimal acinar cells in Matrigel raft cultures.

The objective of this study was to develop a tissue culture system which closely mimics the in situ lacrimal gland for improved study of lacrimal acinar cell physiology. Highly purified preparations of lacrimal acinar cells from adult female New Zealand White rabbits were isolated and grown in suspension culture in the form of Matrigel 'rafts', i.e., aggregates of acinar cells enclosed within a Matrigel coating. The rafts were seeded onto Matrigel-coated culture plates and their growth was followed for up to 28 days. Immunohistochemistry was used to demonstrate the cellular sites of prolactin (PRL), epidermal growth factor (EGF), basic fibroblast growth factor (FGF-2), secretory component (SC) and major histocompatibility complex class-II molecules (MHC-II) within the acinar cells. By 3 days the cultures contained numerous, well-formed acini enclosed within the Matrigel. The acinar epithelial cells demonstrated histotypic polarity, with large, pale-staining, secretory granules aggregated adjacent to the lumen, and exocytotic release of secretory material into the lumen. From 5-10 days the pale-staining secretory granules decreased in number, while the lumenal contents of the acini increased in staining density. Throughout the culturing period as the pale-staining, secretory granules decreased in number, smaller more densely stained, secretory granules increased in number. The number of cells and size of acinar clusters increased steadily throughout the culturing period, and acini frequently achieved dimensions in excess of 0.5 mm. Increases in the size of acinar clusters were often accompanied by an increase in the size of the lumen. Frequently the lumen and its contents bulged asymmetrically towards one edge of the acinus. Immunhistochemistry demonstrated PRL and EGF within the lumens and within the apical cytoplasm of the acinar cells. Acini were strongly immunopositive for SC throughout the 28 day culture period, whereas immunopositivity for MHC-II molecules was strong initially, but diminished dramatically by 21 days. Immunostaining for FGF-2 was most intense on days 1 and 3, with staining throughout the cytoplasm, but became progressively more localized to the periphery of the acini as the culture period lengthened. In cultures of 1-28 days duration, Western blots of cell lysates demonstrated a major band (approximately 40 kDa) for PRL in 3-28 day preparations; a major band (approximately 80 kDa) for SC in 3 day and 7 day preparations that decreased in intensity in 14-28 day preparations; and a major band (approximately 23 kDa) for MHC-II protein in 1-21 day preparations that decreased in intensity in 28 day preparations. Lysosomes increased in number with time in culture, becoming a dominant cytoplasmic feature in 21 and 28 day cultures. Carbachol stimulation of 4 day rafts resulted in increased release of beta-hexosaminidase and SC from the rafts. The authors conclude that Matrigel rafts containing purified lacrimal gland acinar cells offer a highly advantageous system for study of lacrimal acinar cell function and one that correlates well with the in situ gland.

Animals↗
PubMed1994

99Tcm-cystine, a renal function and imaging agent: a comparative study in dog with 131I-hippurate and 99Tcm-glucoheptonate to evaluate its functional and imaging characteristics.

99Tcm-cystine, which has been proposed as a renal radiopharmaceutical for evaluating renal morphology and function in a single experiment, is compared with 131I-orthoiodohippurate (OIH) with respect to its renal clearance and extraction parameters and with 99Tcm-glucoheptonate (GHA) regarding its imaging characteristics. In spite of its comparable renal accumulation with 131I-OIH, its clearance (10.1 +/- 1.0 ml min-1 kg-1) was lower than that of 131I-OIH (21.5 +/- 0.9 ml min-1 kg-1) but was higher than that of 125I-iothalamate (5.4 +/- 0.6 ml min-1 kg-1). Extraction efficiencies of 99Tcm-cystine, 131I-OIH and 125I-iothalamate were 39 +/- 5, 64 +/- 4 and 27 +/- 3, respectively. The glomerular filtration components of 99Tcm-cystine and 131I-OIH were 26 and 16% of their respective clearances. In probenecid-treated animals the clearance of both agents was affected to a similar extent and fell to half of their respective control values, whereas tubular secretory components were found to be 19 and 31% of the controls. The kidney images obtained with 99Tcm-cystine were superior to those obtained with 99Tcm-GHA at different time points. Therefore, considering both renal function and imaging properties of 99Tcm-cystine it appears that this radiopharmaceutical offers some definite advantages over the currently available renal agents and commands further study.

Animals↗
PubMed1997

Abnormal predominance of IgG in HIV-specific antibodies produced by short-term cultured duodenal biopsy specimens from HIV-infected patients.

The aim of our study was to analyze HIV-specific humoral immunity in the intestinal mucosa at different stages of HIV infection in comparison with serum and saliva. Duodenal biopsy specimens from 30 AIDS patients and 9 HIV-infected patients without AIDS were cultured for 48 hours. Culture supernatants, as well as simultaneously obtained serum and saliva samples, were adjusted to the same immunoglobulin concentrations and tested for HIV-specific IgG and IgA by Western blot. The HIV antigen pattern differed clearly between IgA and IgG but was similar for each isotype independent of its origin (i.e., serum, saliva, or biopsy specimen supernatants). Short-term cultured duodenal biopsy specimens from HIV-infected patients at all stages produced predominantly IgG, which was broadly reactive with HIV antigens. Lower titers of HIV-specific IgA, which recognized few antigens, were found, mostly the glycoprotein gp160. At later stages of the disease compared with earlier stages, the reaction pattern of mucosal IgA from saliva and biopsy supernatants was even more restricted; secretory component was frequently absent. The abnormal predominance of HIV-specific IgG over IgA in mucosal secretions may result from abnormal antibody production in the mucosa rather than from serum leakage. Mucosal inflammation induced by HIV-IgG immune complexes and insufficient immune exclusion by secretory IgA may not only lead to increased mucosal HIV replication but may also contribute to gastrointestinal disease in HIV-infected patients.

Adult↗
PubMed1994

[Relationship between secretory IgA in pancreatic secretions, in pancreatic tissue, and in the serum of patients with chronic pancreatitis].

In five patients with chronic pancreatitis we found secretory IgA (sIgA) in the pancreatic juice. In four control persons this was seen only in one case. In pancreatic tissue of three patients with chronic pancreatitis we detected both, IgA and secretory component by immunohistology. We suggest that sIgA in the pancreatic juice was partly produced in the pancreas.

Chronic Disease↗
PubMed1994

The role of the carbohydrate chains in complement (C3) fixation by solid-phase-bound human IgA.

In contrast to antigen-antibody complexes containing native human IgA, solid-phase-deposited IgA activates the alternative complement pathway and binds C3b. To investigate the role of carbohydrate chains in this, various human IgA preparations were treated with neuraminidase alone or together with N-glycanase or O-glycanase, or with mixed glycosidases from the oral bacterium, Streptococcus mitis. Depletion of oligosaccharides was determined by carbohydrate analysis. Removal of sialic acid and N-linked glycan chains greatly increased the C3b-fixing properties of normal serum IgA1 and IgA2. Myeloma IgA1 and IgA2 proteins and secretory IgA had higher C3b-binding activity than normal serum IgA, and this was further increased by removal of sialic acid and N-linked glycans. Fc alpha and Fc alpha-SC fragments of myeloma and secretory IgA1, respectively, but not Fab alpha fragments, obtained by cleavage with bacterial IgA1 proteases and also free secretory component, fixed C3b by the alternative pathway.

Complement C3b↗
PubMed2006

The B-cell system in inflammatory bowel disease.

Secretory immunity is the best-defined part ot the mucosal immune system. This adaptive humoral defense mechanism depends on a fine-tuned cooperation between secretory epithelia and local plasma cells. Such mucosal immunocytes produce preferentially dimers and larger polymers of immunoglobulin A (collectively called pIgA), which contain J chain and therefore can bind to the epithelial secretory component (SC). This transmembrane glycoprotein functions as pIg receptor (pIgR) that also translocates pentameric IgM to the epithelial surface. B cells with a high level of J-chain expression and pIg-pIgR interactions at mucosal effector sites are thus necessary for the generation of secretory antibodies (SIgA and SIgM). Secretory antibodies perform immune exclusion in a first-line defense, thereby counteracting microbial colonization and mucosal penetration of soluble antigens. However, local production of pIgA is significantly down-regulated in inflammatory bowel disease (IBD), as revealed by strikingly decreased J-chain expression. Although the total increase of the immunocyte population in IBD lesions probably compensates for the relatively reduced pIgA production, decreased pIgR/SC expression in regenerating and dysplastic epithelium signifies that the SIgA system is topically deficient. There is, moreover, a significant shift from IgA2 to IgA1 production, the latter subclass being less resistant to proteolytic degradation. These changes--together with activation of mucosal macrophages and a dramatic increase of IgG-producing cells--may reflect local establishment of a second defense line which, however, is unsuccessful in its attempt to eliminate antigens derived from the indigenous microbial flora. Such a 'frustrated' local humoral immune system results in altered immunological homeostasis and jeopardized mucosal integrity. Complement activation observed in relation to epithelium-bound IgG1 in ulcerative colitis indicates, moreover, that the surface epithelium is subjected to immunological attack by an autoimmune reaction. These luminal deposits regularly contain terminal cytotoxic complement, and often also C3b as a sign of persistent activation. Comparison of identical twins, discordant with regard to ulcerative colitis, suggests that the markedly skewed local IgG1 response seen in this IBD entity may be genetically determined. The initial event(s) eliciting B-cell driven immunopathology in IBD remains unknown. Abrogation of oral tolerance to certain antigens from commensal bacteria has been suggested as a putative early mechanism, and lymphoid neogenesis and hyperplasia in the lesions most likely signify massive microbial overstimulation of the local B-cell system. Such ectopic lymphoid microcompartments may contribute substantially to the proinflammatory systemic-type of B-cell responses occurring in established IBD lesions.

Animals↗