Acetaldehyde induces histamine release from purified rat peritoneal mast cells

Acetaldehyde is a widely distributed compound in the human environment and it is also formed in the human body from various endogenous and exogenous sources, exogenous ethanol being the most important one. Many alcohol-associated hypersensitivity reactions, e.g. Oriental flushing reaction, appear to be attributable to acetaldehyde rather than to ethanol itself. The pathogenetic mechanism behind such hypersensitivity reactions has been suggested to be histamine release from mast cells or blood basophils. However, the direct effects of acetaldehyde on mast cells, the main source of histamine in a mammalian body, have not been studied. The aim of the present study was, thus, to evaluate whether physiological concentrations of acetaldehyde could release histamine from purified rat peritoneal mast cells. The effects of ethanol were studied similarly. The results show that acetaldehyde, already at a concentration of 50 microM, significantly increases the release of histamine from mast cells. Ethanol has a similar effect but only at molar concentrations. These results indicate that acetaldehyde may contribute to the development of various hypersensitivity reactions by directly increasing histamine release from mast cells.     

Identification of a histamine release inhibitory factor (HRIF) and an inhibitor of histamine releasing factor synthesis (IHS) produced by guinea pig lymphoid cells

Guinea pig spleen cells, thymocytes, and blood mononuclear cells have been shown to produce a histamine releasing factor (HRF) spontaneously or after stimulation with PHA or specific antigens. In this study we have shown that antigenic stimulation of spleen cells obtained from animals immunized with high doses of antigen suppresses the spontaneous HRF production. Likewise, stimulation with Con A of spleen cells also inhibits the spontaneous HRF production. When the supernatants from Con A- and antigen-stimulated cultures were added to fresh thymocyte cultures, a significant inhibition of HRF production was observed. These supernatants also inhibited HRF-induced histamine release from mast cells. We have identified an inhibitor of HRF synthesis (IHS) and also a histamine release inhibitory factor (HRIF) by gel chromatography. Virtually all mitogen- and antigen-stimulated culture supernatants elaborated the activities of HRF, IHS, and HRIF in various quantities depending on the dose of antigen and the kind of adjuvant used for immunization. IHS has a MW of 22K-35K and 10K. This cytokine also inhibits DNA synthesis by thymocytes. HRIF has a MW of 15K-20K and 10K. It inhibits both HRF- and antigen-induced histamine release from lung mast cells. These results suggest that lymphocytes produce a variety of factors which function to regulate histamine release from mast cells.     

Cellular origin of histamine-releasing factor produced by peripheral blood mononuclear cells

Histamine releasing factors (HRF) are a group of cytokines that release histamine and other mediators from mast cells and basophils. It has been speculated that HRF might play a major role in the pathogenesis of allergic diseases. Most investigators have studied PBMC as a source of HRF. This study was undertaken to investigate the cellular origin of HRF. Peripheral blood was processed to isolate and purify monocytes, T cells, CD4- T cells, CD8- T cells and B cells by using plastic adherence, 2-aminoethylisothiomonium-treated SRBC rosetting and negative selection with the use of mAb OKM1, OKT11, OKT8, OKT4, and OKB7 plus C. Highly purified subpopulations of PBMC were cultured alone or in the presence of Con A for 24 h. Supernatants were harvested, dialyzed, and assayed for HRF activity in the basophil histamine release test. We found that all subpopulations of PBMC including T cells, CD4- T cells, CD8- T cells, B cells, and monocytes produce variable quantities of HRF. The spontaneous production is very high in B cells but only barely measurable in T cells and monocytes. The synthesis of HRF by B cells was confirmed by abolishing the release of the activity after treatment of B cells with OKB7 mAb and C. Stimulation of cell populations by Con A significantly enhances HRF production by PBMC and T cells but not by B cells and monocytes. In mixing experiments, unstimulated monocytes + B cells showed synergism, but other combinations demonstrated an additive effect. This is the first demonstration of HRF production by human peripheral blood B cells. The results of this study also suggest that histamine releasing cytokines are of multiple cellular origin. This perhaps contributes to their molecular heterogeneity.     

Rapid inhibitory effect of corticosterone on histamine release from rat peritoneal mast cells.

Glucocorticoids are steroids endowed with powerful anti-inflammatory properties, which are routinely believed to require several hours to take effect through modulation of gene expression. Our recent report has shown that glucocorticoids could inhibit allergic reaction within 10 minutes, which the classical genomic mechanism could not explain. Histamine is thought to be one of major mediators in the allergic reaction, and IgE-mediated histamine release from mast cells plays a pivotal role in allergic diseases. Here, we have determined a rapid effect of corticosterone on histamine release from rat peritoneal mast cells, using fluorometric assay. The results showed that corticosterone could inhibit antigen-induced histamine release from rat peritoneal cells within 15 minutes (p<0.05), which could be mimicked by membrane-impermeable BSA conjugated corticosterone (p<0.05). Neither glucocorticoid nuclear receptor antagonist nor protein synthesis inhibitor could block the rapid action (p<0.05). The study provided evidence that nongenomic mechanism might be involved in rapid effect of glucocorticoids on mast cells in allergic disease.     

Effect of highly purified thymus factor on the cellular and humoral indices of immunity in thymectomized mice]

Effect of homogeneous polypeptide thymus factor of mol weight about 5000 (thymarin-III) on cellular and humoral immune responses of thymectomized adult CBA mice was studied. Thymectomy proved to greatly decrease the number of T-cells in the spleen. Accordingly, the capability of these mice to produce both IgM and IgG antibody-forming cells in response to the thymus-dependent antigen (sheep red blood cells) was significantly depressed. Subcutaneous injections of thymarin-III (1 microgram per g of body weight) for 7 days completely restored the T-cell spleen population and normalized the animals' immune response.     

Platelet factor 4 fragment induces histamine release from rat peritoneal mast cells

Platelet factor 4 (PF-4) belongs to a superfamily of low-molecular weight proteins known as chemokines. However, its function has not been fully evaluated. In the present study, we investigated the effect of PF-4 on histamine release from rat peritoneal mast cells by employing its biologically-active carboxyl-terminal fragment, PF-4 (58-70). PF-4 (58-70) stimulated histamine release from mast cells in a dose-dependent manner (10(-8) to 10(-5)M). Histamine release induced by PF-4 (58-70) occurred rapidly (<30s) and was inhibited by extracellular Ca(2+). These results suggest that PF-4 might play a crucial role at the site of inflammation and/or immune response.     

Preparation of highly purified F-actin-depolymerizing factor of human serum

F-actin depolymerizing factor (ADF) of human serum was purified 250-400-fold to more than 98% purity with high reproducibility. The purification included (1) 30-50% (NH4)2SO4 precipitation, (2) ion-exchange chromatography on DEAE-Sepharose, (3) chromatofocusing on Polybuffer exchanger 94 and (4) affinity chromatography on ConA-Sepharose. The recovery of ADF was estimated to be 20-30% whereas the ADF activity yield was 5-17%. The lower activity yield was thought to be due-partly to proteolysis and partly to destabilization of highly purified ADF.     

Induced release and metabolism of arachidonic acid from myeloid cells by purified colony-stimulating factor

The in vitro incubation of cells from turpentine-induced rat myeloid hyperplastic marrow and peritoneal monocyte/macrophage with 14C-arachidonic acid resulted in the incorporation of the radiolabel into the particulate phospholipids. Challenge of the radiolabeled cells with a highly purified type I CSF (CSF I) from human pancreatic carcinoma cells in continuous culture resulted in the hydrolysis and release of the 14C-arachidonic acid from the cellular phospholipids. The simultaneous challenge of the prelabeled cells with CSF-I and its specific antibody (anti-CSF-I antibody) inhibited the CSF-I induced hydrolysis of 14C-arachidonic acid from the cells. These results confer a specificity on the CSF-I induced release of arachidonic acid from the cellular phospholipids. Our data also demonstrated that the 14C-arachidonic acid released from the cellular phospholipids was further transformed into products of the cyclooxygenation and lipoxygenation pathways by cellular enzyme systems in both populations of cells. Interestingly, our data also indicate that the challenge of the granulocytic hyperplastic marrow cells and the monocyte/macrophage cells with purified CSF-I resulted in a higher generation of lipoxygenase products in the predominantly granulocytic cell population than in the population rich in monocyte/macrophage cells. The biological significance of this observation remains to be further explored. Thus, the CSF-I induced release of cellular arachidonic acid explains, at least in part, the presence of prostaglandins and other metabolites of arachidonic acid that are found in the media of hemopoietic cells incubated with a variety of CSF preparations.     

Cellular origin and release of intrinsic factor from isolated rat gastric mucosal cells

The cellular content and secretion of intrinsic factor was measured by [57Co]cyanocobalamin binding using isolated rat gastric mucosal cells. The intrinsic factor/R-protein ratio was above 9:1 as evaluated by specific anti-intrinsic factor antibodies. In unfractionized cells with 23 +/- 1.3% parietal cells the intrinsic factor content of 148 +/- 47 fmol/10(6) cells remained almost unchanged over 3 h, whereas basal secretion rose up to 57 +/- 10. In fractionized cells (Percoll) with 3-85% parietal cells most intrinsic factor was found in the parietal cell-depleted fraction (content: 441 +/- 30, secretion/3 h: 139 +/- 16, mean formation/h: 50 +/- 12 fmol/10(6) cells). The intrinsic factor content of the different cell fractions correlated with that of pepsin. [14C]Aminopyrine uptake, an indirect measure of parietal cell H+ production, was inversely related. Carbachol (1 X 10(-6)-10(-3) mol/l) stimulated intrinsic factor secretion, 1 X 10(-3) mol/l being maximally effective (90 +/- 8% above basal). This response was inhibited by atropine and pirenzepine, but not by prostaglandin E2 (PGE2) and somatostatin. Dibutyryl cyclic adenosine monophosphate (dibutyryl cAMP, 43 +/- 7%) and hexoprenaline (24 +/- 5%) enhanced intrinsic factor secretion less effectively and pentagastrin like histamine lacked any stimulatory effect. We conclude that in the rat intrinsic factor is produced and released from chief cells mainly under cholinergic control.     

Cellular origin and release of intrinsic factor from isolated rat gastric mucosal cells

The cellular content and secretion of intrinsic factor was measured by [⁵⁷Co]cyanocobalamin binding using isolated rat gastric mucosal cells. The intrinsic factor/R-protein ratio was above 9:1 as evaluated by specific anti-intrinsic factor antibodies. In unfractionized cells with 23 ± 1.3% parietal cells the intrinsic factor content of 148 ± 47 fmol/10⁶ cells remained almost unchanged over 3 h, whereas basal secretion rose up to 57 ± 10. In fractionized cells (Percoll^®) with 3–85% parietal cells most intrinsic factor was found in the parietal cell-depleted fraction (content: 441 ± 30, secretion/3 h: 139 ± 16, mean formation/h: 50 ± 12 fmol/10⁶ cells). The intrinsic factor content of the different cell fractions correlated with that of pepsin. [¹⁴C]Aminopyrine uptake, an indirect measure of parietal cell H⁺ production, was inversely related. Carbachol (1·10⁻⁶−10⁻³ mol/l) stimulated intrinsic factor secretion, 1·10⁻³ mol/l being maximally effective (90 ± 8% above basal). This response was inhibited by atropine and pirenzepine, but not by prostaglandin E₂ (PGE₂) and somatostatin. Dibutyryl cyclic adenosine monophosphate (dibutyryl cAMP, 43 ± 7%) and hexoprenaline (24 ± 5%) enhanced intrinsic factor secretion less effectively and pentagastrin like histamine lacked any stimulatory effect. We conclude that in the rat intrinsic factor is produced and released from chief cells mainly under cholinergic control.     

Nipah virus infects specific subsets of porcine peripheral blood mononuclear cells

Nipah virus (NiV), a zoonotic paramyxovirus, is highly contagious in swine, and can cause fatal infections in humans following transmission from the swine host. The main viral targets in both species are the respiratory and central nervous systems, with viremia implicated as a mode of dissemination of NiV throughout the host. The presented work focused on the role of peripheral blood mononuclear cells (PBMC) in the viremic spread of the virus in the swine host. B lymphocytes, CD4-CD8-, as well as CD4+CD8- T lymphocytes were not permissive to NiV, and expansion of the CD4+CD8- cells early post infection was consistent with functional humoral response to NiV infection observed in swine. In contrast, significant drop in the CD4+CD8- T cell frequency was observed in piglets which succumbed to the experimental infection, supporting the hypothesis that antibody development is the critical component of the protective immune response. Productive viral replication was detected in monocytes, CD6+CD8+ T lymphocytes and NK cells by recovery of infectious virus in the cell supernatants. Virus replication was supported by detection of the structural N and the non-structural C proteins or by detection of genomic RNA increase in the infected cells. Infection of T cells carrying CD6 marker, a strong ligand for the activated leukocyte cell adhesion molecule ALCAM (CD166) highly expressed on the microvascular endothelial cell of the blood-air and the blood-brain barrier may explain NiV preferential tropism for small blood vessels of the lung and brain.     

Inhibitory effect of diethylstilbestrol on histamine release by rat mast cells and its relation to the cellular ATP content

The effect of diethylstilbestrol, a synthetic estrogen, on mast cell secretion was investigated. The results showed that 50 microM diethylstilbestrol inhibited histamine release from rat peritoneal mast cells in the presence and absence of glucose, but did not affect 45Ca uptake stimulated by concanavalin A. Diethylstilbestrol also inhibited histamine release induced by compound 48/80, exogenous ATP, or ionophore A23187. Since estradiol benzoate, hexestrol and daidzein were not inhibitory, the inhibitory action of diethylstilbestrol must be independent of its estrogenic activity. The ATP content of mast cells decreased to less than 0.1 nmol/10(6) cells on treatment with 50 microM diethylstilbestrol at 37 degrees C for 15 min. This effect of diethylstilbestrol in decreasing the ATP content of mast cells correlated well with its inhibitory effect on histamine release. Diethylstilbestrol at 50 microM depleted the cells of ATP at 37 degrees C, but not at 0 degrees C, whereas [3H]diethylstilbestrol ( [monoethyl-3H]diethylstilbestrol) binding to rat mast cells was the same at 0 and 37 degrees C. It is concluded that diethylstilbestrol reduced the ATP content of rat mast cells by inhibiting metabolism of the cells, and consequently inhibited degranulation.     

IgE-mediated release of histamine from human cutaneous mast cells

We investigated the ability of antigen-IgE interactions to stimulate histamine release from human infant cutaneous mast cells. Skin obtained at circumcision contained numerous perivascular mast cells, as assessed by light and electron microscopy. The histamine content of this tissue averaged 17.7 ng (+/- 1.5 SEM)/mg wet weight. Challenge of 200-microns thick sections of unsensitized skin with varying concentrations of monoclonal murine antibodies to human IgE caused no net release of histamine. After skin sections were incubated in the presence of 5 micrograms/ml of human myeloma IgE (S) for 120 min at 37 degrees C, monoclonal anti-IgE challenge resulted in 40.1% (+/- 6.0 SEM) histamine release. Similar passive sensitization with 1/20 dilutions of serum from humans expressing IgE to purified Juniperus sabinoides (JS) antigen rendered the tissue responsive to specific antigen challenge. Dose-related histamine release occurred over 30 min with optimal release of 12.6% (+/- 2.4 SEM) after stimulation with 100 ng/ml of JS antigen. This reaction required sensitization with serum containing IgE to JS and was antigen-specific. Optimal reactions to antigen occurred at 3 mM added Ca++, 34 degrees C to 37 degrees C, pH 7.2. Antigen-induced release was markedly influenced by the added Ca++ concentration; no release occurred in the absence of Ca++, 54% of the optimal response was observed at 2 mM Ca++, and 28% of the optimal response occurred at 4 mM Ca++. The addition of Mg++ did not influence antigen-induced release. The results of this study provide functional evidence that 1) human infant cutaneous mast cells express Fc-epsilon receptors; 2) these receptors are largely unoccupied in vivo; and 3) stimulation of passively sensitized infant mast cells with anti-IgE or specific antigen leads to immediate histamine release. This new system should permit detailed in vitro studies of immediate hypersensitivity reactions in human skin.     

Induction of release of tumor necrosis factor and IL-6 from human mononuclear cells by Bacteroides strains

The role of anaerobic Gram-negative bacteria in inducing cytokines during mixed infections involving aerobic and anaerobic bacteria is relatively poorly defined. The purpose of this study was to establish whether or not intact Bacteroides fragilis and related species, isolated from severe infections and from the faeces of healthy persons are capable of releasing tumor necrosis factor (TNF) and IL-6 from human mononuclear cells and whole blood. The purified lipopolysaccharides of Bacteroides fragilis strain (No. 7), extracted by the aqueous phenol method from BHI cultures and from BHI culture supplemented with 5% horse serum, were also tested. TNF release was detected by the WEHI 164-dependent bioassay and IL-6 production by the B-9 cell-dependent bioassay. Heat-inactivated Bacteroides strains belonging to different species were able to induce TNF (1x10(1)-5x10(2) U/mL) and IL-6 (1x10(1)-5x10(5) pg/mL) release from human mononuclear cells. When whole blood was used, the production of TNF and IL-6 was more pronounced (very probably because of the presence of certain serum factors). The culturing conditions (the presence of 5% horse serum in the BHI broth) influenced the inducing activity of almost all strains tested. The isolated lipopolysaccharide of Bacteroides fragilis strain No. 7 proved to have a rough profile on PAGE. There were no differences in TNF and IL-6 induction when the lipopolysaccharides of the strain was cultured in BHI or in BHI supplemented with 5% horse serum. Bacteroides strains often outnumber Enterobacteriaceae in the faeces and in mixed infections, and their role in inducing and/or modulating the host response in septic shock should not be overlooked.     

Glycosylation of interleukin-6 purified from normal human blood mononuclear cells.

Interleukin 6 (IL-6) is a glycosylated cytokine which is important in exerting cell-specific growth-inducing, growth-inhibiting and differentiation-inducing effects. IL-6 produced in mammalian cell lines is heterogeneous, reflecting specific cell-type-dependent post-translational modifications. Native IL-6 was purified from human blood mononuclear cells and the oligosaccharides released, radiolabelled and sequenced by a combination of sequential exoglycosidase digestion using Bio-Gel P-4 high-resolution gel chromatography and acetolysis. N- and O-linked glycans were found. The N-linked glycans were sialylated di- and tri-antennary complex-type and oligomannose-type structures. However, the most predominant N-linked oligosaccharide was a small tetrasaccharide with the sequence Man alpha 6Man beta 4GlcNAc beta 4GlcNAc. This is the first report of this structure on a circulating glycoprotein. This structure has only previously been reported to be present on the syncytiotrophoblast of human placenta. The presence of the oligomannose structures and the mannose-terminating tetrasaccharide on IL-6 may be important in maintaining a high local concentration of the cytokine while limiting its systemic serum level via interaction with soluble mannose-binding serum lectins.     

Lack of cellular cytotoxicity by human mononuclear cells to Giardia

Cytotoxicity of mononuclear leukocytes (MNL) to Giardia lamblia was compared by using two techniques. The first method assessed the viability of surviving Giardia directly by culturing and the second method measured release of incorporated [3H] thymidine. Cytotoxicity, as measured directly by culturing and visual assessment, showed that the numbers of surviving Giardia decreased over time whether cultured with or without MNL but that Giardia survived significantly better in the presence of MNL at 18 hr (21.8 +/- 8.3% with MNL compared with 3.4 +/- 1.5% without MNL). Release of [3H]thymidine increased whether Giardia were cultured with or without MNL and although there was a tendency for increased release with MNL, there was no significant difference. Dead labeled Giardia released significantly more label in the presence of MNL than without MNL, suggesting that MNL cause release of [3H]thymidine after phagocytosis. The thymidine release assay therefore does not measure spontaneous cytotoxicity of MNL to Giardia.