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Biomedical subjects

A Hein

Publications and source records attributed to A Hein.

At least 55 records · Page 3Linked to original sources
PubMed1987

Biochemical and morphological characterization of basophilic leukocytes from two patients with myelogenous leukemia.

Basophilic leukocytes from two patients with myelogenous leukemia were enriched to a purity of 10 to 45% by density gradient centrifugation. Ultrastructurally, these basophilic leukocytes contained segmented nuclei and granules with reticular patterns resembling those of normal basophils, and other granules with scroll and grating patterns resembling those of normal connective tissue mast cells. The 35S-labeled macromolecules isolated from these cells were approximately 140,000 m.w. Pronase-resistant proteoglycans bearing approximately 15,000 m.w. glycosaminoglycans. On incubation with chondroitinase ABC, nitrous acid, and heparinase, the 35S-labeled proteoglycans were degraded 50 to 84%, 16 to 43%, and 8 to 37%, respectively, indicating the presence of both chondroitin sulfate and heparin. As assessed by high performance liquid chromatography, the 35S-labeled chondroitin sulfate disaccharides liberated by chondroitinase ABC treatment were approximately 95% monosulfated chondroitin sulfate A and approximately 5% disulfated chondroitin sulfate E. The presence of heparin was confirmed by two-dimensional cellulose acetate electrophoresis of the 35S-labeled glycosaminoglycans. Cell preparations, enriched to 75% basophilic leukocytes by sorting for IgE+ cells, also synthesized 35S-labeled proteoglycans containing chondroitin sulfate and heparin. In one experiment, treatment of the cells with 1 microM calcium ionophore A23187 resulted in a 12% net release of both chondroitin sulfate and heparin containing 35S-labeled proteoglycans, a 57% net release of histamine, and the de novo generation of 8, 8, and 0.16 ng of immunoreactive equivalents of prostaglandin D2, leukotriene C4, and leukotriene B4, respectively, per 10(6) cells. Because only mast cells have been found to contain Pronase-resistant heparin proteoglycans, to generate PGD2 on cell activation, and to contain granules with scroll and grating patterns, these findings indicate that in some patients with myelogenous leukemia there are basophilic cells that possess properties of tissue mast cells.

Arachidonic Acid↗
PubMed1987

Effects of iron-, manganese-, or magnesium-deficiency on the growth and morphology of Euglena gracilis.

Iron-, manganese-, or magnesium-deficiency has been induced in Euglena gracilis. Each arrests cell proliferation, decreases the intracellular content of the deficient metal, and increases that of several other metals. Light and electron microscopy of stationary phase cells reveal that Fe-deficient (-Fe) cells are similar in size and shape to control organisms. Magnesium-deficient (-Mg) cells, however, are larger, and approximately 14% are multilobed, containing 2 to 12 lobes of equal size emanating from a central region. Individual (-Mg) cells and each lobe of multilobed cells contain a single nucleus. Manganese-deficient (-Mn) organisms are morphologically more heterogeneous than (-Fe) or (-Mg) cells. Most are spherical and larger than controls. Approximately 15% are multilobed but, unlike (-Mg) cells, contain lobes of unequal size with either zero, one, or several nuclei present in each. Nuclei of (-Mn) cells differ in size and shape from those of control, (-Fe), or (-Mg) cells. All three deficient cell types accumulate large quantities of paramylon. Other cytoplasmic structures, however, appear normal. Addition of Fe, Mn, or Mg to the respective deficient stationary phase cultures reverses growth arrest and restores normal morphology. The results suggest that Fe-, Mn-, and Mg-deficiencies affect different stages of the E. gracilis cell cycle.

Cell Nucleus↗
PubMed1987

Ultrastructural evidence that the granules of human natural killer cell clones store membrane in a nonbilayer phase.

Electron-microscopic examination of five LGL clones, JT3, JTB18, CNK6, CNK7, and CNK10, expressing natural killer activity and T11 and NKH1 phenotype, showed that three of the clones, JT3, CNK6, and CNK7, had crystalline structures in their densest granules. These structures generally consisted of hexagonally packed lattices with a 6.9-nm point-to-point spacing. JTB18 and CNK10 had no structures in their granules. The attack of one clone, JT3, on two resistant target tumor cell lines, KG1 and Laz509, was also examined under three conditions. First, JT3 cells and targets were incubated together. There was little adherence, degranulation, or killing. Second, cells were incubated with anti-T11(2) and T11(3), antibodies against the E-rosette receptor/antigen complex, which activate resting T cells and enhance cytolytic activity of NK clones and CTL. JT3 cells adhered to the targets, formed zones of apposition between NK and target cell membranes, polarized, and degranulated into the space between the two cells, killing the targets. Third, cells were incubated with both anti-T11(2/3) and anti-LFA-1, an antibody that inhibits adherence. The JT3 cells did not form zones of apposition with the targets, but degranulated in discrete areas on their own surface. In all cases, discharged crystalline granules transformed to sheets of membrane and vesicles. These studies suggest that phospholipids are packed in hexagonal lattices in the granules of the resting cells and transform to bilayer structures during exocytosis. The crystalline nature of the granule may immobilize lytic molecules and protect the resting cell from lysis. Further, the vesicles may serve to transport the lytic molecules from the effector to the target cell. Anti-LFA-1 does not inhibit target recognition or exocytosis, but instead blocks membrane interactions of the effector cell with its target.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1986

Inhibition of post-translational modification and surface expression of a melanoma-associated chondroitin sulfate proteoglycan by diethylcarbamazine or ammonium chloride.

Cultured human melanoma M21 cells were treated with diethylcarbamazine (DEC), an inhibitor of proteoglycan biosynthesis in rat chondrosarcoma cells, to examine the assembly and transport of a chondroitin sulfate proteoglycan to the plasma membrane. Pretreatment of melanoma cells at 37 degrees C for 15 min with increasing doses of DEC followed by a 60-min pulse with [35S]sulfate in the presence of DEC resulted in a dose-related inhibition of incorporation of [35S]sulfate into macromolecules. In cells incubated for 75 min with both 1 mM beta-D-xyloside and 15 mM DEC, synthesis and secretion of beta-D-xyloside-bound 35S-glycosaminoglycans were inhibited by more than 80% as compared to cells treated with beta-D-xyloside alone; this inhibition was reversible. As assessed by [3H]serine incorporation into protein, overall protein synthesis was not substantially inhibited by DEC treatment. Detergent lysates from [35S]methionine-labeled melanoma cells were incubated with a monoclonal antibody (9.2.27) that specifically recognizes the peptide core of the melanoma proteoglycan. As assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the immunoprecipitate, a 240,000 Mr endoglycosidase H (Endo-H)-sensitive intermediate was the only form of the proteoglycan present inside the cells when the cultures were treated for 60-120 min with 10-15 mM DEC. When the melanoma cells were incubated for 10 min with 15 mM DEC and 100 mu Ci/ml of [35S]methionine, washed, and then chased for 15 min to 4 h in radioactive-free medium, the 240,000 Mr Endo-H-sensitive intermediate was slowly converted to a 250,000 Endo-H-resistant intermediate but not to a mature proteoglycan molecule that possessed chondroitin sulfate glycosaminoglycans. SDS-PAGE analysis of cell surface immunoprecipitates revealed that only a small amount of the 250,000 Mr intermediate was transported to the plasma membrane within 5 h of incubation in the presence of DEC. Proteoglycan synthesis was also inhibited when the melanoma cells were incubated for 60-120 min with ammonium chloride, but unlike DEC-treated cells the majority of the synthesized peptide core was converted to a 245,000 Mr Endo-H-resistant intermediate that was detected on the cell surface. Light and electron microscopic analysis of DEC-treated melanoma cells revealed large vacuoles and a distended Golgi and endoplasmic reticulum. Ammonium chloride-treated cells contained fewer vacuoles than DEC-treated cells but more vacuoles than normal cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Ammonium Chloride↗
PubMed1986

Biochemical and phenotypic characterization of human basophilic cells derived from dispersed fetal liver with murine T cell factors.

Metachromatically granulated cells were generated from human fetal liver stem cells cultured in heterologous mouse conditioned medium rich in interleukin 3. After 2 to 3 wk of culture with biweekly changes of medium and selection of nonadherent cells, all cells present in five cultures had cytoplasmic granules, and 60 to 95% of the cells stained metachromatically with toluidine blue or with alcian blue but not with the safranin counterstain. Ultrastructurally, many granules contained fibrillar material or electron-dense cores with fibrils and vesicular fragments. In addition, the granules of many cells were filled with electron-dense material, which in some cases had a fine structure consisting of concentric whorls or a reticular pattern. Analysis of high-affinity IgE receptors on the cultured cells by flow cytometry demonstrated a unimodal fluorescence pattern, suggesting that most cells were in the basophil or mast cell lineage. The cultured cells lacked the lymphoid cell surface determinants B1, B4, T3, and T11, the myeloid determinants Mo2 and MY9, the natural killer cell determinant 901, and Ia histocompatibility antigens, but expressed the myeloid determinant MY7. The cells contained 52 ng/10(6) cells of histamine and incorporated [35S]sulfate at an average rate of 31,300 cpm/10(6) cells/4 hr into 175,000 m.w. chondroitin sulfate A proteoglycans. Upon activation with 1 microM calcium ionophore A23187, the cultured cells released 53% of their cell-associated histamine and metabolized arachidonic acid to 15.0 ng/10(6) cells of immunoreactive leukotriene C4 equivalents, 0.5 ng/10(6) cells of leukotriene B4, and 3.1 ng/10(6) cells of prostaglandin D2 (means, n = 3). Thus, stem cells present in human fetal liver give rise, as do stem cells in mouse fetal liver, to metachromatically granulated cells when cultured in the presence of mouse interleukin 3. In both species, the cultured cells bear IgE receptors, lack characteristic lymphoid and most myeloid cell surface determinants, and contain histamine and chondroitin sulfate proteoglycans. The human fetal liver-derived cells are similar in morphology and T cell factor dependence to basophil-like cells derived from umbilical cord blood, but are novel in their capacity to generate leukotrienes and prostaglandin D2.

Animals↗
PubMed1986

[Equivalence of oral and intramuscular premedication. III. Effect of premedication on anesthesia and postoperative pain].

600 patients were given 6 different premedications in randomised design to study their effect on the course of anaesthesia and on postoperative pain. Premedication acts indirectly on anaesthesia, depending on the influence of the drug on anxiety and on the somatic correlates of anxiety. The greater the sedative-anxiolytic effect of the premedication, the easier it is to induce anaesthesia, and the more superficial the anaesthesia, resulting in earlier and stronger onset of postoperative pain. On the other hand, the more anxious the patient is, the more he consumes anaesthetic drugs, whereas anaesthesia remains superficial with the same consequences in respect of postoperative pain. In view of postoperative pain, fast and early awakening from anaesthesia must not be aimed at, particularly after operations which definitely result in postoperative pain (long-term operations in those regions of the body that cannot be immobilised).

Administration, Oral↗
PubMed1986

[Vibration of neck muscles changes the apparent position of a visual target].

The discharge rate of muscle spindle afferents normally provides a precise signal of muscle length. Vibration of a muscle or its tendon induces an increase in afferent discharge which then no longer represents true muscle length; however, this increased proprioceptive input is interpreted in the central nervous system as a lengthening of the muscle. The incremented signal gives rise to illusions of displacement, or movement, of a fixed, vibrated limb. A visual target attached to such a vibrated limb also appears to move. We now report that vibration of the neck muscles influences visual localisation by inducing illusory movement of targets in visual space. Subjects were seated in a totally dark room and viewed a light-emitting diode (LED). The LED was placed at eye level approximately in the body midline at a distance of 70 cm. They held a physiotherapy vibrator in the left hand with its tip against the left side of the neck. When vibration was initiated the LED appeared to move rightward. The position of the tip of the vibrator was adjusted to produce the maximum apparent displacement to the right. In some subjects the illusion had a vertical component. Subjects maintained the vibrator in position and described the illusion when vibration began, during vibration and at its end. They reported that, initially, the target moved to the right but this displacement ceased after a second or two. The target then appeared to continue in motion without changing its position. When vibration ended the target returned to its initial position.(ABSTRACT TRUNCATED AT 250 WORDS)

Central Nervous System↗
PubMed1986

[Equivalence of oral and intramuscular premedication. I. Oral versus intramuscular premedication].

In a randomised study, 6 different premedications (oral flunitrazepam, oral pentobarbital, oral NaCl solution; intramuscular pethidine, intramuscular droperidol/fentanyl, and intramuscular NaCl solution) were administered to 600 patients, to determine the equal efficacy of oral and intramuscular application. The results show a significantly better mode of action of oral premedication.

Administration, Oral↗
PubMed1986

[Equivalence of oral and intramuscular premedication. II. Effect of various conventional premedication drugs].

In a randomised study, 6 different premedications (oral flunitrazepam, oral pentobarbital, oral physiological NaCl solution, intramuscular pethidine, intramuscular droperidol/fentanyl, intramuscular physiological NaCl solution) were administered to 600 patients to determine the equal efficacy of the premedicative drugs in oral or parenteral administration. Patients gave the best rating to the oral application of flunitrazepam. The differences in the assessment of application and effect of the drugs are contrasted by the statement that most patients do not feel any direct influence on their preoperative anxiety by premedication. However, it is easier for them to cope with anxiety because premedication pacifies the patients, whereas each of the dependent variables, such as apprehension, is influenced differently. Oral premedication again proves superior to parenteral premedication.

Administration, Oral↗
PubMed1985

Proteoglycans in cell-mediated cytotoxicity. Identification, localization, and exocytosis of a chondroitin sulfate proteoglycan from human cloned natural killer cells during target cell lysis.

A clone of natural killer (NK) cells (JTB18) was found to be ultrastructurally similar to peripheral blood large granular lymphocytes (LGL). These cells incorporated [35S]sulfate into cell-associated proteoglycan molecules, which were then isolated by CsCl density gradient centrifugation. As assessed by gel filtration chromatography, the native 35S-labeled proteoglycan and its beta-eliminated 35S-labeled glycosaminoglycans were of Mr approximately 200,000 and 50,000, respectively. The 35S-labeled proteoglycans were resistant to proteolysis, since their Mr were apparently not altered by incubation with either pronase or S. aureus V8 protease. The purified NK cell 35S-labeled proteoglycans were degraded by approximately 90% to 35S-labeled disaccharides with either chondroitinase ABC or AC. High performance liquid chromatographic analysis of the digests revealed these disaccharides to be composed entirely of chondroitin sulfate A (glucuronic acid----N-acetylgalactosamine-4SO4). Whole 35S-labeled cells incubated with chondroitinase ABC failed to release 35S-labeled disaccharides into the supernatant, and x-ray energy-dispersive analysis revealed that sulfur-containing molecules were present in the intracellular granules, thereby localizing the NK cell-associated proteoglycan primarily in the granules of the cell, rather than on the plasma membrane. The 35S-labeled cloned NK cells incubated for 30 min to 4 h with K562 tumor cell targets at a 0.5:1 ratio exocytosed a mean of 49% of the granular 35S-labeled proteoglycans during the first 60 min of the culture. Proteoglycan release was maximal with an effector/target cell ratio of 0.5:1 for JTB18:K562. Significant proteoglycan release from JTB18 NK cells was also obtained with other sensitive target cells such as REX, Molt4, and CEM, but not with cells such as KG1 and Laz156, which have been shown previously to be resistant to killing by this NK cell. Thus, protease-resistant intracellular proteoglycans with chondroitin sulfate A side chains are specifically exocytosed from the granules of human NK effector cells upon contact with sensitive targets, suggesting that these proteoglycans may be involved in the mechanism of cytotoxicity.

Cell Line↗
PubMed1985

Novel inhibition of proteoglycan synthesis and exocytosis by diethylcarbamazine in the Swarm rat chondrocyte.

Pretreatment of cultured chondrosarcoma chondrocytes at 37 degrees C for 15 min with 15 mM diethylcarbamazine (DEC) followed by a 60-min pulse with [35S] sulfate in the presence of DEC resulted in an approximate 40% inhibition of synthesis and a 75% inhibition of secretion of 35S-proteoglycan. The inhibition was dose-related and was not due to a decrease in protein synthesis. Chondrocytes exposed for 75 min to 15 mM DEC, washed, incubated for 17 h in DEC-free medium, and then pulsed with [35S]sulfate showed no inhibition in the rate of synthesis of proteoglycan or in the per cent of radiolabeled proteoglycans exocytosed into the culture medium, indicating full reversibility of the inhibitory effect. When chondrocytes were incubated for 75 min with both 1 mM beta-D-xyloside and 15 mM DEC, secretion of beta-D-xyloside-bound 35S-glycosaminoglycan was inhibited by more than 70% despite an approximate 3-fold increase in intracellular 35S-macromolecules, as compared to cells exposed to beta-D-xyloside alone. Upon removal of DEC, the block in the secretion of beta-D-xyloside-bound 35S-glycosaminoglycans was reversed, although there was a 15-30-min lag in the initiation of exocytosis. Light and electron microscopic examination of chondrocytes after 75 min of incubation with 15 mM DEC revealed large vacuoles, a distended Golgi apparatus, and a distended endoplasmic reticulum which contained electron dense material. Upon removal of DEC, the vacuoles disappeared and distended organelles returned to their normal appearance between 15 and 30 min, coincident with the start of exocytosis of 35S-proteoglycan and beta-D-xyloside-bound 35S-glycosaminoglycan. These biochemical and morphological studies indicate that DEC treatment of chondrosarcoma chondrocytes alters the transport of molecules from the endoplasmic reticulum to the Golgi and the transport of molecules from the Golgi to the cell surface.

Animals↗
PubMed1985

Homology of the rat basophilic leukemia cell and the rat mucosal mast cell.

Secretory granules of the rat basophilic leukemia (RBL-1) cell, a chemically generated tumor cell line maintained in tissue culture, were shown to stain with alcian blue but not with safranin counterstain and to have sparse, small, electron-dense granules. A Mr 25,000 protein was the major [3H]diisopropyl fluorophosphate-binding protein in extracts of RBL-1 cells. Double-immunodiffusion analysis of extracts revealed immunoreactivity for rat mast cell protease (RMCP)-II, a Mr 25,000 neutral protease present in the secretory granules of rat mucosal mast cells and cultured rat bone marrow-derived mast cells, but no immunoreactivity for RMCP-I, the predominant neutral protease of rat connective tissue mast cells. By radial immunodiffusion, there was 66.8 ng of RMCP-II per 10(6) cells. Whereas rat connective tissue mast cells stain with alcian blue and safranin and contain heparin proteoglycan, rat mucosal and rat bone marrow-derived mast cells stain with alcian blue only and contain a non-heparin proteoglycan and lesser amounts of histamine. Proliferation of rat mucosal mast cells in vivo and rat bone marrow-derived mast cells in vitro requires T-cell factors, whereas no comparable requirement has been observed for connective tissue mast cells. The transformed RBL-1 tumor cells, whose growth is independent of factors other than those present in standard tissue culture medium, has previously been shown to contain predominantly chondroitin sulfate di-B proteoglycans and low amounts of histamine. The similar histology and secretory granule biochemistry of the rat mucosal mast cells, rat culture-derived mast cell, and RBL-1 cell suggest that they comprise a single mast cell subclass distinct from the rat connective tissue mast cell.

Animals↗
PubMed1985

Fibroblasts maintain the phenotype and viability of the rat heparin-containing mast cell in vitro.

Rat serosal heparin-containing mast cells (HP-MC) were maintained in vitro for as long as 30 days when co-cultured with mouse skin-derived 3T3 fibroblasts. In contrast, when the mast cells were cultured alone, on fibronectin-, gelatin-, or dermal-collagen-coated dishes, on acid and heat-killed fibroblasts in the presence or absence of 24 hr fibroblast-conditioned medium, or on a monolayer of mouse serosal macrophages, they failed to adhere to the dishes, released significant amounts of their histamine and lactate dehydrogenase, and stained with trypan blue, indicating a loss of viability. The rat serosal HP-MC cultured with the 3T3 fibroblasts became so adherent to the fibroblasts that the two cell types could be separated from one another only by trypsinization. The cultured HP-MC stained with both alcian blue and safranin and continued to synthesize proteoglycan at a rate comparable to that of freshly isolated cells. The 35S-labeled proteoglycan synthesized by these cultured cells, like that produced by freshly isolated rat serosal HP-MC, was a 750,000 to 1,000,000 m.w. proteoglycan containing only heparin glycosaminoglycans of 50,000 to 100,000 m.w. When HP-MC were cultured for 1 wk with the fibroblasts and were then incubated for 5 min with a 1/20 dilution of rabbit anti-rat IgE, they generated and released an average of 22 +/- 10 ng (mean +/- SD, n = 5) of prostaglandin D2 per 10(6) cells and exocytosed a higher net percentage of their total histamine content (44 +/- 11% [mean +/- SD, n = 8]) than did cells just isolated from the animal (6 +/- 4% [mean +/- SD, n = 4]). As assessed by electron microscopy, many of the cultured HP-MC resembled freshly isolated cells except that some secretory granules had fused with one another in some cells. Morphologically, after activation the cultured HP-MC underwent compound exocytosis like freshly isolated cells. These results demonstrate that the in vivo differentiated rat HP-MC maintain their histology, morphology, immunologic responsiveness, histamine content, and ability to synthesize heparin proteoglycan when co-cultured with living fibroblasts.

Animals↗
PubMed1984

Anoxic cell swelling dissociated from excess permeability by poorly penetrating solutes.

Increased plasma membrane permeability, cell swelling, and cell death are consequences of metabolic energy deprivation. Rabbit retinas were incubated in an oxygenated artificial cerebrospinal fluid containing glucose. The incubating medium was modified by causing anoxia and glucose deprivation with one of the following treatments: addition of 97 mM mannitol, equimolar substitution of choline+ for Na+, or substitution of isethionate- for Cl-. Some retinas were resupplied with oxygen and glucose to determine the consequences of solute manipulation. Water compartments and membrane permeability were assayed using [3H]inulin and [14C]mannitol. Abnormal nuclei were counted on blind-coded slides. Both treatments prevented doubling of inulin-free water of untreated retinas after 70 min of deprivation. Neither treatment preserved morphology or prevented massive swelling after restoration of oxygen and glucose and gradual removal of the treatment. Mannitol leaked into deprived cells only, even when swelling was prevented. We conclude that prophylaxis of cell swelling during metabolic energy deprivation prevents neither cell leakiness nor cell death.

Aerobiosis↗
PubMed1984

Interleukin 3: A differentiation and growth factor for the mouse mast cell that contains chondroitin sulfate E proteoglycan.

Mouse mast cells were differentiated and grown by culturing bone marrow cells in medium containing 2 X 10(-10) M purified interleukin 3 (IL 3). The cells obtained were similar in ultrastructure, membrane antigen phenotype, proteoglycan type, and lipid products generated upon immunologic activation to mast cells differentiated in culture by WEHI-3-conditioned medium (WEHI-3-CM) and by concanavalin A (Con A) splenocyte-conditioned medium. Phenotypically, these cells expressed IgE receptors and H-2 antigens and were recognized by a monoclonal antibody (B23.1) that did not react with mouse serosal heparin-containing mast cells. The classic phenotypic markers of mouse T cells or macrophages were not detected. The mouse mast cells differentiated with IL 3 as well as those differentiated in WEHI-3-CM incorporated [35S]sulfate into a nonheparin proteoglycan of 150,000 to 200,000 m.w. Most of the 35S-labeled macromolecules were degraded by chondroitinase ABC to yield only two disaccharides, which co-chromatographed on ascending thin layer chromatography with delta Di-4S and delta Di-diSE; thus, the proteoglycan in these cells is composed of chondroitin sulfate E glycosaminoglycans. After sensitization with monoclonal IgE, washing, and antigen activation, the IL 3 differentiated cells released the preformed mediator beta-hexosaminidase and generated and released two major classes of lipid mediators. The quantities of leukotriene C4 (LTC4), leukotriene B4 (LTB4), and platelet-activating factor (PAF-acether) generated/10(6) cells were 17, 3.0, and 3.1 ng, respectively. The ratio of these three lipid mediators was similar to that obtained from mast cells differentiated in WEHI-3-CM and in Con A-conditioned medium. Thus, T cell-derived IL 3 is the component present in the conditioned media that is required for differentiation and growth of the subclass of mast cells containing chondroitin sulfate E proteoglycan, designated E-MC. The IL 3-dependent E-MC may represent the in vitro counterpart of the T-cell-dependent mucosal mast cell, suggesting in turn that the production of LTC4 and LTB4 and of PAF-acether may play a role in adaptive intestinal immunity to helminthic parasites.

Animals↗
PubMed1984

Cloned mouse mast cells derived from immunized lymph node cells and from foetal liver cells exhibit characteristics of bone marrow-derived mast cells containing chondroitin sulphate E proteoglycan.

Cloned mouse mast cells which were T cell growth-dependent were derived both from immunized lymph node and from foetal liver, and were found to be morphologically and biochemically similar to mast cells previously differentiated in vitro from mouse bone marrow (BMMC). These two T cell growth-dependent mouse mast cell clones were identical to the BMMC in their preferential synthesis of chondroitin sulphate E proteoglycan rather than heparin proteoglycan. The hydrodynamic size of the cell-associated proteoglycan from each of the three mast cell sources was 150,000-250,000 mol. wt.; and that of the covalently bound glycosaminoglycans was 13,000-25,000 mol. wt. Chondroitinase ABC digestion of the [35S]proteoglycans from both cloned mast cells, as well as the BMMC, yielded only two disaccharides which comigrated on ascending thin layer chromatography with delta Di-4S and delta Di-diSE standards, respectively. Quantification of the radioactivity in the enzyme digests revealed that one-sixth to one-half of the resulting disaccharides were disulphated, similar to that found in BMMC containing chondroitin sulphate E. When sensitized with monoclonal IgE, washed, and subsequently challenged with specific antigen, each of the two cloned mast cells generated more than 100 ng of leukotriene C4 (LTC4)/10(6) cells, but only 3-12 ng leukotriene B4 (LTB4)/10(6) cells, characteristics also observed for the BMMC. Based upon these observations, it is concluded that the cloned mast cells from lymph node and liver and the bone marrow-derived mast cell belong to a distinct subclass of mast cells. These mast cells have been designated E-mast cells (E-MC) in order to distinguish them from heparin-containing mast cells (H-MC).

Animals↗
PubMed1983

Laminin in glomerular basement membranes of aminonucleoside nephrotic rats. Increased proteinuria induced by antilaminin immunoglobulin G.

The amount and distribution of the glycoprotein laminin was investigated in the glomerular basement membranes (GBM) of rats made nephrotic by 10 daily subcutaneous injections of the aminonucleoside of puromycin (AMN). Affinity-purified, sheep antilaminin immunoglobulin G (S alpha L) was injected intravenously into AMN rats, and kidney-bound S alpha L was compared with normals. Photometric measurements of glomerular-bound S alpha L showed that intravenous injections of 1.5 mg of S alpha L saturated laminin in normal glomeruli. The same amount of glomerular S alpha L was present in 10-day AMN nephrotic rats after injection of a saturating dose. In nephrotic rats, approximately 4.4% of a dose of 0.1 to 0.9 mg of 125I-S alpha L bound in the kidneys as compared with approximately 3.8% in normals. By immunofluorescence microscopy, S alpha L bound in a linear pattern to the GBM in nephrotic and normal rats and remained similarly bound throughout all stages of nephrosis. Conjugates of S alpha L and horseradish peroxidase (S alpha L-HRP) injected into normal and 10-day nephrotic rats bound to all three layers of the GBM, to fibrillar structures within the laminae rarae, and to the plasma membranes of epithelial cells below the slit diaphragms. In nephrotic rats, S alpha L-HRP was also bound to the epithelial plasma membrane where it had detached from the GBM. Rats given S alpha L before the induction of AMN nephrosis (S alpha L-AMN) developed significantly greater proteinuria on day 10 than rats given AMN alone (372 versus 274 mg/24 hours, p less than 0.05) and on day 12 (603 versus 453 mg/24 hours, p less than 0.05). In addition, there was greater epithelial detachment from the GBM in S alpha L-AMN rats than simple AMN rats. We conclude that (a) large amounts of laminin are neither lost nor redistributed during AMN nephrosis, (b) laminin is present as fibrils within the GBM as well as on the epithelial plasma membrane adjoining the GBM, and (c) GBM-bound S alpha L promotes proteinuria during late stages of AMN nephrosis, possibly by enhancing epithelial detachment.

Animals↗