Biomedical subjects

H J Gabius

Publications and source records attributed to H J Gabius.

At least 145 records · Page 8Linked to original sources

PubMed1996

Carcinoid tumors of the lung: immuno- and ligandohistochemistry, analysis of integrated optical density, syntactic structure analysis, clinical data, and prognosis of patients treated surgically.

BACKGROUND: Analysis of potentially prognostic relevant factors of carcinoid tumors of the lung. METHODS: Clinical features, tumor size, and features derived from immuno- and ligandohistochemistry, cytometry and histometry, and survival have been analyzed in 82 potentially curatively resected carcinoid tumors of the lung. RESULTS: Patients with typical carcinoid tumors had a longer history of symptoms (13 vs. 8 months), fewer smoked (30% vs. 80%), and developed less frequently lymph node metastases (20% vs. 65%) compared to patients with atypical carcinoids. Statistically significant differences between both cell types have been observed in cytometric and histometric features, and binding of Lewis A trisaccharide (Lea). Prognosis is associated with the cell type, presence of lymph node metastases and heparin-binding lectin (HBL), certain cytometric and structural features, and binding of macrophage migration inhibitory factor (MIF) and beta-N-acetyl-D-galactosamine (beta-GalNAc). CONCLUSIONS: Complete lymph node dissection is necessary, data of cytometry, histometry, and ligandohistochemistry might eventually predict the course of the disease.

Adolescent↗

PubMed1996

Effect of phagocytosis of pHEMA particles and of heat-killed Candida albicans on expression of carbohydrate-binding sites such as endogenous lectins in phagocytes.

Phagocytosis of particles is an integral part of the defence system. Besides clearing the environment of foreign material this mechanism may affect the expression of physiologically relevant epitopes such as carbohydrate-binding sites, e.g. lectins. To determine the effect of particle design on the expression of such determinants in human monocytes and peritoneal macrophages either in suspension or after adherence, the binding of labelled (neo)glycoproteins was comparatively studied after exposure to poly (2-hydroxyethyl methacrylate) particles and heat-inactivated Candida albicans cells. A stimulatory effect of phagocytosis on extent of expression of binding sites for (neo)glycoproteins in phagocytic cells was observed. The levels of responsiveness varied according to the type of particle, adherence of the cell adding a further regulatory parameter. These results support the notion of a potential influence of the chemical structure and/or the form of an engulfed particle on phagocyte differentiation.

Binding Sites↗

PubMed1996

Binding capacities of two immunomodulatory lectins, carrier-immobilized glycoligands and steroid hormones in lung cancer and the concentration of nitrite/nitrate in pleural effusions.

Combined analysis of the binding properties of inflammatory and tumor cells in pleural effusion, and tumor imprints for various carrier-immobilized types of ligands and lectins, and of a biochemical feature of the effusions is performed to extend the characterization of these cells and their activity. In detail, the binding of Viscum album agglutinin (VAA), Urtica dioica agglutinin (UDA), and of carrier-immobilized N-acetyl-D-glucosamine (GlcNAc), lysoganglioside GM1, estradiol, progesterone, testosterone, and hydrocortisone to native specimens consisting of 46 tumor imprints from surgically treated patients with lung cancer and 74 smears of pleural effusion (PE) cells from cancer or non-cancer patients was studied using fluorescence microscopy with Texas red-labeled streptavidin. Among the tested ligands, VAA was found to provide the most effective staining of cells (60-78.1% of positive cases). When compared with inflammatory cells from PE, cancer cells were seen to bind more frequently only two ligands, namely UDA and estradiol. Significant (P < 0.001) difference between patients with bronchial carcinoma and non-cancer patients were found, when the content of NO2-/NO3- in PE fluids was measured. Whereas the level of NO2-/NO3- in PE of non-cancer patients was 12.6 +/- 10.7 microM (n = 12), it was 37.7 +/- 19.4 microM (n = 14) in cancer patients without pleural metastases and 37.5 +/- 16.0 microM (n = 26) in patients with pleural metastases. The level of NO2-/NO3- in PE appeared to correlate with extent of staining with GM1 and GlcNAc: in non-cancer patient groups it was significantly higher (P = 0.032) for negative subjects than those binding the ligand GlcNAc, whereas in the patient group with adenocarcinoma it was significantly lower (P = 0.032) for patients without binding capacities for GlcNAc and GM1. The results obtained suggest that the combined analysis of increased levels of NO2-/NO3- in PE and of glycohistochemical properties of cancer and inflammatory cells may be useful in exploring the interrelationship of functionally important cellular characteristics.

Acetylglucosamine↗

PubMed1996

Influence of intraperitoneal injection of three types of hydrogel beads on expression of carbohydrate-binding sites in spleen macrophages.

The influence of the chemical structure of polymer implants on selected characteristics of macrophages was studied to improve our understanding of the mechanisms of non-self recognition of synthetic materials. Three types of polymers differing in net charge were prepared to compare in vivo responses. Beads from preparation of poly(2-hydroxyethyl methacrylate), a copolymer of 2-hydroxyethyl methacrylate with sodium methacrylate, and a copolymer of 2-hydroxyethyl methacrylate and dimethylaminoethyl methacrylate were injected intraperitoneally into rats and harvested 48 h later. The effects of these polymers on the presence of inflammatory cells in the peritoneal exudate, on the adhesion of macrophages to individual batches of the different types of beads and on distinct molecular aspects of macrophages in the red pulp of spleen were evaluated. Beads from both types of copolymer caused an elevation in the number of macrophages in the exudate, in contrast to the situation in rats treated with poly(2-hydroxyethyl methacrylate) beads and physiological saline solution as control. The molecular design of the implant had no significant influence on the extent of macrophage adhesion to beads or on the expression of selected carbohydrate-binding sites. Since important cellular functions such as cell adhesion and glycoprotein routing depend on the sugar part of glycoconjugates, labelled neoglycoproteins were employed to analyse this aspect of macrophages in the tested animals. The beads of the copolymer of 2-hydroxyethyl methacrylate with dimethylaminoethyl methacrylate clearly led to an elevation of the expression of specific binding sites for beta-galactoside-terminating structures which are presented by asialofetuin, for mannose, fucose, sialic acid and N-acetylgalactosamine, which had been used as the ligand parts of biotinylated neoglycoproteins, in spleen macrophages whereas the levels of sites which recognize mannose-6-phosphate were unaffected. Expression of sites with specificity to N-acetylglucosamine was lessened. The effect of beads from the copolymer of 2-hydroxyethyl methacrylate with sodium methacrylate on the measured glycobiological features in the splenic macrophages was only negligible. These results suggest the possibility of systemic effects of implanted polymers on the distinct recognitive functions of macrophages.

Animals↗

PubMed1996

Effects of metabolic inhibitors and lectins on the menadione-dependent generation of H2O2 by rat thymocytes.

The capacity of oxidative metabolism and its regulation is an important factor in disease control. Using scopoletin as a fluorescent substrate of peroxidase the extent of menadione-dependent production of H2O2 by rat thymocytes was determined. The reaction was inhibited by 2,4-dinitrophenol, papaverine, nordihydroguaiaretic acid and iodacetamide. The membrane-penetrating SH reagent N-ethylmaleimide primed the reaction, probably due to an inhibition of glutathione peroxidase. To delineate an influence of cell-surface protein-carbohydrate interactions by exogenous lectins, the impact of cell binding was analyzed for several plant lectins, namely concanavalin A, phytohemagglutinin, the lectins from Triticum vulgaris and from Sambucus nigra. Except for the alpha-NeuNAc(2-6)gal/galNAc-binding agglutinin, the other three plant proteins with specificities to different parts of N-linked oligosaccharides primed the reaction. This activity of lectins did not coincide with their ability to aggregate cells. The given results indicate that biosignaling pathways triggered by lectins are involved in the regulation of the intracellular reduction of menadione.

2,4-Dinitrophenol↗

PubMed1996

Molecular dynamics-derived conformation and intramolecular interaction analysis of the N-acetyl-9-O-acetylneuraminic acid-containing ganglioside GD1a and NMR-based analysis of its binding to a human polyclonal immunoglobulin G fraction with selectivity for O-acetylated sialic acids.

The influence of 9-O-acetylation of GD1a, yielding GD1a (eNeu5,9Ac2) with a 9-O-acetylated sialic acid moiety linked to the outer galactose residue, on the spatial extension and mobility of the carbohydrate chain and on recognition by a natural human antibody is analysed. To study a potential impact of the O-acetyl group on the overall conformation of the carbohydrate chain, molecular dynamics (MD) simulations of oligosaccharide chain fragments of increasing length starting from the non-reducing end have been carried out for the first time in this study. They revealed a considerable loss in chain flexibility after addition of the internal N-acetylneuraminic acid onto the chain. Besides MD calculations with different dielectric constants, the conformational behaviour of the complete oligosaccharide chain of the 9-O-acetylated GD1a ganglioside was simulated in the solvents water and dimethyl sulfoxide. These solvents were also used in NMR measurements. The results of this study indicate that 9-O-acetylation at the terminal sialic acid does not influence the overall conformation of the ganglioside. An extended interaction analysis of energetically minimized conformations of GD1a (eNeu5,9Ac2) and GD1a, obtained during molecular dynamics simulations, allowed assessment of the influence of the different parts of the saccharide chains on spatial flexibility. Noteworthy energetic interactions, most interestingly between the 9-O-acetyl group and the pyranose ring of N-acetylgalactosamine, were ascertained by the calculations. However, the strength of this interaction does not force the ganglioside into a conformation, where the 9-O-acetyl group is no longer accessible. Binding of GD1a (eNeu5,9Ac2) to proteins, which are specific for 9-O-acetylated sialic acids, should thus at least partially be mediated by the presence of this group. To experimentally prove this assumption, a NMR study of 9-O-acetylated GD1a in the presence of an affinity-purified polyclonal IgG fraction from human serum with preferential binding to 9-O-acetylated sialic acid was performed. The almost complete disappearance of the intensity of the 9-O-acetyl methyl signal of the GD1a (eNeu5,9Ac2) clearly indicates that the assumed interaction of the 9-O-acetyl group with the human protein takes place.

Acetylation↗

PubMed1996

Comparative cross-linking activities of lactose-specific plant and animal lectins and a natural lactose-binding immunoglobulin G fraction from human serum with asialofetuin.

Plant and animal lectins bind and cross-link certain multiantennary oligosaccharides, glycopeptides, and glycoproteins. This can lead to the formation of homogeneous cross-linked complexes, which may differ in their stoichiometry depending on the nature of the sugar receptor involved. As a precisely defined ligand, we have employed bovine asialofetuin (ASF), a glycoprotein that possesses three asparagine-linked triantennary complex carbohydrate chains with terminal LacNAc residues. In the present study, we have compared the carbohydrate cross-linking properties of two Lac-specific plant lectins, an animal lectin and a naturally occurring Lac-binding polyclonal immunoglobulin G subfraction from human serum with the ligand. Quantitative precipitation studies of the Lac-specific plant lectins, Viscum album agglutinin and Ricinus communis agglutinin, and the Lac-specific 16 kDa dimeric galectin from chicken liver demonstrate that these lectins form specific, stoichiometric cross-linked complexes with ASF. At low concentrations of ASF, 1:9 ASF/lectin (monomer) complexes formed with both plant lectins and the chicken lectin. With increasing concentrations of ASF, 1:3 ASF/lectin (monomer) complexes formed with the lectins irrespective of their source or size. The naturally occurring polyclonal antibodies, however, revealed a different cross-linking behavior. They show the formation of 1:3 ASF/antibody (per Fab moiety) cross-linked complexes at all concentrations of ASF. These studies demonstrate that Lac-specific plant and animal lectins as well as the Lac-binding immunoglobulin subfraction from specific stoichiometric cross-linked complexes with ASF. These results are discussed in terms of the structure-function properties of multivalent lectins and antibodies.

Animals↗

PubMed1996

Sialic acids structure-analysis-metabolism-occurrence-recognition.

Sialic acids are commonly positioned at non-reducing termini of complex carbohydrates. Steady refinements of analytical techniques have enabled detailed mapping of the complexity of sialic acids, unravelling a number of possibilities for substitutions. These developments have aided the description of the required enzymatic activities. In view of the physiological significance of this intriguing extent of variability of one sugar unit, the assumption that distinct types of sialic acids can serve as ligands in recognitive interactions is gaining support. It is reinforced by the discovery of several classes of mammalian lectins that bind sialo-glycoconjugates. Notably, an often encountered modification of sialic acids, namely O-acetylation, can be considered as a modulatory signal in recognition, either serving as contact point or masking a ligand structure. The increased knowledge of the physiological roles of sialic acids, for example in selectin-mediated leukocyte recruitment to sites of inflammation or in influenza virus propagation, even points to clinical applications. This perspective has led the field from the inherently descriptive beginning to technically sophisticated attempts for deliberate drug design.

Animals↗

PubMed1996

[The mistletoe myth--claims, reality and provable perspectives].

Intuition guided R. Steiner to refer to mistletoe as the future remedy for cancer. He proposed that its spiritual qualities support re-establishment of the harmonious integration of the alleged four different entities of human existence in a patient. The assumption of potency without chemical basis is derived from the dogmatic system of anthroposophic reasoning. It explains the evidently similar claim of clinical efficiency for the proprietary mistletoe extracts despite the lack of information on the actual contents of the complex mixtures and despite the conspicuous diversity of methods of manufacture for these products. Thorough scientific analysis of the published clinical experience does not justify this claim. Due to the increasing reference to defined substances in advertisements for commercial extracts, they should no longer avoid rigorous testing according to common quality standards. Interdisciplinary research efforts on the immunomodulatory galactoside-binding lectin illustrate how to yield a clinically testable substance from an ill-defined extract, thereby providing a notable example for rational investigation of unconventional treatment modalities.

Adjuvants, Immunologic↗

PubMed1996

Cytometry with DAPI-stained tumor imprints. A reliable tool for improved intraoperative analysis of lung neoplasms.

OBJECTIVE: To determine the feasibility and potential use of application of 4'-6-diamidino-2-phenylindole dihydrochloride-stained imprints taken from primary lung carcinomas for intraoperative frozen section service. STUDY DESIGN: Tumor imprints taken from intraoperative sections were stained with the fluorochrome DAPI, and the integrated nuclear fluorescence (INF) was measured using an automated image analyzing system. The 40 cases studied comprised all major lung carcinoma types as well as intrapulmonary metastases. The measured features included percentage of tumor cells > 3C and > 5C, S-phase-related tumor cell fraction, 2 CV standard deviation and INF entropy. RESULTS: Test series with imprints of tumor-free lymph nodes revealed a minimum staining time of five minutes at 40 degrees C that led to reproducible staining intensities and size of the nuclear area. No significant differences were apparent in comparison to concomitantly performed measurements of the integrated optical density of "classical" Feulgen-stained specimens for each case. The fluorometric results could be submitted to the surgeon with 15-18 minutes (20 maximum). CONCLUSION: The data indicate that cytometric DNA measurements can be performed intraoperatively with reliable quality within time schedules acceptable for intraoperative surgical strategies.

DNA, Neoplasm↗

PubMed1996

Modulation of the glycoconjugate expression in the tracheo-bronchial epithelium during sustained hypovitaminosis A.

The aim of this study was to determine the influence of sustained marginal vitamin A deficiency on the morphology of glycoconjugate expression in the tracheobronchial epithelium of guinea pigs. The distribution of oligosaccharide chains was investigated by applying a panel of 24 lectins. Glycosaminoglycans were detected by histochemical techniques. Number as well as morphology of ciliated cells showed no significant alterations in hypovitaminosis A. In contrast, the quantity of goblet cells was constantly decreased. A considerable reduction of secretory granules was also observed in these cells. Cytomembranes of ciliated cells (especially in the area of ciliar extensions) showed constant alterations in the patterns of lectin binding in vitamin A-depleted guinea pigs. Our results demonstrate a significant augmentation of accessibility of fucosyl molecules in proximal domains of glycoconjugates of ciliary membranes, whereas the presence of mannose structures seemed unchanged. In distal bronchioli, terminal N-acetylgalactosamine molecules were expressed. During marginal vitamin A deficiency, ciliary cells were specially labelled by GSA I(B), indicating presentation of terminal galactose molecules in alpha-position. Additionally, the cytoplasm of epithelial cells demonstrated enhanced concentrations of polyantennary oligosaccharide core structures. Staining of epithelial cells by VVA was restricted to control specimens. Abundance of N-acetylglucosamine residues on the non-reducing terminus of oligosaccharides was significantly enhanced in the connective tissue of depleted animals as demonstrated by the binding patterns of GSA II. We suggest that altered oligosaccharide patterns may contribute to enhanced predisposition to tracheobronchial infection in marginal vitamin A deficiency.

Animals↗

PubMed1996

Application of attributed graphs in diagnostic pathology.

OBJECTIVE: To compute attributed graphs based upon calculation of the minimum spanning tree (MST) for various applications in diagnostic lung pathology. STUDY DESIGN: The study design included assistance in histologic diagnosis, confirmation of the diagnosis in single cases, measurement of texture alterations after induction chemotherapy, estimation of prognosis of operated-on lung cancer patients and analysis of lung cancer cells in association with differentiation markers. The histologic slides were Feulgen stained, and features of the integrated optical density (IOD) were associated with the nodes of the MST. The same procedure was applied to immunohistochemically and ligand histochemically stained slides by calculation of the mean staining intensity of the cytoplasm of tumor cells. A measure for structural entropy was introduced by computing the relative differences in distance and IOD between neighboring tumor cells in a 1/r2 field of force. In addition, the current of entropy was computed. RESULTS: Structural entropy reflects alterations in regular textures; the current of entropy is an especially good prognostic parameter in lung cancer. In immunochemistry and ligand histochemistry, construction of the attributed MST permits detailed insight into locally different staining behavior of tumor cells and immunocompetent cells. CONCLUSION: Attributed graphs contain important information that can be used for the estimation of survival or for confirmation of diagnostic entities, such as tumor cell types.

Adolescent↗

PubMed1996

Histochemical study of expression of lectin-reactive carbohydrate epitopes and glycoligand-binding sites in normal human appendix vermiformis, colonic mucosa, acute appendicitis and colonic adenoma.

In a glycohistochemical analysis of human appendix vermiformis we report the assessment of lectin binding in cells of the Gut Associated Lymphoid Tissue of normal samples and in acute appendicitis using a panel of plant, invertebrate and mammalian lectins with specificity for alpha-L-Fuc (UEA-I), alpha-D-Gluc and alpha-D-Man (Con A), alpha-D-GalNAc (DBA), GalNAc (SBA, HPA), beta-Gal (RCA-I, 14 kDa = galectin-1) and alpha-, beta-Gal (VAA). Moreover, we initiate the study of expression of carbohydrate-binding sites in this tissue and in colonic mucosa, employing several types of carrier-immobilized carbohydrate ligands as suitable probes for this purpose. Within the three populations of macrophages intra-/subepithelial macrophages of the dome region, the lamina propria of the intercryptal region and the follicle-associated epithelium were apparently reactive with most of the lectins and also with mannose and fucose residues of the tested neoglycoproteins. Distinguishing features of germinal center macrophages in relation to intra-/subepithelial phagocytes were the lack of binding of UEA-I and DBA. In comparison to all other types of phagocytes, macrophages of the T-region displayed a rather restricted binding capacity only to Con A and RCA-I. Labeling of macrophages with SBA, HPA and VAA in this location was only rarely found. With respect to dendritic cells no consistently positive reaction was seen for follicular cells, whereas interdigitating cells of the T-region bound Con A, HPA and RCA-I, and, less frequently, SBA. Lymphocytes in all anatomical subsites of the Gut Associated Lymphoid Tissue, centrocytes, centroblasts and plasma cells had binding sites for Con A and RCA-I in common. Notably, a small number of lymphocytes mostly in the T-region but also in B-cell-rich areas expressed intranuclear binding sites for fucose and mannose residues. Intraepithelial lymphocytes and lymphatic cells of the T-region differed from lymphocytes in other regions by a more frequent expression of VAA-binding sites. The epithelium of appendix vermiformis and colonic mucosa not only presents lectin binding sites, but also has the capacity to bind carbohydrate structures, as shown by labeled glycoligand-exposing neoglycoproteins. In normal mucosa the extent of binding appeared to be associated with maturation of cells, the surface epithelium showing the most intense staining reaction. This pattern is not detectable in colonic adenoma which reveal increased intensity, when compared to normal mucosa. In contrast to development of hyperplasia, acute inflammation in appendicitis caused no detectable changes of neoglycoprotein binding. Taking our previous assessment on lectin binding in appendicitis into account, we conclude that glycosylation of goblet cell mucus, but not the capacity to bind certain sugar epitopes responds to inflammatory processes, whereas tumorigenesis of colonic adenoma can also affect the binding of neoglycoproteins.

Adenoma↗

PubMed1996

Histochemical detection of expression of binding sites for labelled hyaluronic acid and carrier-immobilized synthetic (histo-blood group trisaccharides) or biochemically purified (ganglioside GM1) glycoligands in nasal polyps and other human lesions including neoplasms.

This study is intended to demonstrate the versatility and feasibility of custom-made oligosaccharide-exposing neoglycoconjugates including histo-blood group epitopes in various human lesions, including nasal polyps. The binding of the biotinylated probes was determined on formalin-fixed paraffin-embedded sections from archive materials. The general aspects of our results may be interpreted as follows: the neoglycoconjugates used here can readily detect differences in the ability of cells to bind glycan residues in tissue sections, thereby enabling the extent of the binding capacity of various types of human lesions to be compared. Furthermore, the reactivity to glycan may reflect characteristics of the cells and their environment. The investigation into pathological disorders with respect to the binding capacity of these carrier-immobilized mono- or oligosaccharide structures derived from custom-made synthesis or biochemical purification is based on the prospect of translating progress in this field into the establishment of potentially beneficial procedures for medical diagnosis and pathological classification.

Adenocarcinoma↗

PubMed1995

Effect of enzymatic desialylation of human serum amyloid P component on surface exposure of laser photo CIDNP (chemically induced dynamic nuclear polarization)--reactive histidine, tryptophan and tyrosine residues.

The human pentraxin serum amyloid P component (SAP) exhibits no microheterogeneity in its complex di-antennary glycan. To elucidate whether the removal of sialic acids from this glycoprotein might affect the accessibility of certain amino acid residues of the protein we employed the laser photo CIDNP approach as a sensitive tool. The CIDNP effect is created by the interaction of a photoexcited dye with reactive amino acids and results in enhanced absorption- or emission-signals which can be observed for the three aromatic amino acids histidine, tryptophan, and tyrosine if they are accessible to the dye. Therefore, this technique can be applied to explore surface exposure of these residues. The respective spectra of SAP and enzymatically desialylated SAP were determined. Six tryptophan/histidine signals and one tyrosine signal are present in the aromatic part of the CIDNP difference spectrum of SAP. The corresponding spectrum of desialylated SAP shows remarkable alterations. The chemical shift of one tryptophan/histidine-characteristic signal is decreased by 0.1 ppm. One tryptophan/histidine signal disappeared and a new one was formed in the CIDNP difference spectrum of desialylated SAP, while the other signals were unaffected. The tyrosine signal has a clearly enhanced intensity in desialylated SAP. Therefore, the removal of sialic acid moieties from the single N-glycan of each monomer apparently affects surface presentation of distinct CIDNP-reactive amino acids of SAP.

Amino Acids↗

PubMed1995

Carcinoembryonic antigen and other glycoconjugates act as ligands for galectin-3 in human colon carcinoma cells.

Galectin-1 and galectin-3, galactoside-binding lectins with molecular weights of M(r) 14,500 and 31,000, respectively, are expressed in normal and malignant cells and have been implicated in regulation of cell growth, adhesion, and metastasis. We analyzed the expression of galectins in 21 cultured human colon carcinoma cell lines by immunoblotting. Galectin-1 was detected in only 7, whereas galectin-3 was found in 20 of the cell lines. KM12 cells, which express only galectin-3, were used to isolate this lectin by affinity chromatography, and the purified lectin was used to identify complementary glycoconjugates by blotting. Galectin-3 was shown to bind to human laminin, carcinoembryonic antigen, and lysosome-associated membrane glycoproteins, which are involved in cell adhesion. Galectin-3 was localized on the KM12 cell surface and colocalized with carcinoembryonic antigen. Several endogenous glycoproteins and cell surface proteins of molecular weights in the range M(r) 58,000 to > 200,000, including carcinoembryonic antigen and lysosome-associated membrane glycoproteins, were identified as galectin-3 ligands by coimmunoprecipitation with and affinity chromatography on immobilized galectin-3. These data demonstrate that galectin-3 interacts with several adhesion molecules and suggest that this lectin may have a role in human colon carcinoma cell adhesion.

Antigens, CD↗

PubMed1995

Differential response of the epidermal growth factor receptor tyrosine kinase activity to several plant and mammalian lectins.

Biosignalling via lectins may involve modulation of protein kinase activities. This aspect of the biological action of mammalian and plant lectins has been investigated for their effect on the activity of the isolated epidermal growth factor receptor (EGFR). The constitutive tyrosine kinase activity of the epidermal growth factor receptor from rat liver, isolated by calmodulin-affinity chromatography, was activated by concanvalin A (ConA), and wheat germ agglutinin (WGA) to a similar extent as the measured enhancement induced by EGF. In contrast, two mannose-specific lectins, the mannan-binding protein (MBP) and serum amyloid P component (SAP), isolated from human serum, have inhibitory effects, both in the absence and presence of EGF. The differential effects of these lectins were tested using as phosphorylatable substrates a co-polymer of glutamic acid-tyrosine, as well as calmodulin. However, two galactoside-specific lectins, the laminin-binding beta-galactoside-binding 14 kDa lectin, isolated from bovine heart (14K-BHL), and the alpha/beta-galactoside-binding lectin, isolated from mistletoe (Viscum album L.) leaves (VAA), do not inhibit the EGFR tyrosine kinase activity. The sugar dependence of the lectin-mediated action was studied by inhibition assays. Mannose and a mannose-containing neoglycoprotein prevent the activating effect of ConA, and N-acetyl-D-glucosamine partially prevents the activation produced by WGA. However, mannose and mannose-containing neoglycoprotein were ineffective to reduce the inhibitory effect of MBP or SAP.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylglucosamine↗

PubMed1995

Differential binding of two chicken beta-galactoside-specific lectins to homologous lymphocyte subpopulations and evidence for inhibitor activity of the dimeric lectin on stimulated T cells.

Plant lectins can be potent modulators of vertebrate immune functions. Biochemical characterization of lectins from animal tissues enables the determination of whether these endogenous activities display a comparable immunological potency. Focusing on chicken beta-galactoside-binding lectins, the monomeric intestinal (CL-14) and the dimeric liver lectin (CL-16) were purified and the lack of cross-contamination was ascertained. In very close agreement with the molecular masses of 14,974 and 14,976 calculated on the basis of the available sequence data (Y. Sakakura et al., J. Biol. Chem. 265, 21573-21579, 1990), electrospray mass spectrometric analysis yielded values of 14,969 (CL-14) and 14,972 (CL-16), the reasons for the deviation in gel electrophoretic behavior being unclear. Solid-phase assays with immobilized lactosylated poly-L-lysine demonstrated a comparatively lower affinity and higher extent of binding at saturation for the monomeric lectin than for the dimeric protein, whose properties were similar to those of an immunomodulatory plant lectin. Flow cytometry revealed homogeneous and strong binding of the dimeric lectin within the chicken peripheral blood lymphocyte population, whereas the monomeric lectin stained two subpopulations at different intensities. Two-color flow cytometry disclosed preferential binding of this lectin to B cells. When a B cell line was employed for determination of affinity constants and extents of binding at saturation, qualitatively comparable parameters to those for the solid-phase assays were obtained. The similar profile of lectin-binding glycoproteins in blots of cellular extracts underscored that accessibility to ligands, not qualitatively different ligand display, may explain the differences for the cell line. At up to a concentration of 10 micrograms/ml of the lectins no stimulation of [3H]thymidine incorporation was seen for blood and spleen cell populations. However, the dimeric lectin reduced stimulation of cells that were responsive to an anti-TcR2 antibody. Thus, this lectin can apparently exhibit inhibitory activity to this kind of T cell activation in vitro.

Animals↗