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H J Gabius

Publications and source records attributed to H J Gabius.

At least 271 records · Page 15Linked to original sources
PubMed1989

Lineage- and differentiation-dependent alterations in the expression of receptors for glycoconjugates (lectins) in different human hematopoietic cell lines and low grade lymphomas.

Important biological functions and cellular recognition phenomena are supposedly governed by specific sugar-protein interactions. Human hematopoietic cell lines offer an excellent model for the study of the expression of endogenous receptors for the carbohydrate part of glycoconjugates with respect to cell lineage and modulation by differentiation. Initially, a panel of fluorescent (neo)glycoproteins was successfully employed to demonstrate cytologically the actual presence of such receptors on different cell lines: the B lymphoblast line, Daudi; the T cell lymphoblastic leukemia line, P12; the multipotent leukemic line, K562 and the promyelocytic line, HL060. Biochemical analyses were performed using affinity chromatography on supports with immobilized lactose and asialofetuin (simple or complex beta-galactosides), melibiose (alpha-galactoside), fucose, N-acetyl-D-galactosamine, maltose (alpha-glucoside), the mannose-rich yeast glycoprotein, mannan, glycopeptides containing sialic acid residues and heparin. Subsequently, sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis was used to detect cell lineage-dependent changes in theses parameters. Differentiation-dependent changes in the expression of receptors with specificity to galactose, N-acetylgalactosamine, maltose and heparin were similarly uncovered upon dimethyl sulfoxide-induced differentiation of HL60 cells. Differences in this type of cellular characteristic were also apparent for lymphoma cells from patients with various histological subtypes of lowgrade lymphomas. This initial description of lineage- and differentiation-dependent differences in various human hematopoietic cell lines and in cells from patients with lowgrade lymphomas suggests that advances in the knowledge of the composition of endogenous sugar receptors (lectins) may aid in understanding aspects of the biological behavior of hematopoietic cells and their related malignancies via participation of sugar-protein (lectin) interactions.

Cell Differentiation↗
PubMed1989

Enhancement of clearance of plant lectins as radiopharmaceuticals by chemically glycosylated antilectin antibody.

The influence of chemical glycosylation on clearance of plant lectins, employed as radiopharmaceuticals, was assessed. Controlled chemical modification of the antibody to concanavalia ensiformis agglutinin did not affect its immunologic activity, but led to rapid clearance from circulation. In tumor-bearing mice, notable increases of selected tumor/non tumor ratios even after only 1 h of presence of the galactosylated antibody to the lectin in circulation were observed. Similarly, galactosylated antibody to pisum sativum agglutinin caused significant increases of tumor/non tumor ratios.

Animals↗
PubMed1989

Malignant and normally developing trophoblastic cells of human placenta display different characteristics defined by histochemical and biochemical mapping of endogenous lectins.

Trophoblastic cells with their strictly controlled propensity for invasive growth hereby bear limited resemblance to malignant cells. Therefore, detailed description of molecular characteristics of this cell type in comparison to histologically related tumor cells may aid in defining physiologically relevant differences. In view of the potential importance of selective protein-carbohydrate interactions in recognitive processes, labelled neoglycoproteins were employed to evaluate systematically the presence and distribution of sugar receptors in tumor cells of chorionepithelioma and in trophoblastic cells of an early stage of gestation. Control reactions in glycohistochemistry, involving prolonged incubation, pH variation and use of nucleotides excluded the possibility that glycosidases or glycosyltransferases govern the sugar-specific binding to the tissue sections. Pronounced differences were detected in the expression of receptors (lectins) for alpha- and beta-glucosides as well as for N-acetylgalactosamine and N-acetylglucosamine. Further differences were revealed by probing both types of tissue with lactosylated, fucosylated, xylosylated and sialylated carrier protein. Upon developmental maturation of the normal cells in placenta the extent of binding of the neoglycoproteins decreased. The glycohistochemical differences between malignant and normally developing trophoblastic cells were found to be reflected in the alteration of expression of certain endogenous lectins, as ascertained by affinity chromatography and gel electrophoretic analysis. Consequently, we assume that the transient presence of certain endogenous lectins during early stages of gestation and their differential expression in relation to tumor cells of chorionepithelioma may be relevant for the special invasive and adhesive behavior of human trophoblastic cells.

Choriocarcinoma↗
PubMed1989

Endogenous sugar receptor pattern in human glioblastomas and gangliocytomas studied by histochemical application of biotinylated (neo)glycoproteins and affinity chromatography.

Biotinylation of chemically glycosylated bovine serum albumin, yielding a panel of neoglycoproteins, and of desialylated, naturally occurring glycoproteins allowed to systematically evaluate presence and distribution of various types of endogenous sugar receptors in the sections of human glioblastomas and gangliocytomas by a routine histochemical procedure. Pronounced cytoplasmic staining with markers, carrying constituents of natural glycoconjugates, e.g. for beta-galactoside-specific receptors, contrasted with the different intensities, noticed for alpha- and beta-glucoside-specific receptors. Significant qualitative differences between the two tumor types were detected with N-acetyl-D-galactosamine- and sialic acid-carrying probes. Nuclear staining with only a part of the applied panel underscored the specificity of the protein-carbohydrate interaction. Fine structural features of the synthetic neoglycoproteins, e.g. the mode of coupling of the carbohydrate moiety to the protein, were found to exert a significant influence on their suitability as histochemical markers. On the basis of the histochemical results, exemplary biochemical analysis of certain classes of endogenous sugar receptors by affinity chromatography and subsequent gel electrophoresis, namely of beta-galactoside-, alpha-fucoside-, alpha-mannoside- and alpha-glucoside-specific proteins, revealed presence and characteristics of respective sugar receptors that can contribute to the histochemical staining. Similar extent of histochemical staining with the respective probes notwithstanding, the different tumor types exhibited qualitative differences in the expression of individual endogenous sugar receptors. The combined histochemical and biochemical analysis is supposed to be of conspicuous value for biological and clinical investigations on endogenous sugar receptors.

Chromatography, Affinity↗
PubMed1989

Histopathologic evaluation of application of labeled neoglycoproteins in primary bronchus carcinoma.

Neoglycoproteins are readily available conjugates of a histochemically inert carrier protein and histochemically crucial carbohydrate moieties which are covalently attached to the carrier protein by chemical synthesis. Biotinylation renders these conjugates detectable in formalin-fixed, paraffin-embedded tissue sections of human lung cancer by standard staining protocols, thereby localizing endogenous receptors for carbohydrate moieties. Examination of 30 cases of main types of human lung cancer revealed the presence of alpha-fucosyl-, alpha-mannosyl-, and alpha-glucosyl-specific receptors in adenocarcinomas or epidermoid carcinomas with high positivity rates. The extent of the expression of receptors for alpha- and beta-galactosides appeared to be comparatively lower. Within the standard protocol, using a concentration of the biotinylated probes of 10 micrograms/mL, this panel of probes consistently failed to detect endogenous sugar receptors in ten cases of small cell anaplastic carcinoma of the lung. Whereas none of the sections from the tumor cases bound the sulfated fucan fucoidan, the accompanying inflammatory cells, especially the granulocytes, expressed receptors for the sulfated fucan. Pronounced labeling for macrophages was observed for the alpha-galactoside-specific probe, whereas no binding to inflammatory cells and pneumocytes was detectable for the beta-galactoside-specific probe. The results indicate that expression of endogenous receptors for neoglycoproteins may be useful in discriminating between small cell and non-small cell lung carcinoma and carcinomatous cells from accompanying inflammatory cells.

Carbohydrate Metabolism↗
PubMed1989

Is the profile of binding of a panel of neoglycoproteins useful as a diagnostic marker in human lung cancer?

Histomorphological evaluation of the profile of carbohydrate-binding proteins with specificities to alpha- and beta-glucosides (maltose, cellobiose), alpha-L-fucosides (L-fucose), alpha-D-mannosides (D-mannose), a N-acetylated sugar, present in natural glycoconjugates (N-acetyl-D-glucosamine), two negatively charged sugars (glucuronic acid and sialic acid), rhamnose (6-deoxy-L-mannose) or to heparin has been assessed. 133 cases of formalin fixed, paraffin embedded human lung carcinoma specimens (45 cases of epidermoid carcinoma, 24 cases of small cell anaplastic carcinoma, 48 cases of adenocarcinoma, and 16 cases of large cell anaplastic carcinoma) as well as 14 cases with epithelial or biphasic mesothelioma and 7 cases of pleuritis carcinomatosa were incubated with biotinylated, suitably modified neoglycoproteins or heparin for detecting the corresponding endogenous sugar receptors (lectins). Histochemical reaction was demonstrated by use of the avidin-biotin-method. Significant differences were obtained between the different markers used for each cell type as well as between the different cell types. For separating small cell anaplastic carcinoma from non small cell anaplastic carcinoma the markers carrying the histochemically crucial maltose, fucose, and mannose residues are useful, showing a positive reaction to only 4-8% of the small cell anaplastic carcinoma cases compared to 60-80% of the non small cell carcinoma cases. For separation of mesothelioma from pleuritis carcinomatosa a carrier, modified by attachment of N-acetylglucosamine moieties is the most useful marker being negative in all cases of the analyzed mesothelioma cases and being positive in 62-80% of the adenocarcinoma cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Biomarkers, Tumor↗
PubMed1989

[Endogenous lectins in tumors and their possible importance in the diagnosis and therapy of cancers].

The oncological application of knowledge of biological recognition processes offers opportunities to favorably translate results from basic science into clinical procedures within tumor diagnosis and therapy. Based on the attractive assumption that biological information can be stored in a glycobiological code within the sugar chains of cellular glycoconjugates, the specific interaction of proteins and carbohydrates warrants special attention. Lectins form a particular class of carbohydrate-binding proteins, separated from sugar-specific enzymes as well as antibodies. They are expressed by normal cells, but also, remarkably, by tumor cells. The pattern of lectin expression can be recorded by routine histopathological procedures using labelled carriers that have been chemically modified by defined coupling of the histochemically crucial carbohydrate moieties. The tumor-associated differences, documented by application of these neoglycoproteins, serve as guidelines for consequently following biochemical studies and for lectin-mediated drug targeting. Increased selectivity of drug delivery by using tailor-made neoglycoproteins as transport vehicles may result from specifically aiming at lectins as cellular targets for binding of the 'Trojan horse'. Besides these aspects of tumor lectinology the assumption of the noteworthy significance of protein-carbohydrate recognition may also contribute to progress in a challenging as well as complex problem in tumor research, the formation of metastases. Joint approaches with chemical, biochemical, cell biological, histochemical and oncological techniques will allow to critically assess the value of tumor lectinology for oncology.

Animals↗
PubMed1989

Expression of endogenous receptors for neoglycoproteins, especially lectins, that allow fiber typing on formaldehyde-fixed, paraffin-embedded muscle biopsy specimens. A glycohistochemical, immunohistochemical, and glycobiochemical study.

A panel of biotinylated (neo)glycoproteins was used for specific detection of endogenous sugar receptors, especially lectins, in formaldehyde-fixed, paraffin-embedded muscle biopsy specimens from human deltoid, quadriceps, and biceps muscles, tibial and quadriceps muscles of rat, and bovine masseter muscle. The glycohistochemical probes used consisted of conjugates of a labeled, histochemically inert carrier protein and various covalently linked, histochemically crucial sugar moieties. Specific binding of alpha-L-fucoside, beta-D-galactoside, beta-D-xyloside, and alpha-D-mannoside to muscle sections was detected, showing no species-specific differences. The presence of receptors for the N-acetylated sugars in natural glycoconjugates, and for sugars with a phosphate group, i.e., mannose-6-phosphate and galactose-6-phosphate, was demonstrated glycohistochemically. However, these binding specificities revealed species-specific differences, e.g., the absence of N-acetyl-D-galactosamine-specific receptors or galactose-6-phosphate-specific receptors in rat muscle. Other charged sugars included glucuronic acid and sialic acid, which bound only to ox and rat muscle or failed to reveal their respective receptors in all types of muscle investigated. This different extent of staining with anionic probes served as a further control to ascertain carbohydrate binding specificity. Positive glycohistochemical reaction developed within sarcomeres only at the level of A-bands. Granular staining was observed in the sarcoplasm among the myofibrils and also in the subsarcolemmal regions. Differences in expression of glycohistochemically detectable sugar receptors were noted between type 1, type 2A, and type 2B fibers. The molecular properties of one type of glycohistochemically detectable sugar receptor were inferred both immunohistochemically and biochemically. An antiserum against an endogenous beta-galactoside-specific lectin from muscle tissue localized this lectin within sections consistently similar to (neo)glycoproteins, detecting beta-galactoside-specific receptor(s). This similarity of binding patterns strongly supports the assumption that (neo)glycoproteins with beta-galactoside termini indeed bind to the respective endogenous lectin. The lectin-specific antiserum enabled us to ascertain that glycohistochemical fiber typing corresponds to enzyme histochemical typing. Moreover, biochemical purification using affinity chromatography and subsequent affinity elution revealed only the immunohistochemically detectable beta-galactoside-specific lectin. Consequently, use of a panel of neoglycoproteins, when frozen sections for histochemical analysis are not available, co

Acetylation↗
PubMed1989

Are glycoconjugates and their endogenous receptors involved in the fusion of mononuclear macrophages resulting in multinucleate giant cells? Histochemical and electron microscopic determination of endogenous sugar-binding proteins (lectins) in mononuclear macrophages and multinucleate giant cells appearing in granulomatous foreign body reaction.

Protein-carbohydrate recognition has been found to play an important role in phagocytosis. Labelled (neo)glycoproteins were employed to comparatively analyze the histochemical pattern and ultrastructural localization of endogenous carbohydrate-binding proteins (lectins) of mononuclear macrophages and multinucleate giant cells involved in the granulomatous foreign body reaction. Sugar receptors having an affinity to simple alpha- and beta-galactoside-structures, to alpha-mannose residues, to N-acetylglucosamine, to N-acetylgalactosamine and to glucuronic acid, respectively, were detected in both cell types. However, alpha-fucoside- and beta-xyloside-specific receptors were present only in the mononuclear macrophages. Pronounced differences were seen with labelled, suitably modified glycoproteins, exposing different complex sugar parts with common beta-galactoside-termini. Among the population of multinucleate giant cells, a positive histochemical reaction was observed with mannose-6-phosphate-, galactose-6-phosphate- and glucuronic acid-(BSA-biotin), respectively, only in giant cells in which fusing mononuclear cells were recognizable. This transient expression indicates changes within the profile of endogenous sugar receptors in the stages from fusion to establishment of giant cells. Aside from the diffuse intracytoplasmic distribution of carbohydrate-binding proteins, a prominent accumulation of various types of glycosylated ferritin, used as a marker for electron microscopic evaluation, was ultrastructurally found in membranous subcellular structures and vesicles. This study is a basis for further investigation of the potential involvement of various sugar receptors in the process of macrophage fusion, resulting in multinucleate giant cells of foreign body type, and the process of phagocytosis.

Biopsy↗
PubMed1989

Neoglycoprotein binding distinguishes distinct zones in the epithelia of the porcine eye.

The presence and location of specific endogenous lectins in the anterior epithelia of the porcine eye were determined by histochemical staining using biotinylated glycosylated carrier molecules. Endogenous lectins with specificities for galactosides, alpha-mannosides and beta-xylosides were present in the conjunctival epithelium, but the corneal anterior epithelium appeared essentially free of histochemically demonstrable endogenous lectins. The corneal posterior epithelium contained a rich and complex set of endogenous lectins. The transitional area between conjunctival and corneal anterior epithelia at the limbus cornea contained, in striking contrast to either of the epithelia bordering it, cells expressing endogenous lectins in considerable wealth and variety.

Animals↗
PubMed1988

Endogenous sugar-binding proteins in human breast tissue and benign and malignant breast lesions.

Binding of carbohydrate moieties was detected in tissue sections of human breast by employing two types of labeled ligands: neoglycoproteins (chemically glycosylated, histochemically inert carrier protein) and desialylated naturally occurring glycoproteins. Paraffin-embedded, formalin-fixed sections from 40 benign and malignant breast lesions were examined for the presence and distribution of endogenous sugar receptors, employing a panel of 13 biotinylated neoglycoproteins, representing carbohydrates commonly found in naturally occurring glycoconjugates, and four biotinylated glycoproteins. Benign and malignant breast lesions revealed staining with mannosylated carrier neoglycoprotein in comparison to normal breast. A mixed pattern of staining localization and intensity was seen for different types of malignancy with this neoglycoprotein. Similarly, receptors for lactose and N-acetylglucosamine could only be detected within the cytoplasm for certain types of malignancy. Their nuclear localization, however, could also be seen in normal breast specimens. The extent of staining with different glycoproteins, containing different types of galactoside-terminal sugar chains, also appeared to differ between various types of breast cancer. The detection of endogenous sugar receptors by neoglycoproteins is proposed to contribute to an understanding of malignancy-associated alterations in the structure of their potential physiological ligands, the glycoconjugates. Changes in the structure and abundance of such glycoconjugates have commonly been detected with the use of plant lectins in histopathologic studies.

Breast Neoplasms↗
PubMed1988

An 8- to 10-fold enhancement in sensitivity for quantitation of proteins by modified application of colloidal gold.

We have modified the highly sensitive protein assay of C. M. Stoscheck (1987, Anal. Biochem. 160, 301-305), resulting in a further 8- to 10-fold enhancement of sensitivity. This assay, responding to protein quantities with a detection limit of 1 ng, involves the single step of addition of colloidal gold solution, as now commonly used in histochemistry and protein blotting, to the protein sample, followed by simple measurement of the change in absorbance at 590 nm within minutes. By increasing the concentration of the colloidal gold, by using gold sol that has been stabilized with 0.01% polyethylene glycol and adjusted to pH 3.8, and by adapting the assay to microtiter plates, this type of assay can be applied to reliably determine proteins in the complete nanogram range. This assay therefore compares favorably to other assay procedures in terms of rapidity, sensitivity, expense, and lack of interference by many laboratory reagents, although like the others it suffers from the drawback of differences in response of different proteins, which is inherent in dye-binding assays.

Colloids↗
PubMed1988

Biodistribution of neoglycoproteins in mice bearing solid Ehrlich tumor.

Synthetic neoglycoproteins were employed for the specific detection of their corresponding cellular sugar receptors, such as endogenous lectins, by specific protein-carbohydrate interaction. A panel of 16 radioiodinated probes with defined carbohydrate content, attached to the carrier protein bovine serum albumin, disclosed marked differences in the expression of corresponding sugar receptors on Ehrlich ascites tumor cells in vitro as an exemplary tumor model. To quantify tumor uptake in the more complex in vivo situation and to assess binding to individual organs attributable to the various types of carbohydrates we determined the biodistribution of the radiolabelled neoglycoproteins 48 h after injection into tumor-bearing mice. The individual pattern of retention of radioactivity demonstrated distinct properties of the different organs that need to be accounted for in drug-targeting and tumor-imaging studies, based on protein-carbohydrate interactions. Combined tumor-to-blood and tumor-to-muscle ratios of neoglycoprotein accumulation were well above 1 after covalent attachment of xylose or glucuronic acid, respectively, to the carrier protein. These data constitute the basis for further refinements of the carbohydrate part of suitable neoglycoproteins to allow a potentially rational application of neoglycoproteins as drug-targeting vehicles and tumor-imaging radiopharmaceuticals.

Animals↗
PubMed1988

Surface sugar binding components of bovine spermatozoa as evidence by fluorescent neoglycoproteins.

In the present study, the topographical distribution of carbohydrate binding sites on the plasma membrane of bovine epididymal spermatozoa was investigated using 15 different fluorescent neoglycoproteins and asialoglycoproteins. With mannose-bovine serum albumin (BSA)-fluoresceinthiocarbamyl (FTC), mannose-6-phosphate-BSA-FTC, lactose-BSA-FTC, maltose-BSA-FTC, asialolactoferrin-FTC and asialotransferrin-FTC a marked fluorescence was observed in the postacrosomal area. These results further substantiate the concept that carbohydrate binding sites of the spermatozoan plasma membrane and corresponding carbohydrates of the zona pellucida play a significant role in gamete interactions.

Animals↗
PubMed1988

Endogenous carbohydrate-binding proteins in oligodendrogliomas. A histochemical study.

Differentiated and anaplastic oligodendrogliomas revealed intracytoplasmic and nuclear presence of endogenous carbohydrate-binding proteins by application of labelled (neo)glycoproteins in histochemical analysis. The histochemical patterns showed differences between the differentiated and anaplastic forms of the same tumor type. Xylose-, lactose- and asialofetuin-specific carbohydrate-binding proteins could be detected in both types of tumors with the same staining intensity. However, maltose-specific carbohydrate-binding proteins were present only in the differentiated form. An inverse intensity of the histochemical reaction was observed with galactose-6-phosphate-, galactose-beta(1.3)-N-acetylglucosamine-N-acetylglucosamine- and mannose-(BSA-biotin) and fucose-(BSA-biotin) respectively, when the differentiated and anaplastic oligodendrogliomas were compared with each other. These differences document changes in the pattern of histochemically detectable carbohydrate-binding proteins, suggesting a role for endogenous carbohydrate-binding proteins in the tumor cell differentiation. These data indicate the potential usefulness of labelled (neo)glycoproteins as a new type of marker for histopathological diagnosis.

Carrier Proteins↗
PubMed1988

(Neo)glycoproteins as tools in neuropathology: histochemical patterns of the extent of expression of endogenous carbohydrate-binding receptors, like lectins, in meningiomas.

Biotinylated (neo)glycoproteins were used to specifically detect endogenous sugar receptors such as lectins in sections of formaldehydefixed, paraffin-embedded tissue from meningiomas. The histochemical methods used consisted of the application of a carrier protein and various covalently linked sugar moieties, available mainly through chemical synthesis, in an optimized standard protocol. They proved valuable in elucidating differential binding patterns within the various meningioma subtypes. alpha-Fucoside-, beta-galactoside-, alpha-mannoside- and beta-xyloside-specific carbohydrate-binding receptors were detected in all the tumor subclasses examined, although the levels of expression exhibited pronounced quantitative differences. In addition, differences in the extent of histochemical staining were observed, using a labelled carrier protein, derived from N-acetylglucosamine and mannose-6-phosphate moieties, respectively. Quantitative differences in the reaction intensity were also measured in the respective subtypes. Receptors for N-acetyl-D-galactosamine were detected only in the analplastic forms, while glucuronic acid-specific receptors were only present in the meningotheliomatous meningioma. In contrast to the other types, malignant meningiomas failed to show cytoplasmic staining with the alpha-glucoside-specific maltose-(BSA-biotin). Distinct differences in the pattern of expression of endogenous sugar receptors, evaluated by a standard protocol, provided further evidence for a possible additional subtype of meningioma, the submalignant meningioma. Our results suggest that labelled (neo)glycoproteins could be used routinely as tools for assessing the expression of endogenous sugar receptors in diagnostic neuro-oncology.

Biomarkers, Tumor↗