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Biomedical subjects

W Müller-Ruchholtz

Publications and source records attributed to W Müller-Ruchholtz.

At least 55 records · Page 3Linked to original sources
PubMed1989

Immunoelectron microscopic demonstration of antigenic sites on lymphoid cells using a human monoclonal antibody (Ha6D3).

The reactivity of a human monoclonal antibody directed against human B and T lymphocytes was tested for the first time at the ultrastructural level. The antigenic sites detected by this antibody were localized on the surface membrane of lymphocytes and, to a lesser extent, in the cytoplasm on membranes of the endoplasmic reticulum and perinuclear envelop of some centroblasts and immunoblasts. Ultrastructural demonstration of target antigen detected by human monoclonal antibodies may be important prior to therapeutic application of these antibodies.

Antibodies, Monoclonal↗
PubMed1989

Changes in strain-specific urine odors of mice due to bone marrow transplantations.

The change in strain-specific urine odors which appears after bone marrow transplantations was systematically examined in mice in order to demonstrate an influence of the hematopoietic system on urinary chemo-signals. Four mice were trained in a Y maze to discriminate two fully allogeneic mouse strains via their urine odors. Urine samples obtained from three inbred strains, from syngeneic reconstituted mice, and from allogeneic reconstituted chimeras were combined in a number of 'transfer of training' tests. The strain-specific urine odors of the recipients were changed by a fully allogeneic bone marrow transplant. Since this change could not be found in syngeneic reconstituted mice, we concluded that it was caused by the graft. Experiments partly failed to demonstrate donor- and recipient-specific components in the urine odor of the chimeras.

Animals↗
PubMed1989

New principle for large-scale preparation of purified human pancreas islets.

Because successful human islet transplantation requires large quantities of viable islets that must be separated from the highly immunogenic exocrine tissue and because handpicking is too time-consuming and laborious to be clinically relevant, a new approach for solving this problem has been established in rat models. It is based on the principle that magnetic microspheres (MMSs) coupled to lectins with binding specificity for the exocrine tissue portion are trapped in an electromagnetic field, thus providing effluent islets of a high degree of purity. In this study our aim was to adapt this principle to human islet preparations. In this context our prime interest was focused on a lectin suitable for human pancreatic tissue. Of 19 different lectins tested, only 1, Wisteria floribunda agglutinin (WFA), is suitable, as shown by immunofluorescence, MMS-lectin binding, and magnetic separation.

Cell Separation↗
PubMed1989

Immunoelectron microscopic localization of MHC structures in isolated pancreatic rat islets.

An immunogold-silver enhancement technique, which combines effective labeling of viable isolated islets with the ultrastructural resolution of cytological details, was applied in electron microscopy to identify major histocompatibility complex (MHC) structures on islet cells. Incubation of freshly isolated islets from CAP (RT1c) and LEW (RT1l) rats with OX18, an MHC class I antibody, showed strong positive reactivity in macrophages and/or dendritic-like cells (M0-DCs) and vascular endothelial cells (VEs) and a comparatively weaker reactivity in endocrine alpha-, beta-, and delta-cells. With MHC class II antibody OX6 (anti-I-A), M0-DCs were strongly labeled in both rat strains on the surface and on internal structures. Three of five particularly high titered batches of OX6 revealed MHC class II expression on VE and beta-cells. Four days of in vitro culture in combination with a high concentration of glucose and interferon-gamma induced strong enhancement of MHC class I structures and, to a lesser extent, class II structures on beta-cells.

Animals↗
PubMed1989

[Changed chemosensory identity following experimental bone marrow transplantation: recognition by another species].

A new paradigm was constructed using rats as trained animals in order to examine changes appearing in the scent of mice after a fully allogeneic bone marrow transplantation (BMT). Two rats (CAP), deprived of water 20 hours before each training session, were trained in an olfactometer to discriminate two fully allogeneic mice strains (C3H, C57) via their urine odors. Reinforcement for identification of the St was provided by a drop of water. After discrimination was built-up, the reinforcement schedule was reduced stepwise in defined blocks of trials, in order to allow transfer-of-training tests, to which urine samples of allogeneic reconstituted BMT-chimeras (C57----C3H and C3H----C57) were submitted. It was confirmed that the strain-specific urine odors of the recipients were changed by a fully allogeneic BMT. The results also showed that the urine odors of allogeneic reconstituted chimeras differed from the specific urine odor of the donor strain.

Animals↗
PubMed1989

[Exchange of the hematopoietic system changes chemosensory identity].

It has been shown that the scent of mice changes after a bone marrow transplantation. Previously obtained results were re-examined with a new design that rules out possible avoidance learning effects. Two BALB/c mice, deprived of water for 20 hours each day, were trained in a Y-maze to discriminate two fully allogeneic mice strains (C3H = S+ and C57 = S-) via their urine odor. Reinforcement for correct choice was provided by a drop of water. Urine samples of different fully allogeneic mice strains (C3H, C57 and A/J) and bone marrow transplanted chimeras (C57----C3H and C3H----C57) were submitted to different "transfer of training" -tests. In the conditioning paradigm used, the animals learn to identify S+ and also do not avoid S- after the training. The urine odor of the C57----C3H-chimeras was found to be different from the strain-specific urine odor of C3H animals. Since this change could not be found in syngeneic transplanted chimeras, it is concluded that it is caused by the graft. The experiments failed, however to demonstrate that the urine odor of allogeneic chimeras is constituted solely by donor- and recipient-specific components.

Animals↗
PubMed1988

Morphologic characteristics of developing osteoarthrotic lesions in the knee cartilage of STR/IN mice.

The development of osteoarthrotic cartilage lesions in the knee joints of male STR/IN mice was studied with respect to their histologic appearance and their various localizations in the joint. Spontaneous articular cartilage degeneration on the medial portion of the tibial plateau was considered to be the initial event. Continued loss of cartilage subsequently led to a pronounced instability of the knee joint, with a varus deformity. This was followed by medial patellar luxation with corresponding osteoarthrotic lesions at the facies patellaris femoris. The most marked osteoarthrotic cartilage degeneration developed on the medial tibial condyle and at the facies patellaris femoris of the femoropatellar joint. Histologic examination of the osteoarthrotic defects in these two regions revealed distinct morphologic differences with respect to formation of chondrocyte clusters, tendency to regeneration, and proliferation reactions. Lectin binding experiments in normal articular cartilage revealed regional differences regarding the presence or absence of keratan sulfate in the extracellular matrix. The lack of keratan sulfate in tibial cartilage might reflect its tendency to degenerate spontaneously. It is therefore suggested that male STR/IN mice are particularly useful for studying two different types of osteoarthrosis, one due to a biomechanically induced instability (patellar luxation) and one due to biochemical changes (absence of keratan sulfate) of still unknown pathogenesis.

Animals↗
PubMed1988

Lectin-binding in normal and osteoarthrotic articular cartilage from STR/1N-mouse knee joints.

Fluorescein-isothiocyanate (FITC) labeled lectins were used to study the distribution pattern of specific binding-sites in histological sections of normal and osteoarthrotic articular cartilage from the mouse knee joint. Male inbred mice of the STR/1N-strain develop spontaneous arthrotic articular cartilage lesions on the medial condyle of tibia and femur. The varus-deformity of the knee joint leads to a recurrent medial patellar luxation with osteoarthrotic defects on the medial part of the facies patellaris femoris. It was demonstrated that the lectin staining pattern of osteoarthrotic articular cartilage, especially on the facies patellaris femoris, was different from that of normal articular cartilage. The differences in lectin staining corresponded to those observed between normal and fibrillated articular cartilage from human patellae. The normal articular cartilage of the mouse knee joint possessed lectin binding-sites for Concanavalin A (ConA) and wheat germ agglutinin (WGA), but not for Ulex europaeus agglutinin (UEA), soy bean agglutinin (SBA) and peanut agglutinin (PNA). In addition to the completely changed distribution pattern of ConA and WGA in osteoarthrotic cartilage, SBA, PNA and UEA developed distinct staining patterns particular to the fibrillated areas of arthrotic cartilage. The increased lectin-binding to arthrotic articular cartilage may be due to unmasking of sugars in the course of bondage breakdown in fibrillated cartilage or the production of pathological glycoproteins. It is evident that lectins can demonstrate minute differences between normal and arthrotic cartilage and it is concluded, therefore, that lectins are sensitive and specific tools for the study of degenerative joint diseases.

Animals↗
PubMed1988

Carbohydrate-specific human heterophile antibodies in normal human sera that react with xenogeneic cells.

Human heterophile antibodies (HHA) that are present in normal human sera (NHS) play an important role in hyperacute xenograft rejection. The aim of this study was to analyze the occurrence, mode of action and molecular specificity of HHA in NHS that are directed against xenogeneic lymphocytes (isolated from mouse, rat, guinea pig, rabbit, cattle and pig) and isolated rat pancreatic islets. All sera contained variable amounts of HHA that killed the target cells via the classical complement pathway. The cytotoxic activity of these HHA was specifically inhibited by certain carbohydrates (alpha-D-melibiose, beta-lactose, beta-gentiobiose, beta-cellobiose, D-mannose, N-acetyl-beta-D-mannosamine and alpha-D-rhamnose) and by rat IgM. By means of affinity chromatography with immobilized inhibitors we obtained an antibody preparation of mainly IgG type from NHS (up to 3.5 mg/10 ml serum) that reacted strongly with rat lymphocytes and isolated rat pancreatic islets. Though thus far residual xenospecific antibody activity has remained in the sera even after multiple affinity chromatography, these data suggest that specific elimination of HHA is feasible and that it may be thus possible to overcome a major obstacle to xenotransplantation.

Antibodies, Heterophile↗
PubMed1988

Human monoclonal antibody against human lymphocytic cells. A human monoclonal antibody that reacts preferentially with human lymphocytic cells.

Human monoclonal antibodies may replace human or xenogeneic antisera and mouse monoclonal antibodies for therapeutic applications. The human IgM monoclonal antibody Ha6D3 was produced after in vitro immunization and fusion with a mouse myeloma. Its reactivity against human normal and leukemic cells was investigated in cytotoxicity assays, and the antigen distribution in normal tissues was investigated with biotinylated Ha6D3 and avidin-peroxidase complexes. It was shown that Ha6D3 reacts preferentially with human lymphocytic cells. Only moderate reactions with epithelial cells in some organs were observed in cryostat sections, but because of poor accessibility in vivo these reactions were considered to be negligible.

Antibodies, Monoclonal↗
PubMed1988

Measurement of rat insulin. Enzyme-linked immunosorbent assay with increased sensitivity, high accuracy, and greater practicability than established radioimmunoassay.

Total immunoreactive insulin (IRI) is conventionally determined by radioimmunoassays. IRI measurement in rats can be made more sensitive, accurate, and practical, as demonstrated by a new modified enzyme-linked immunosorbent assay (ELISA). It is characterized by indirect binding of an anti-insulin antibody by an antiglobulin antibody and uses the principle of competitive saturation. In this ELISA, IRI can be determined in a wide range of concentrations, corresponding to the standards. The standard curve ranges from 100 to 0.049 ng/ml IRI (1 ng/ml approximately 23.4 microU/ml approximately 172 pM rat insulin). The statistical analysis shows between- and within-assay coefficients of variation of less than or equal to 15%.

Animals↗