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Biomedical subjects

W Müller-Ruchholtz

Publications and source records attributed to W Müller-Ruchholtz.

At least 91 records · Page 5Linked to original sources

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

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

Production and applications of new monoclonal antibodies against human lymphocyte antigen-A and -B antigens.

Monoclonal antibodies (MAbs) against HLA antigens can give better information about the serology and biochemistry of the human major histocompatibility complex (MHC) than HLA antisera obtained from pregnant women. To increase the very limited panel of MAbs against HLA we immunized mice with human cultured cell lines and fused their spleen cells with the Ag8-653 myeloma. We produced MAbs against HLA-A1, A2/w69, A2/28, A2/11/25/26/28/29/30/31/33/34, A25/32, B7/22, and B13. The best dilution medium to store the MAbs in Terasaki plates was RPMI 1640 supplemented with 7% bovine serum albumin. The MAbs are also excellent reagents to investigate public HLA antigens and to stain HLA antigens in cryosections of transplanted organs.

Animals

Covalent conjugates of monoclonal antibody and cobra venom factor mediate specific cytotoxicity via alternative pathway of human complement activation.

Because of lack of unspecific toxicity autologous serum is the optimal source of complement for in vitro cell depletion in human bone marrow transplantation. Covalent binding of cobra venom factor to an antibody leads to a specific complement activating conjugate with cytolytic potential independent of the capacity of the antibody to fix complement itself. Therefore this conjugation allows the use of various sera, including human as the source of complement. The kinetics of cobra venom factor-initiated cytolysis is compatible with in vitro treatment of bone marrow prior to transplantation.

Antibodies, Monoclonal

Comparison of gamma-interferon and alloantigen-induced T cell factors in the induction of MHC class II antigen expression and in the modulation of the immunogenicity of lymphocyte-free rat bone marrow.

In a rat model, we compared the effects of various soluble products released by T cells, such as the unspecifically acting gamma-interferon or the newly detected alloantigen-induced factors that specifically act on nonlymphoid, hemopoietic bone marrow cells. We found two types of reactivity patterns with regard to the induction of MHC class II antigen expression on these cells. The very same patterns could be demonstrated when we investigated the modulation of their stimulatory capacity, i.e., their immunogenicity in a T proliferation assay. These findings are discussed in relation to the increasing incidence of immunologically mediated graft rejections in clinical bone marrow transplantation following T cell purging.

Animals

Transplantation of aortic and pulmonary allografts, enhanced viability of endothelial cells by cryopreservation, importance of histocompatibility.

Fresh heart valve allografts were preserved at 4 degrees C for 14 days, cryopreserved and stored for 63 days, and studied for endothelial viability and antigenicity, in order to obtain some information on the immunobiological status of allografts before transplantation. The surgical technique described by Ross for subcoronary position is preferred and briefly outlined. Four explanted incompatible allografts were studied by light and scanning electron microscopy and immunohistochemistry to assess the immunological reactions and tissue changes that occurred between 9 days and 16 weeks postoperatively. Valve leaflet motion and opening velocities were studied by echocardiography postoperatively to establish a baseline with which to distinguish early leaflet degeneration. Distensibility of the aortic annulus was studied postoperatively by supraaortic angiography to justify one of the goals of reconstruction of the aortic root with allografts. Antibiotic preserved allografts at 4 degrees C showed no viable endothelial cells after 8 days while the cryopreserved allografts demonstrated a high rate of viable endothelial cells capable of expressing surface antigens (HLA class I and II). Although the valve explants showed focal mononuclear cell infiltrations with T-lymphocytes, the allografts healed in place. The "classic" findings of rejection could not therefore be demonstrated. In summary, cryopreserved valve allografts, like the fresh, are antigenic. It is therefore recommended to use compatible valve grafts, when possible, which might be a positive step to improve the functional longevity of valve allografts. Immune response after valve allograft transplantation does not cause acute valvular dysfunction but rather chronic tissue changes which might lead to early degeneration of the allograft. The opening velocities of preserved aortic and pulmonary allografts were normal at 3-4 years postoperatively irrespective of histocompatibility. Echocardiography might be a useful tool to detect early degenerative changes of incompatible valve leaflets. The aortic root is distensible after allograft transplantation.

Adolescent

Role of lymphocytes in the modulation of the immunogenicity of bone marrow: a rat model.

In a rat model we describe the role T lymphocytes play in the immunogenicity of bone marrow. Mainly they modulate the MLC-like stimulatory cell potential and the MHC class II antigen expression of nonlymphoid bone marrow cells via the release of newly detected specific soluble factors and immunologically unspecific ones. These findings are discussed in relation to the graft rejection encountered in clinical bone marrow transplantation following certain T depletion protocols.

Animals