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Biomedical subjects

H Binz

Publications and source records attributed to H Binz.

At least 55 records · Page 3Linked to original sources

Epitope mapping by chemical modification of free and antibody-bound protein antigen.

A monoclonal antibody bound to a protein antigen slows the rate of chemical modification of amino acid residues located at the epitope. By comparing the degree of acetylation of 18 lysine and 7 threonine residues in free and antibody-bound horse cytochrome c, a discontiguous, conformational epitope was characterized on this protein antigen. The new approach is particularly suitable to probe discontiguous and conformational epitopes, which are difficult to analyze by other procedures.

Amino Acid Sequence↗

Studies of monoclonal antibodies specific for major histocompatibility complex products of the rat. IV. Production and characterization of polyclonal and monoclonal anti-anti-idiotypic antibodies.

Polyclonal syngeneic, allogeneic, and xenogeneic and monoclonal syngeneic anti-anti-idiotypic antibodies have been produced against previously described monoclonal anti-idiotypic antibodies with specificity for monoclonal RT1 alloantigen-specific antibodies. The anti-anti-idiotypes could again be shown to be highly specific for the monoclonal anti-idiotype used for the induction of the anti-anti-idiotypic antibodies and to carry the same, or a very similar, idiotype as the original monoclonal idiotypic antibody used to induce the monoclonal anti-idiotypic. Among the 30 syngeneic and allogeneic and the five xenogeneic polyclonal anti-anti-idiotypic antisera and the three monoclonal anti-anti-idiotypes, only one polyclonal antiserum showed binding capacity to the corresponding RT1-encoded antigenic determinants on spleen cells. All the other antibodies were idiotypic but not antigen binding.

Animals↗

Monoclonal antibodies specific for Sendai virus. I. Production and characterization of monoclonal antibodies.

Twelve monoclonal antibodies specific for Sendai virus were prepared by fusing immune LOU rat spleen cells with Y3 or FO myeloma cells. Six antibodies bound the viral glycoprotein HN, and six the viral protein F. Among the six HN-specific monoclonal antibodies, five reacted with the very same epitope and inhibited viral haemagglutination. Two antibodies against the F protein recognized the same epitope, but all the others reacted with different epitopes. All monoclonals were characterized with regard to specificity, biological function, epitope recognition, isotypes, and pI.

Antibodies, Monoclonal↗

Monoclonal antibodies specific for Sendai virus. II. Production of monoclonal anti-idiotypic antibodies.

The monoclonal antibodies A and B specific for the HN molecule of Sendai virus were used to induce polyclonal and monoclonal anti-idiotypic antibodies. No response was observed in allogeneic Lewis rats, low responses in syngeneic LOU rats, and high responses in allogeneic BN rats and xenogeneic Balb/c mice. The monoclonals A and B share a similar or identical idiotype, since polyclonal anti-idiotypic antisera to antibody A cross-reacted completely with antibody B and vice versa. The same was found with the three monoclonal anti-idiotypes 1, 2, and 3 elicited in a BN rat or in Balb/c mice. None of the polyclonal or monoclonal anti-idiotypes reacted with other monoclonal antibodies specific for Sendai virus, even when these recognized the same epitope as antibodies A and B. The monoclonal anti-idiotypic antibodies could be used to induce anti-Sendai antibodies in BN rats.

Antibodies, Anti-Idiotypic↗

Cellular location of 'liver-specific' antigen F.

Three mouse monoclonal antibodies against Type 1 murine 'liver-specific' antigen F and one rat monoclonal antibody against human antigen F were produced. These monoclonals recognize antigenic determinants common to Type 1 and Type 2 mouse and human antigen F. By immunolabelling with beta-galactosidase, antigen F could be demonstrated in the cytoplasm of hepatocytes, tubules of the kidney, and perikarya of neurons of the central nervous system. F antigens isolated from liver, kidney, and brain all have molecular weights of around 40,000.

Animals↗

Use of donor-specific T-cell lines for monitoring of human allograft recipients. I. Demonstration of IgG binding to autologous TCL.

Donor-specific and highly cytotoxic T-cell lines (TCL) as well as lectin-induced TCL were established from pretransplant lymphocytes of 6 cadaveric renal allograft recipients. These TCL were used in the 125I-staphylococcus protein A assay to detect IgG antibodies in pre- and posttransplant sera of these patients preferentially binding to autologous donor-specific TCL. Such antibodies were detected in pretransplant sera from 4 of these 6 allograft recipients. Antibody levels in these 4 patients and in 1 additional case who became positive after transplantation further increased during acute cellular rejection episodes. They disappeared after successful treatment but remained elevated until transplantectomy for treatment of irreversible rejection in 1 case. IgG antibodies binding to autologous lectin-induced TCL were detected in only 1 patient and exhibited a pattern clearly different from those binding to donor-reactive TCL. Although attempts to define the antigenic specificity of the autoantibodies binding to donor-specific TCL by genetical and biochemical means has remained unsuccessful so far, the demonstration of their relationship to in vivo expansion of donor-reactive immune cells deserves further attention.

Adult↗

Studies of monoclonal antibodies specific for major histocompatibility complex products of the rat. I. Production and characterization of monoclonal antibodies.

Sixteen monoclonal antibodies against antigens coded for by the RT1 complex of the rat have been produced. Fourteen are specific for the a haplotype: six recognize class II and eight class I antigens. Two are specific for the 1 haplotype, one reacting with class I and the other with class II antigens. By means of these monoclonal antibodies four independent clusters of antigens for class I antigens of the a haplotype and three for class II antigens could be defined. The three antigenic sites of class II antigens reside on the same heterodimer. The monoclonals described here are characterized with regard to Ig class and subclass, pI, and complement-activating capacity.

Animals↗

Studies of monoclonal antibodies specific for major histocompatibility complex products of the rat. II. Production and characterization of xenogeneic, allogeneic, and syngeneic polyclonal anti-idiotypic antibodies.

Polyclonal anti-idiotypic antibodies against monoclonal antibodies with specificity for RT1-encoded antigens were induced in syngeneic, allogeneic, and xenogeneic hosts. The immune response to the idiotypic determinants on monoclonal antibodies was T-cell-dependent. The anti-idiotypic antibodies, independent of whether they were induced in syngeneic, allogeneic, or xenogeneic (after proper absorption) hosts, showed an exquisite specificity for the monoclonal antibody used for the induction. No cross-reactivity with other monoclonal antibodies could be observed.

Animals↗

Studies of monoclonal antibodies specific for major histocompatibility complex products of the rat. III. Production and characterization of syngeneic and allogeneic monoclonal anti-idiotypic antibodies to monoclonal alloantibodies.

Sixteen monoclonal anti-idiotypic antibodies were prepared against RT1 antigen-specific monoclonal antibodies by means of an inhibition assay. Three (110-3, 112-89, and 133-4) were produced against antibody 3-8C2 (19), three (178-60, 181-14, and 184-38) against antibody 11/23/5, and three (144-3, 144-33, and 146-31) against antibody 1-5A4. One monoclonal anti-idiotypic antibody (21-39) was directed against antibody 1-8D2 and six (CE1-3, CE2-1, CE7-2, CE8-46, CE9-35, and CE10-2) against antibody 3-12A1. The anti-idiotypic antibodies were all highly specific for the respective monoclonal antibody used for the induction of the anti-idiotype and did not cross-react with other monoclonal antibodies.

Animals↗

Studies of the invasiveness of the chemically induced mouse sarcoma FS9. I. Monoclonal antibodies to a 37,000 dalton membrane glycoprotein inhibit invasion of fibroblasts in vitro.

Monoclonal antibodies against the mouse sarcoma FS9 were prepared. Antibodies recognizing a 37,000 dalton glycoprotein on FS9 tumor cells inhibit invasion by FS9 sarcoma cells of chicken heart fibroblasts in vitro as assessed by Abercrombie's confronted explant assay. Antibodies to other membrane proteins of FS9 tumor cells failed to inhibit invasiveness of FS9 sarcoma cells. The 37,000 dalton glycoprotein, which is neither a histocompatibility antigen nor a gp37 glycoprotein of Rous sarcoma virus nor the MEP described by Gottesman, is present on the surface and in the cytoplasm of FS9 sarcoma cells. The plasma membrane of the non-invasive mouse cell line L929 contains only low concentrations of the 37,000 dalton antigen. Hence, a relationship apparently exists between the increased concentration of this protein on the surface of FS9 cells and their invasiveness.

Animals↗

Invasive behavior of mouse sarcoma cells is inhibited by blocking a 37,000-dalton plasma membrane glycoprotein with Fab fragments.

Abercrombie 's confronted explant technique was used to study the role of tumor surface antigens in malignant invasion. Plasma membranes were isolated from mouse sarcoma cells ( FS9 ) and a mouse cell line (L929) of the same H-2 haplotype. FS9 cells are highly invasive when confronted with chicken heart fibroblasts, whereas the L929 cells are not [ Abercrombie , M. (1979) Nature (London) 281, 259-262]. The FS9 plasma membranes contained significantly higher concentrations of a 37,000-dalton glycoprotein. When antiserum directed against FS9 plasma membranes was preabsorbed with L929 cells, the antibodies remaining reacted predominantly with the 37,000-dalton antigen. Fab fragment prepared from the preabsorbed antiserum inhibited the invasion of chicken heart fibroblasts by FS9 cells. Fab prepared from a monoclonal antibody directed against the 37,000-dalton antigen also inhibited invasivity , whereas monoclonal antibodies reacting with two other FS9 cell surface antigens did not. The results imply a relationship between the increased concentration of the 37,000-dalton glycoprotein on the surface of the FS9 cells and their invasivity .

Animals↗

Studies on chemically induced rat tumors. II. Partial protection against syngeneic lethal tumors by cloned syngeneic cytotoxic T lymphocytes.

In the present article we describe studies on a chemically induced sarcoma in DA rats. This tumor expresses a unique antigen which can be demonstrated by both syngeneic antibodies and cytotoxic T cells. We have established cytotoxic T-cell lines (CTLs) specific for the tumor and with high efficient killing capacity in vitro. When testing for the ability of such CTLs to inhibit tumor out-growth in vivo, we found that they had to be inoculated together with the tumor and in the presence of T-cell growth factor to provide any significant degree of protection. We thus believe not only that there is a requirement for addition of CTL-stimulating lymphokines in vivo, but also that the CTLs fail to move from one site in vivo to attack relevant tumor cells at another site. No evidence was obtained that the CTLs gradually could acquire such migratory ability in vivo.

9,10-Dimethyl-1,2-benzanthracene↗

Two independent receptors allow selective target lysis by T cell clones.

Dimethylbenzanthracene-induced P1 sarcoma cells induce P1-specific antibodies in syngeneic DA rats. Antiidiotypic antibodies of specificity DA anti-(DA anti-P1) were induced against the tumor-specific antibodies and used to restimulate P1-primed DA T cells in vitro. Using antiidiotypic antibodies and T cell growth factor, P1-specific cytotoxic DA T cell clones were established by limiting dilution and kept in vitro. Two of these clones acquired during culture periods in addition to the P1 specificity lytic activity towards natural killer (NK) targets YAC-1 or K562. Cold target inhibition experiments showed that the very same cytotoxic T cells kill P1 and NK targets. Antiidiotypic antibodies of specificity DA anti-(DA anti-P1) inhibited cytotoxicity against P1 but not against YAC-1 or K562. We conclude that two independent receptors are located on these double-reactive T cell clones, one that is idiotypic and antigen-specific, and another displaying the binding profile of NK cells.

Animals↗

Studies on chemically induced tumors in rats: I. Heterogeneity of tumor cells and establishment of syngeneic, tumor-specific cytotoxic T cell clones.

Sarcoma P1 was induced in DA rats by DMBA. Anti-P1 antibodies were produced in DA rats, purified via fixed tumor cells and used to induce anti-idiotypic antibodies in syngeneic rats. The anti-idiotypic antibodies were used to generate cytotoxic, P1 specific DA T cells in vitro. These cytotoxic T cells and P1 tumor cells were cloned by limiting dilution. Using the DA anti-P1 specific cytotoxic T cell clones, we were able to characterize 2 types of P1 tumor cell clones, namely those which were susceptible and those which were resistant to the P1 specific cytotoxic T cells. Cytotoxic T cell injected i.v. into syngeneic DA rats could not prevent the development of lethal P1 tumors.

Animals↗

Monoclonal antibodies directed to human insulin-like growth factor I (IGF I). Use for radioimmunoassay and immunopurification of IGF.

Mouse hybridomas secreting antibodies to human insulin-like growth factor I (IGF I) were produced by fusion of spleen cells of hyperimmunised mice with FO mouse-myeloma cells. Eight clones producing antibodies against human IGF I have been isolated, two of which have been characterised. One was used in a radioimmunoassay, the other for immunopurification of IGF.

Acromegaly↗

Induction or elimination of tumor-specific immunity against a chemically-induced rat tumor using auto-anti-idiotypic immunity.

DA rat sarcoma P1 and P2 were induced by dimethylbenz(a)anthracene. A tumor-specific immune response of DA rats against P1-tumor cells could be demonstrated at the humoral and cellular level. DA anti-P1 antibodies were purified on fixed P1-tumor cells and used as auto-immunogen in DA rats for the production of anti-idiotypic antibodies. Such anti-idiotypic antibodies could be demonstrated by using a solid-phase radioimmunoassay and by their ability to induce secondary type of DA anti-P1 response in vitro. In addition, such antibodies were able to induce cytotoxic T lymphocytes capable of eliminating P1-tumor cells but not control tumor cells. In some of the auto-immunized DA rats enhanced P1-tumor growth could be observed, indicating that the anti-idiotypic immune response had led to a selective ablation of idiotypic, potential anti-P1 reactive T cells.

9,10-Dimethyl-1,2-benzanthracene↗

An immunoglobulin V gene polymorphism in the rat.

Anti-phenyloxazolone (phOx) antibodies of different AVN rats (primary response) share several isoelectric focusing bands. These bands were not shared by antibodies of some other rat strains, including DA. An anti-idiotype reagent was prepared (in rabbits) that bound radioactive anti-phOx antibodies of AVN rats but not normal AVN immunoglobulin. This binding was strongly inhibited by AVN anti-phOx antisera, but not by AVN anti-BOC-p-azobenzene arsonate-tyrosine antisera or DA anti-phOx antisera. Anti-phOx antisera of (AVN X DA)F1 rats were also strongly inhibitory indicating the presence of the idiotype (Ox-r1). Antisera of backcross rats (AVN X DA) X DA either resembled F1 hybrid sera (31 rats) or DA sera (23 rats). The data suggest that the presence of idiotype Ox-r1 is controlled by one gene, or genes linked to each other. The gene(s) is not linked to the Ig kappa chain locus. It may be a V gene of the Ig H chain.

Animals↗