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Biomedical subjects

E Hinzová

Publications and source records attributed to E Hinzová.

11 recordsLinked to original sources

Inhibition of adhesion of F9 embryonal carcinoma cells to substratum by a novel monoclonal antibody, TEC-05, reactive with a developmentally regulated carbohydrate epitope.

Embryonal carcinoma cells carry on their surfaces carbohydrate antigens that are also expressed in early embryonic cells. We report here the expression and properties of a new developmentally regulated carbohydrate epitope, which is defined by a monoclonal antibody TEC-05. This antibody was generated by immunization of a rat with mouse embryonal carcinoma cells P19S1801A1. By immunofluorescence, the TEC-5 epitope was first detected on 8-cell-stage mouse embryos and was present on all subsequent stages of preimplantation development. Absorption analysis revealed that TEC-5 epitope was expressed only on a limited number of adult mouse tissues. In the direct radioantibody binding assay, TEC-05 reacted strongly with OTF9-63 cells and with some of the mouse embryonal carcinoma cell lines tested. Its reaction with differentiated cell lines was weak or undetectable. In the course of differentiation of OTF9-63 cells induced by retinoic acid, the epitope disappeared with the onset of morphological differentiation. The binding of the antibody to OTF9-63 cells was inhibited to 50% by 10-50 microM N-acetyllactosamine and lactose. Immunolabelling of extracts from OTF9-63 cells separated by sodium-dodecyl-sulfate (SDS) polyacrylamide gel electrophoresis revealed that TEC-5 epitope was carried by high-molecular-weight glycoconjugates (molecular weight greater than 100,000). Molecules, isolated from [3H]-fucose-labelled OTF9-63 cells by indirect immunoprecipitation with TEC-05 antibody, were degraded by extensive pronase digestion or mild alkaline treatment to large carbohydrate chains that were excluded from a Sephadex G-50 column. Direct evidence that TEC-05 antibody bound to embryoglycan was obtained using a modified Farr's assay. The antibody was found to inhibit adhesion of F9 and OTF9-63 cells to substratum. The inhibitory effect, which could be abrogated by lactose, seemed to be specific, because another IgM monoclonal antibody which also binds to embryoglycan had no effect. Combined data indicated that TEC-05 antibody recognizes a carbohydrate epitope which is involved in cell-substratum adhesion of F9 cells and which provides a new marker for structure-function studies of stage-specific embryonic antigens.

Animals↗

Quantitative estimation of HLA-A and HLA-B antigens carrying the Bw4 supertypic specificity in human peripheral blood lymphocytes.

A monoclonal antibody specific for HLA-Bw4 was employed for the quantitative estimation of class I antigens on human peripheral blood lymphocytes and on the human macrophage cell line U 937. The epitopes reactive with the HLA-Bw4-specific antibody, which are present on different antigens coded by the HLA-A or the HLA-B locus, were characterized in terms of equilibrium and kinetic binding parameters. The level of expression of class I antigens on human lymphocytes was found to be in direct proportion to the gene dose. Variations between donors of the same phenotype were of minor importance. Estimation of the association constant, association rate constant, and half-life of dissociation for the interaction of the antibody with lymphocytes heterozygous for either HLA-A24, HLA-A32, HLA-A9, or HLA-Bw4 strongly suggested that the public antigenic determinant shared by these antigens is identical.

Antibodies, Monoclonal↗

Supertypic HLA-Bw4 antigen detected by a new monoclonal antibody.

A monoclonal antibody, HLAO1, was prepared by immunizing BALB/c mice with human lymphocytes of known HLA genotype. The monoclonal antibody is cytotoxic, of the IgG 2b isotype and binds to a protein of Mr 43,000 noncovalently associated with the beta 2-microglobulin. Genetic analysis proved complete concordance in the expression of the antigenic determinant defined by this monoclonal antibody and the presence of a supertypic (public) antigen, the HLA-Bw4. As expected, the monoclonal antibody also reacted with the antigens of HLA-A locus, A23, A24, and A32. The computer analysis of HLAO1 antibody binding to the Bw4/Bw6 heterozygous lymphocytes gave the approximate number of antigenic determinants, n = 2.2 X 10(4)/cell, and Ka = (5.9 +/- 0.8) X 10(9) M-1. Besides its potential immunochemical applications, the HLAO1 monoclonal antibody can become a useful tool in routine cytotoxicity typing of HLA antigens on peripheral lymphocytes.

Animals↗

HLA-D antigens in a Czech population and their association with HLA-DR antigens.

HLA-D specificities Dwl through 11 and HLA-DR antigens DRwl through WIA8 were studied on a panel of 101 unrelated individuals of the Prague HLA panel. THe HLA-D specificities were defined by means of the HLA-D homozygous typing cells in MLC test, the HLA-DR specificities were tested on B cells using VIIth Histocompatibility Workshop antisera. In the panel tested, the antigen and gene frequencies of eleven HLA-D determinants, the associations between the HLA-B and HLA-D antigens as well as between the HLA-D and HLA-DR antigens were determined. Significant associations between some of the HLA-D and HLA-DR antigens were found.

Czechoslovakia↗

HLA--Dw3 in Sjögren's syndrome.

The increased frequency of HLA-B8 in Sjögren's syndrome was recently reported independently by Gershwin et al. (1975) and Iványi et al. (1976). The association of HLA-B8 antigen with other diseases, characterized generally by impaired immunologic reactivity, has been shown (see Svejgaard et al. 1975, Dausset & Hors 1975), and in some of these diseases MLC typing has revealed the strong association also with the Dw3 determinant. The degree of association with each of the two, B8 and Dw3, varied between different diseases, in some of them having been reported to be higher for antigen B8 (Möller et al. 1976) and in the others for the determinant Dw3 (Thomsen et al. 1975, 1976). In some of the diseases, the association was found to be equally strong for both determinants (Solheim et al. 1976, Thorsby et al. 1975). In this communication, we describe the typing of HLA-Dw3 in 29 patients with Sjögren's syndrome (Ss). They represent part of the experimental group reported in a previous study (Iványi et al. 1976).

HLA Antigens↗

The distribution of HLA-d (MLC) locus determinants in the Czech population.

The distribution of 8 HLA-D determinants in the group of 83 unrelated individuals of the Czech population was ascertained by means of homozygous typing cells using the mixed lymphocyte culture test. In the panel tested, the gene frequencies of HLA-D determinants, the distribution of HLA-D phenotypes and the magnitude of the linkage disequilibrium between the determinants of the HLA-B and HLA-D loci were determined.

Czechoslovakia↗

Mouse lymph node activating factor released during contact of HL-A-different blood lymphocytes.

Ten-fold concentrated supernatants from the 4-hr mixed cultures of lymphocytes from two humans different at two haplotypes were inoculated s.c. into the hind footpad of mice. Twenty-four to forty-eight hours later, the cellularity and the synthesis of ribonucleic acid increased significantly in the popliteal lymph nodes draining the site of injection. The supernatants from cultures of HL-A-identical cells of cells different at one haplotype lacked this activity. The results suggest that human imcompatible lymphoid cells upon mutual contact liberate a soluble mediator capable of activating the draining lymph nodes; i.e., it is able to trap cells from the circulation and to stimulate them. The mediator is species-nonspecific; the activity produced by human lymphocytes can be tested on mice.

Animals↗

The role of the histocompatibility-2-linked Ss-Slp region in the control of mouse complement.

The Ss-Slp region of the mouse genome is located between the two loci, H-2K and H-2D, of the major histocompatibility system. It controls the level of a serologically detectable serum globulin, the Ss protein, and its allotypic forms, Slp(a) and Slp(o). Evidence is presented that the Ss-Slp region is involved in control of the complement system. Mice of congenic resistant strains differing only at the H-2 gene complex show differences in complement levels. A comparison of H-2 recombinants demonstrated that complement level correlates with the Ss-Slp genotype and not with the H-2K, H-2D, or Ir genotypes. Among the 13 congenic strains tested, those with the Ss(h)Slp(a) allele had higher complement levels than those with the Ss(h)Slp(o) or Ss(l)Slp(o) alleles. Mice with recombinant H-2 haplotypes had complement levels similar to those of the parental strain that provided the Ss-Slp segment. It was also shown that anti-Ss serum inhibits complement activity of mouse serum in vitro.

Animals↗