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Biomedical subjects

H Binz

Publications and source records attributed to H Binz.

At least 109 records · Page 6Linked to original sources
PubMed1975

Shared idiotypic determinants on B and T lymphocytes reactive against the same antigenic determinants. III. Physical fractionation of specific immunocompetent T lymphocytes by affinity chromatography using anti-idiotypic antibodies.

Anti-idiotypic antibodies made against antigen-binding receptors on T lymphocytes with specificity for certain Ag-B locus antigens selectively react with T lymphocytes with potential immune reactivity against the very same Ag-B antigens. This was shown by affinity chromatography of normal Lewis T lymphocytes on anti-Ig columns after contact with the relevant anti-idiotypic antiserum. Here, it could be shown that incubation of the cells with an anti-(Lewis-anti-BN) antiserum caused subsequent selective retention of potential graft-vs.-host (GvH)-reactive cells against BN on the anti-Ig column, whereas Lewis T cells with reactivity against DA or August (Au) (carrying distinct Ag-B antigens in comparison to BN) passed through. The retained cells could be eluted and shown to display highly increased reactivity against BN with virtually no reactivity left against DA or Au antigens. Analogous results were obtained using an anti-(Lewis-anti-DA) antiserum. The anti-idiotypic antibodies can be used in fluorescent antibody tests to directly visualize the idiotype-positive cells. Using the separation design described above we analyzed selectively enriched or deleted T lymphocytes for presence of idiotypic cells as well as specific GvH reactivity. A highly significant positive correlation was found between percentage of a given idiotype in a population of T cells and the relevant GvH potential of the same T cells that can be visualized are indeed the very same T cells that express immune reactivity against the expected antigens. The present data would thus directly demonstrate the existence of a largely nonoverlapping population of immunocompetent T cells capable of reacting against the various Ag-B locus antigens in the rat. Highly purified, functionally intact immunocompetent T lymphocytes with restricted immune reactivity can thus be produced from normal lymphocyte populations for further analysis.

Animals↗
PubMed1975

Shared idiotypic determinants on B and T lymphocytes reactive against the same antigenic determinants. I. Demonstration of similar or identical idiotypes on IgG molecules and T-cell receptors with specificity for the same alloantigens.

Antigen-binding receptors on T lymphocytes and IgG antibodies with the same antigen-binding specificity as the T-cell receptors display shared or identical idiotypes. This was shown using a system where adult F1 hybrid rats between two inbred strains were inoculated with T lymphocytes from one parental strain. Such F1 hybrid rats produce antibodies directed against idiotypic determinants present on IgG alloantibodies, produced in the T donor genotype strain and with specificity for the alloantigens of the other parental strain. The idiotypic nature of the F1 antialloantibody serum against the parental alloantibodies was demonstrated both by indirect hemagglutination tests or by gel diffusion using alloantisera with different specificity as targets. Furthermore, the F1 anti-T-lymphocyte sera could be shown to contain antibodies against idiotypic parental T lymphocytes as well. This was shown by the capacity of the antisera, in the presence of complement, to wipe out the relevant parental T-cell reactivity against the other parental strain (as measured in MLC or GVH) whilst leaving the T-lymphocyte reactivity against a third, unrelated allogeneic strain intact. These findings demonstrate that F1 hybrid rats inoculated with parental T lymphocytes make anti-idiotypic antibodies directed against both the T cell receptors and IgG alloantibodies of that parental strain with specificity for alloantigens of the other parental strain. In order to prove identity between the anti-idiotypic antibodies against the B and T-cell antigen-binding molecules the following experiments were carried out; highly purified IgG from relevant alloantibody-containing serum in immunosorbent from could be shown to selectively remove both anti-idiotypic activities from the F1 antiserum. Further more, parental normal T lymphocytes could be shown capable of removing from the anti-idiotypic antisera all those antibodies that would cause agglutination of the relevant alloantibody-coated erythrocytes in the indirect agglutination assay. We would thus conclude that T and B lymphocytes reactive against a given antigenic determinant use receptors with antigen-binding areas coded for by the same variable gene subset(s).

Adsorption↗
PubMed1975

Inhibition of mixed leukocyte culture by anti-idiotypic antibodies.

(CBA x C57BL/6)F1 antisera prepared by injecting F1 mice with CBA T lymphocytes or CBA anti-C57BL/6 alloantibodies were tested for their ability to inhibit the mixed leukocyte culture (MLC) response. Presence of antiserum throughout the culture period in the absence of complement did not have any effect on the MLC response. Treatment of CBA responder cells with F1 antiserum and complement prior to the culture specifically inhibited the MLC response. Specificity of the suppression was ascertained; absorption of the F1 sera with F1 or C57BL/6 spleen cells did not remove the suppressive factor, whereas absorption with CBA spleen cells did so. F1 antiserum treatment left intact the response to third party alloantigens (DBA/2). Immunabsorbent columns with alloantibody of corresponding specificity removed the suppressive factor from anti-T cell sera as well as from antisera to alloantibody. The data suggest that the circulating alloantibody population contains molecules which share idiotypic determinants with surface receptors on T cells recognizing the same alloantigens.

Animals↗
PubMed1975

Idiotype positive T cells visualized by autoradiography and electron microscopy.

Anti-idiotypic antibodies were prepared in (CBA X C57BL/6)F1 hybrid mice by immunization either with CBA anti-C57BL/6 alloantiserum or with purified CBA thymus-processed lymphoid cells (T cells) Iodinated anti-mouse Ig or triple sandwich ferritin-labeling techniques served to visualize the reaction between idiotype and anti-idiotype. From 5 to 10% of purified CBA T cells appeared to carry receptors for C57BL/6 antigens. Heavily labeled cells had the morphology of small lymphocytes.

Animals↗
PubMed1975

Local graft-versus-host-reaction in mice specifically inhibited by anti-receptor antibodies.

The local graft-versus-host (GVH) reaction provoked by parental spleen cells in F1 mice was shown to be T-cell-dependent. GVH reactions were suppressed in F1 hybrid mice immunized with parental T lymphocytes of the same genotype, but not in F1 mice immunized with parental B cells. In some cases this immunity could be passively transferred by serum into normal F1 mice. The specific activity of such sera could be removed by absorption with either parental T or B cells. Some of the F1 antisera were specificlly cytotoxic for parental GVH-reactive lymphocytes.

Animals↗
PubMed1975

Shared idiotypic determinants on B and T lymphocytes reactive against the same antigenic determinants. IV. Isolation of two groups of naturally occurring, idiotypic molecules with specific antigen-binding activity in the serum and urine of normal rats.

In the rat the major histocompatibility locus antigens are determined by the Ag-B locus. In the present article evidence is presented that the sera and urine of normal adult rats contain naturally occurring antibody-like molcules with reactivity to allogeneic Ag-B antigens. Such molecules can be shown to contain both antigen-binding capacity for the relevant antigens and the idiotypic markers signifying such specific reactivity. The molecules could be shown to be composed of two groups of molecules, one around 7S IgG in size and the other around 35,000 in molecular weight. Only the smaller molecules were found in the urine. From other data we know that the 7S molecules are produced by B lymphocytes and the 35,000-molecular-weight molecules by T cells. Purified natural anti-Ag-B factors, when inoculated into rabbits or chickens, lead to the production of specific anti-idiotypic antibodies that will selectively inactivate rat T lymphocytes with the capacity to react against the relevant Ag-B antigens while leaving other reactivity intact. We thus conclude that the present system allows the purification of naturally occurring idiotypic B- and T-cell products with antigen-binding specificity for further biochemical and functional analysis.

Animals↗
PubMed1974

Cell-bound receptors for alloantigens on normal lymphocytes. II. Antialloantibody serum contains specific factors reacting with relevant immunocompetent T lymphocytes.

The rat popliteal node graft-vs.-host assay was shown to depend on the presence of parental T cells in the inoculum. Antialloantisera raised in F(1) hybrid rats against alloantibodies of one parent directed at transplantation antigens of the other parent displayed some or all of the following specific effects on parental T cells: They inhibited local GvH by purified T-cell suspensions; they blocked the capacity of GvH-reactive cells to adsorb onto fibroblast monolayers of the relevant genotype; together with complement, they killed GvH-reactive cells.

Animals↗
PubMed1974

Cell-bound receptors for alloantigens on normal lymphocytes. I. Characterization of receptor-carrying cells by the use of antibodies to alloantibodies.

Antialloantibodies were prepared in F(1) hybrid rats by immunization with alloantibodies from one parent raised against antigens of the other parent. The Ig fraction of such antialloantibodies was iodinated and used to investigate the nature of idiotype-carrying normal lymphoid cells. Lymphoid cell populations of the proper genotype fixed radioactive antialloantibody in proportion to their B-cell content. Activated T-cell populations, when depleted of B cells, did not fix significant amounts of radioactivity. Idiotype-carrying cells were sensitive to rabbit antirat Ig serum lysis and to antialloantiserum lysis, but not to rabbit anti-T-serum lysis. Also, the normal idiotype-containing B cell could be shown to have the expected antigen-binding specificity of its receptor. This was shown by the use of fibroblast cell monolayers that adsorbed specifically those cells capable of fixing the proper antialloantibody.

Animals↗
PubMed1972

Cellular receptors: binding of radioactively labeled anti-alloantiserum.

RAT ALLOANTISERA OF THE FOLLOWING SPECIFICITIES WERE OBTAINED AFTER SINGLE SKIN GRAFT REJECTION: DA anti-BN, BN anti-DA, DA anti-Lewis, and Lewis anti-DA. Anti-alloantibodies were raised by injecting the first two alloantisera into (DA x BN)F(1) hosts and the last two alloantisera into (DA x Lewis)F(1) hosts. In this manner, four anti-alloantisera were raised: anti-(DA anti-BN), anti-(BN anti-DA), anti-(DA anti-Lewis), and anti-(Lewis anti-DA). From each anti-alloantiserum a gamma globulin fraction was prepared and trace labeled with (125)I. The labeled anti-alloantibodies could be shown to be fixed preferentially by red cells coated with the alloantibody used to induce them. They were also preferentially fixed by normal rat lymphoid cells presumed to carry recognition structures similar to alloantibody. For instance, anti-(DA anti-BN) was fixed by BN red cells coated with DA anti-BN and by normal DA lymphoid cells presumed to carry RS(BN). Conversely, anti-(BN anti-DA) was fixed by DA red cells coated with BN anti-DA and by normal BN lymphoid cells, carrying RS(DA). Absorption and inhibition experiments showed that alloantibodies and normal lymphoid cells compete for the same labeled anti-alloantibodies. We conclude that cellular receptors and circulating alloantibody are antigenically similar: They have the same aliotype.

Animals↗
PubMedDate not supplied

Binding to leukocytes and induction of TNF alpha production involve different sites within the lipid-A region of LPS-like molecules.

The induction of TNF alpha synthesis in whole blood culture assay and isolated peripheral blood mononuclear cells was investigated, using LPS from Klebsiella pneumoniae and two water-soluble 34 kDa derivatives designed as acylpolygalactoside (APG) and EFA-APG, an APG molecule bearing two additional ester-linked fatty acids. Both APG and EFA-APG bind to monocytes by specific ligand receptor interaction but only EFA-APG could induce TNF alpha synthesis. It is concluded that ester-linked fatty acids are not involved in LPS binding to the cell surface, but play a critical role in the triggering of cellular responses.

Carbohydrate Sequence↗