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Biomedical subjects

H Frischknecht

Publications and source records attributed to H Frischknecht.

11 recordsLinked to original sources

HB Hinwil or beta 38(C4)Thr-->Asn: a new beta chain variant detected in a Swiss family.

This paper reports a new hemoglobin variant which was identified while investigating the cause of a mild erythrocytosis. The abnormal beta-globin chain was detected by reversed phase chromatography. Mutation mapping of the beta-globin gene by polymerase chain reaction and denaturing gradient gel electrophoresis followed by sequence analysis revealed a C-->A transversion at codon 38, predicting a Thr-->Asn substitution. Tryptic peptide mapping by liquid chromatography electrospray mass spectrometry, followed by conventional Edman peptide sequence analysis, confirmed the predicted amino acid substitution. In contrast to the only other known mutation at codon 38, Hb Hazebrouck (Thr-->Pro), this hemoglobin is stable and shows elevated oxygen affinity.

Amino Acid Sequence

Application of dynamic capillary isoelectric focusing to the analysis of human hemoglobin variants.

Capillary isoelectric focusing (CIEF) with electro-osmotic zone displacement of normal and pathological hemoglobins (Hb) is reported. CIEF is performed in untreated, open-tubular, fused-silica capillaries of 75 microns internal diameter using methylcellulose for dynamic column conditioning. After direct injection of hemolysates mixed with carrier ampholytes, high resolution separation of Hb variants, including Hb A1c, A, F, D, S, E and A2, is obtained, this permitting unambiguous characterization of Hb patterns of normal adults, newborns, patients with diabetes, different hemoglobinopathies and thalassemia syndromes. Qualitatively, the CIEF data compare well with those obtained by gel isoelectric focusing and high-performance liquid chromatography. CIEF is demonstrated to be a simple, rapid and fully instrumental approach to Hb analysis. Run times of less than 20 min make CIEF an attractive method for routine Hb investigations and screening programs.

Adult

Binding of purified, soluble major histocompatibility complex polypeptide chains onto isolated T-cell receptors. I. Reactivity against allo- and self-determinants.

In this study, we tried to get information about the fine antigen-binding ability of purified, soluble, idiotype-positive T-cell receptor molecules. Lewis anti-DA T-cell receptors were purified from normal Lewis serum by the use of anti-idiotypic immunosorbent and sodium dodecyl sulfate-polyacrylamide gel, and were coupled to cyanogen bromide-activated Sepharose 4B. In parallel, Lewis anti-DA, Lewis anti-BN, and DA anti-Lewis alloantibody immunosorbents were prepared. The major Ag-B chain (44,000 daltons) and the two polypeptide chains (34,000 and 27,000 daltons) of Ia were purified from Lewis, DA, and BN lymphocytes and absorbent on the above-mentioned immunosorbents. We found that the major Ag-B chain as well as the two Ia chains were bound to the alloantibody columns if they were derived from the corresponding allogeneic strain. No retaining ability for self-major histocompatibility complex (MHC) or third-party MHC chains was noted with the alloantibody immunosorbents. When using immunosorbents made up of idiotypic T-cell receptors, only two MHC polypeptides of the relevant allo-MHC type were retained, namely, the Ag-B and the heavy Ia chains. No detectable activity was observed when testing the same column for reactivity against third-party MHC polypeptide chains. However, the Lewis anti-DA T-cell receptors could be shown to display weak, but significant, reactivity toward one Lewis MHC polypeptide chain, that is, the heavy chain of Ia type.

Animals

Idiotypic determinants on T-cell subpopulations.

Killer T cells with specificity for major histocompatibility antigens have been shown in mice and rats to display idiotypic receptors allowing the lysis of such cells at the effector phase by anti-idiotypic antibodies and complement. A comparison was made between idiotypes displayed by Lyt-1-2+3+ and Lyt-1+2-3- T blasts, generated in the same mixed leucocyte culture (MLC), across an entire H-2 locus barrier. This was done by absorption of anti-idiotypic antibodies with respective T blasts, followed by estimation of the ability of the absorbed antiserum to inhibit MLC or killer T-cell function. Further, the capacity of Lyt-purified, MLC-generated T blasts to provoke specific unresponsiveness via anti-idiotypic immunity in syngeneic recipients was analyzed. Taken together, the results demonstrate that Lyt-1+2-3- T blasts responsible for the major part of MLC proliferation have distincly different idiotypes from those on the Lyt 1-2+3+ killer T cells. That the idiotypes on the killer T-cell presursors can serve as triggering sites for induction of effector T-cell function was then suggested by experiments with Lyt-1-2+3+-purified, normal T cells as precursor cells in vitro. The fact that autoanti-idiotypic antibodies may circumvent the need for helper Lyt-1+2-3- T cells in the generation of allospecific killer T cells indicates that the former cells may normally function partly via such anti-idiotypic reactions.

Animals

Alloantigens derived from stimulator cells and bound onto MLC-activated rat T lymphoblasts.

Rat T lymphocytes were activated in MLC (mixed leucocyte culture) against allogeneic stimulator cells. A few percent of such T blasts could be shown to carry IgM on their surface when a mixture of T and B lymphocytes was used as responder cells. The immunoglobulin was absent when purified T lymphocytes served as responders. A large proportion of MLC T lymphoblasts could be shown to carry stimulator alloantigens on their surface. These antigens were demonstrated (a) by indirect immunofluorescence using anti-stimulator alloantibody and FITC (fluorescein isothiocyanate Isomer I)-labelled F(ab)2 fragments of rabbit anti-rat Ig; (b) by autoradiography after internal labelling of stimulator cells, which showed the transfer of label from stimulator to responder cells. For biochemical analysis T blasts were restimulated with internally labelled allogeneic or syngeneic stimulator cells, and bound radiolabelled stimulator alloantigens were analysed on SDS-PAGE (sodium lauryl sulphate polyacrylamide gel electrophoresis). The results suggest that intact SD antigens with a molecular weight of 45,000 daltons and LD determinants with a molecular weight of 34,000 daltons derived from stimulator cells are found on MLC T blasts. No evidence of binding of the 27,000-dalton Ia chain to T blasts was found. Possible arrangements of these molecules on the responder blasts is discussed.

Animals

Representation of heavy but not light chain Ig idiotypes on T cell receptors for alloantigens.

We have analyzed idiotypic determinants on B and T lymphocytes reactive against the same antigenic determinants by the use of different anti-idiotypic antibodies. Such antisera were produced in (Lewis X DA) F1 rats against Lewis anti-DA alloantibodies (= B cell product) and Lewis T lymphocyte receptors with the same specificity. We found that B lymphocytes bear unique idiotypic determinants which are not present on the corresponding T lymphocytes. T cell unique (not shared by B lymphocytes) idiotypes were so far not detected. T cells idiotypic determinants which are present on heavy but not light chains of the corresponding alloantibodies.

Animals

Induction of specific transplantation immune reactions using anti-idiotypic antibodies.

B and T lymphocytes with reactivity against major histocompatibility antigens are known to express this immune potential via a display on the outer surface of antigen-specific, idiotypic receptors. Here, we show that anti-idiotypic antibodies directed against such receptors may serve as specific triggering agents of the idiotype-positive lymphocytes in the physical absence of foreign histocompatibility antigens. This was shown in vitro using normal or immune spleen T cells where anti-idiotypic antibodies would lead to the selective proliferation and development of antigen-specific cytolytic T cells as determined by short-time 51Cr release assays. Furthermore, purified anti-idiotypic antibodies in adjuvant administered in vivo to normal syngeneic animals could be shown to lead to production of high titers of specific alloantibodies. The present experiments were in most cases carried out using auto-anti-idiotypic antibodies as triggering agents. The present results thus lend further support to the concept that idiotype-anti-idiotype reactions may be normal parts of conventional immune processes with either stimulatory or inhibitory consequences, depending upon the prevailing conditions.

Animals

Some studies on idiotypes and anti-idiotypic reactions and receptors in anti-allo-MHC T-cell immunity.

The fine specificity of immunocompetent T lymphocytes is reflected on their outer surface by their display of idiotype-positive receptor molecules. Interference with these structures using antiidiotypic reagents may trigger or eliminate the cell depending on the external conditions. When analyzing the distribution pattern of idiotypes on murine allo-MHC activated T blasts it was found that proliferating "helper" T and killer T cells display non-overlapping sets of idiotypes. Furthermore, it was found that antiidiotypic antibodies in vitro may mimic the allo-MHC antigens in the T-cell triggering capacity and may indeed in some systems be superior to allogeneic stimulator cells in this regard. This capacity of antiidiotypic antibodies to functionally mimic and replace allo-MHC structures in T-cell triggering ability is discussed in relation to self-MHC reactivity and biochemical and functional similarities between Ig molecules and MHC structures.

Animals