Allergie und Immunologie最新文献

Different monospecific antisera against thiol-protein disulfide oxidoreductase (TPO, EC 1.8.4.2, protein-disulfide isomerase, EC 5.3.4.1) were raised in rabbits by immunization with purified human TPO and characterized by means of Laurell and immunoblot techniques. A competitive anti-TPO-EIA with insolubilized TPO has been used to determine this enzyme in cells and tissue homogenates. The assay shows a sensitivity of 1.2 ng/ml and a specificity of about 99%. The TPO content in relation to the total protein was found to be: in pancreas 0.65%, liver 0.45%, spleen 0.12%, placenta 0.16%, tonsils 0.06% and lymph nodes 0.03%.

Six lymphocyte antigens were detected with monoclonal antibodies in 0.2 ml capillary blood by a whole blood micromethod. After the lysis of erythrocytes of one drop of blood the remaining cells were layered on slides and were stained by indirect immunofluorescence. The relative and absolute counts of the lymphocyte subsets were of normal value and did not differ from the counts of flow cytometry and of slide and tube tests.

Mice treated intraperitoneally with zymosan showed a strong inhibition of the DTHR (delayed-type hypersensitivity reaction) against sheep erythrocytes (SE), ovalbumin (OA), and alloantigen. Furthermore, the rejection time of skin transplants was nearly doubled while antibody formation against SE was significantly enhanced. When a DTHR against OA and an antibody formation against SE were induced at the same time in the same animals, than the suppressive and stimulating effects cancelled each other. These results are discussed with regard to the sensitivity of lymphocyte subpopulations, which may be different if exposed to phagocytosis-induced oxygen radicals.

In this report, optimum conditions were determined for the production of immunoreactive fragments from a mouse monoclonal insulin antibody, and their immunochemical characterization is described. Stable fragments can be obtained in good yield from the purified IgG 1, first by cleavage with pepsin and then by reducing the disulfide bonds with cysteine and subsequent alkylation with iodoacetamide. F(ab')2 and Fab' fragments having molecular weights (Mr) of about 108,000 (108 K) and 55 K, respectively, were produced. Ligand binding assay as well as indirect immunofluorescence on mouse pancreas section demonstrated their reactivity with free and tissue-bound insulin antigen. These results provide methods for the production and characterization of defined fragments of insulin antibody useful in experiments where non-specific interactions mediated by the Fc portion of the whole immunoglobulin may occur.

Interleukin-1 mediates a broad spectrum of activities in the functional network of cytokines. In addition to its function as an inducer of the acute phase response IL-1 has many effects on hemopoiesis in normal and hematologically impaired organisms. This regulatory function is realized by its ability to stimulate the release of hematopoietic growth factors and by its recruiting property for cell cycles of different hemopoietic progenitors and stem cells. IL-1 acts synergistically with the colony-stimulating factors.

A group of children with atopic asthma (42) and the control group of normal children (30) were examined in order to determine the level of suppressor and helper T lymphocyte subpopulations in each group by using monoclonal antibodies. The asthmatic children were divided into two subgroups: one group received no therapy (30) and the other was treated with specific hyposensitization (12). Suppressor T lymphocytes were significantly lower in the subgroup of non-treated asthmatic children (p less than 0.01) and significantly higher (p less than 0.01) in the subgroup of children treated with immunotherapy. The stimulation index using mitogens was higher in the group of asthmatic children. The results suggest that the reduction and functional damage of suppressor T lymphocytes may have an influence on the pathogenesis of asthma and that immunotherapy may be beneficial in the treatment of atopic asthma in children.

Monoclonal antibodies against streptokinase as ligands were coupled to different matrices. Both CNBr-activated Sepharose and macroporous bead cellulose activated by carbonochloridate revealed as suitable for purification of streptokinase by affinity chromatography. Immunoadsorbents with a higher concentration of the coupled ligand were more effective than those with a lower. For streptokinase optimal conditions of binding and elution without negative influence on structure and activity were ascertained. A buffer with slight alkaline pH was successful for desorption. Using this method it was possible to obtain pure streptokinase from several streptokinase containing media.

Both newborns and elderly adults suffer from physiological immunodeficiency. The molecular mechanisms responsible for this immunodeficiency are currently investigated by many laboratories. The aim of our investigations was to answer the question wether these immunodeficiencies could be influenced by bovine and/or human diacetyl-splenopentin, two newly developed immunostimulatory peptides. The in vitro effects of these peptides were studied using the lymphocyte transformation test and the detection of the immunoglobulin production (IgG, IgM) by lymphocytes. Thymopentin was used as standard for these investigations. The age dependence of lymphocyte sensitivity was estimated using cells of the following groups of blood donors: newborns (cord blood); young donors (20-30 yr); old donors (over 70 yr). All peptides were shown to have the same effects. The stimulated lymphocyte proliferation (PHA, anti-CD3) was inhibited in young donors and further increased in old donors. There was no influence in the case of newborns. The biological activity of human diacetyl-splenopentin was shown to be higher in comparison with bovine diacetyl-splenopentin.

For coupling 25 mg of bovine IgG (BGG) were given to 5 ml volumes of packed bead cellulose activated by 5-norbornene-2.3-dicarboximido carbonochloridate and CNBr-activated Sepharose CL-4B, respectively. Thus, BGG-immunosorbents were obtained with 4.6 to 4.9 mg BGG/ml matrix. 5 ml volumes of packed NaIO4-oxidized Sepharose 6B coupled 27% from 25 mg of BGG, only. In this case, immunosorbents with 1.35 mg BGG/ml matrix were produced. All BGG-immunosorbents were chemically relatively stable. The use of these immunosorbents for affinity chromatography results in the isolation of one milligram of pure rabbit anti-BGG antibodies by means of about 4.6 mg of BGG coupled to the cellulose or the Sepharose-CL-4B matrices. On the other side, only 3.4 mg of BGG coupled to the NaIO4-activated Sepharose 6B were necessary in order to isolate one milligram of antibodies in an immunoelectrophoretically pure state.

Fc receptor mediated immune complex clearance function was measured in patients with systemic lupus erythematosus. Autologous erythrocytes were sensitized by human IgG anti-Rh(D) and used as immune complex model. An impaired Fc receptor function was demonstrable in 20 of 25 investigated patients. We have found a significant correlation between the seriousness of the defect and the step of the immunologic activity of the patients, but not between the impaired Fc receptor function and clinical activity and renal manifestation in our SLE patients. Further studies are necessary to determine the relevance of this phenomenon and to clear the possibility of therapeutical influence.