IgE-reactivity to major Japanese cedar (Cryptomeria japonica) pollen allergens (Cry j 1 and Cry j 2) by ELISA in dogs with atopic dermatitis.

The present study investigated IgE-reactivity to two major Japanese cedar (Cryptomeria japonica, C. japonica) pollen allergens (Cry j 1 and Cry j 2) in dogs with atopic dermatitis by use of a fluorometric ELISA. The serum samples from 27 dogs that showed IgE-sensitivity to crude C. japonica pollen allergen by ELISA were tested for specific IgE to the two major allergens. All 27 dogs had anti-Cry j 1 IgE, and 10 (37%) had anti-Cry j 2 IgE. Inhibition of binding of dog specific IgE to crude C. japonica pollen allergen was carried out by addition of Cry j 1. When serum samples containing anti-Cry j 1 IgE but no anti-Cry j 2 IgE were incubated with Cry j 1, specific IgE binding to crude C. japonica pollen allergen was almost abolished. These findings suggest that Cry j 1 is a major allergen in dogs.

Anti-IgE autoantibodies mistaken for specific IgG.

During isolation of specific IgE by affinity chromatography, we obtained IgG acting as "specific IgG" in dot immunoassay and RAST. Further analysis of this fraction revealed that this specific IgG was indeed an immune complex consisting of IgG anti-IgE autoantibodies and specific IgE that falsely appeared as specific IgG in the immunoassays. After disruption of complexes, more IgE became accessible, resulting in increased counts per minute in the RAST. Thus, anti-IgE autoantibodies may hinder determination of specific IgE in in vitro immunoassays, since they hide specific IgE immunologically. To demonstrate the isotope specificity of such immune complex-derived IgG, the purified anti-IgE autoantibodies were radiolabeled and used as detection antibodies in immunoblotting. These autoantibodies recognized different IgE preparations, including a recombinant IgE peptide.

Development of a serum-free defined culture medium for lymphoblast transformation tests of mouse spleen and thymus cells.

By analysis of thymidine uptake during the first 4 h of incubation and by examining the responsiveness of unfractionated mouse spleen cells upon mitogenic or allogeneic stimulation, some serum factors of major importance for in vitro cultivation of lymphocytes have been examined. Albumin and l-alanine are essential for the maximal preservation of the in vivo-initiated lymphocyte activity during the first hours of incubation in vitro. Transferrin plays a major role in the lymphocyte proliferation, induced by mitogens in vitro, and, finally, zinc and selenium exert a clear enhancing effect on the response to an allogeneic stimulation. The AATSZ medium (RPMI 1640 enriched with l-alanine, albumin, transferrin, zinc chloride, and sodium selenite) enables a proliferation of the same magnitude as or higher than FCS medium. The kinetics are the same, and the cell viability is comparable, but standardization is much simpler with AATSZ. This is primarily because FCS binds some mitogens and contains inhibitors. Consequently, the standardization of such a culture system is dependent on variations from serum batch to serum batch. On the other hand, the current composition of the AATSZ medium promotes the sticking capacity of T lymphocytes and does not support growth of all lymphoid cell lines. Consequently, this defined medium, although not yet suitable as optimal medium for all lymphocyte functions, can advantageously be used for short-term studies of most murine lymphocyte functional and cooperation studies in vitro.

Equine leukocyte antigen system. I. Serological studies.

Lymphocytotoxic alloantibodies have been recognized in primiparous mare sera and colostra using the two-step microcytotoxicity test. The antigens detected on the leukocyte membrane do not occur on the erythrocytes. After testing on a cell panel from unrelated horses, absorptions with leukocytes were carried out. The subsequent retesting of the serum reagents on a cell panel including cells from 24 families rendered possible a grouping of preliminary monospecific reagents characterizing 18 antigen specificities on the cell membrane.

[Comparative study on the immunogenicity of plasma substitutes with gelatin base in animal experiments]. / Vergleichende Studie über die Immunogenität von Plasmaersatzlösungen auf Gelatinebasis im Tierexperiment.

The immune response of rabbits and guinea pigs to gelatin and to three commercial plasma substitutes based on gelatin (oxypolygelatin [OPG], di-isocyanate cross-linked gelatin [DCG], modified fluid gelatin [MFG]) has been reevaluated. Passive hemagglutination, passive cutaneous anaphylaxis and active anaphylaxis have been used to detect the response of animals immunized in complete Fruend's adjuvant or in aluminium hydroxide. Marked immune responses were observed with DCG and MFG which were to a large extent specific for the immunizing antigen (i.e. the corresponding chemically modified gelatin). Gelatin and OPG induced weak responses. In contrast to DCG and MFG in systemic anaphylactic shock experiments no anaphylactic shock could be elicited with gelatin and OPG. A limited series of immunizations in guinea pigs of various strains demonstrated that responses to weak immunogens, such as modified gelatins, are markedly influenced by genetic factors.