Biomedical subjects
H Ambrosius
Publications and source records attributed to H Ambrosius.
Sepharose gels as antigenic matrices for specific binding of anti-galactosyl antibodies.
From 100 ml of pooled normal serum taken from different species (man, cow, rabbit, chicken, duck, carp) 0.5 to 4.0 mg of high and low molecular weight anti-galactosyl antibodies could be isolated by affinity-chromatography on non-ligand-attached Sepharose 4B. The antibodies bound to the agarose matrix were eluted with 0.5 M D-galactose containing buffers. Using species-specific anti-immunoglobulin sera in immunoelectrophoresis, the immunoglobulin character of the protein fractions prepared was analyzed. In addition, the H and L chain structure of the anti-galactosyl antibodies was checked by SDS-PAGE.
[Enzyme immunoassay for the determination of IgE antibodies specific for grass pollen allergens using monoclonal antibodies of the BL-IgE series. 1. Methodologic study and optimization of the test system].
The development and optimization of an enzyme immunoassay for the determination of allergen specific antibodies of the IgE-class using a standardized grass pollen allergen and the monoclonal antibody BL-IgE 9 is described.
[Enzyme immunoassay for the determination of IgE antibodies specific to grass pollen allergens using monoclonal antibodies of the BL-IgE series. 2. Performance and results of a collaborative study to evaluate the test system].
This paper describes performance and results of a collaborative study in which the solid-phase enzyme immunoassay was tested in order to determine specific IgE under practical conditions. All titres of specific IgE can be differentiated in a reproducible way with high significance.
[Transplantation antigen specific rosette test and its inhibition with autologous serum--a new immunologic method for predicting rejection crisis following allogeneic organ transplantation].
A combination of transplantation-antigen-specific rosette test with inhibition of rosette formation, using autologous serum, was used in postoperative follow-up checks on patients who had undergone allogeneic kidney transplantation and proved successful in early diagnosis or prediction of rejection episodes even before clinical manifestation. The numbers of antigen-detecting and antigen-binding cells differed considerably from each other, depending on patients who had undergone kidney transplantation. Almost complete inhibition of antigen detection was generally achieved by addition of patient serum. If sensitization to alloantigen in excess of the limit value (30 RBZ/10(3) KHZ) is accompanied by rosette formation with autologous serum addition in excess of about 18 RBZ/10(3) KHZ, clinically relevant rejection can be expected to occur after another two or three days and will have to be treated by immunosuppressives. Such partial or even complete elimination of rosette inhibition is usually only of short length (one day) and has rarely been observed to last longer. No rejection calling for treatment will develop, on the other hand, as long as blocking serum factors provide for high-stability inhibition of any bond of transplantation antigen (rosette formation). Constantly low rosette formation with serum addition and without was exhibited by control persons (blood donors, patients with cerebrocranial trauma) in daily examinations. Limit values of 18 or 30 RBZ/10(3) KHZ were not surpassed.
The influence of environmental temperature on the lymphocyte populations in carp (Cyprinus carpio L.).
The influence of environmental temperature upon the lymphocyte populations in carp (Cyprinus carpio L.) has been studied by immunoperoxidase labeling using different antisera. In carps kept in a warm and a cool environment, respectively, rabbit anti-carp IgM as well as rabbit anti-carp Fc antiserum showed different percentages of Ig-positive lymphocytes, whereas the percentage remained unaffected by temperature after the use of rabbit anti-carp Fab antiserum with a specificity against L-chains. It is clear from these observations that the "B" lymphocytes of carp are not temperature-dependent, while the "T" lymphocytes are extremely temperature-dependent, decreasing at low temperatures and increasing at elevated temperatures. The possible causes of the variation with temperature in the numerical ratios of lymphocyte populations and their importance for immunoreactivity are discussed.
[Detection of a TAA-specific sensitization in patients with metastatic breast cancer following breast removal and upper and lower half-body irradiation].
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[Principles of immune regulation].
The regulation principles effective in an immune response are demonstrated in form of a review. They comprise the regulation by lymphocyte-subpopulations, the feedback by made up antibodies as well as the idiotype-anti-idiotype regulation. Conclusions are drawn for the clinic as well as the necessary research disciplines.
Evolution of low molecular weight immunoglobulins. V. Degree of antigenic relationship between the 7S immunoglobulins of mammals, birds, and lower vertebrates to the turkey IgY.
There are close antigenic relations between the turkey IgY--the dominant 7S Ig of serum--and the corresponding IgY-like Igs of chicken, duck, goose, tortoise, and frog as well as serum or colostral IgA of man and swine. The RIA-systems used for the determination of immunological cross-reactivity among the Ig types consisted of 125-I-labelled turkey IgY and class-specific rabbit anti-turkey IgY antibodies. The results stand in a good agreement with our findings demonstrating strong antigenic relationships between various serum or colostral Igs of IgY and IgA types, respectively, to the chicken IgY. Therefore, the IgY-like Igs seem to be the precursors for the IgA class of mammals.
[Comparison of intrasplenic immunization with other types of antigen application].
Mice have been immunized intrasplenic (i.l.), i.p., or i.v. with different doses of bovine gammaglobulin (RGG). The number of antigen reactive cells (ARZ) in the spleen was measured by immune rosette technique. Antibodies were detected by passive hemagglutination. The i.l. immunization induced no higher numbers of ARZ than the other ways of antigen application, but the maximum arose earlier, already at the third day. The most ARZ were B lymphocytes. Very low doses of antigen induced antibodies only with i.l. immunization. Most antibodies at the maximum of the primary immune response were of IgM class. A typical secondary response could not be elicited following i.l. priming. Our investigations don't refer to advantages of i.l. immunization for hybridoma techniques.
[Immunologic characterization of the rejection of the allogenic transplanted kidney in pigs with and without immunosuppression].
After allogenic renal transplantation in pigs kinetics of the immunological reactivity have been investigated in order to diagnose rejection crisis. In a first series of 26 transplants without any immunosuppression we secured the diagnosis of rejection in 95% of cases of acute or chronic rejection by a combination of antigen-specific rosette test, inhibition of this parameter by autologous serum, complement dependent cytotoxicity and antibody dependent cell-mediated cytotoxicity. The antigen-specific rosette test reacts very early, 2-3 days before a bad clinical feeling in the course of acute rejection or 5-14 days by chronic rejection. Under the therapy with prednisolone and azathioprine we could not reach any prolongation of survival by arbitrary donor-recipient-selection. Under immunosuppression antibodies have never been observed by acute rejection in CDC and ADCC. In every case of acute rejection an increasing rosette formation of specific antigen binding cells appeared 2-3 days before an increasing of serum creatinine. In the case of other complications like infection or thrombosis this test is not reacting. However, by thrombosis of arteria or vena renalis there can be an increased specific rosette formation either independent of this process by beginning of immune reaction or by causal connection of both processes (necrotizing thrombotic rejection). So we have given the evidence, that the antigen-specific rosette test on a high level meets all requirements of a specific diagnosis of rejection under immunosuppression in pig.
Comparative immunology as a research tool.
Only very few immunological publications are based on investigations using non-mammalian vertebrates as experimental animals. Nevertheless, some important immunological problems, such as dichotomy of T and B lymphocytes or immune tolerance, were first discovered or even solved using such species. The usefulness of comparative investigations with species of different vertebrate classes is shown, referring to our own investigations of organ transplantations in the 1950s and of the regulation of the IgM antibody response in the last years.
Radioimmunochemical studies on 7.8S and 5.7S duck immunoglobulins in comparison with Fab and Fc fragments of chicken IgY.
In ducks two forms of low molecular weight Ig's +), 7.8S and 5.7S, have been described. To compare their antigenicity with Fab and Fc fragments of chicken IgY a double-antibody RIA technique was used in which the binding of 125I-chicken IgY or 125I-chicken IgY (Fc) to rabbit and carp anti-chicken IgY (Fab) and anti-chicken IgY (Fc) antibodies, respectively, was inhibited by duck 7.8S and 5.7S Ig's. Chicken IgY and duck 7.8S Ig ( IgY -like protein) are highly cross-reactive with respect to their Fab as well as Fc part determinants. On the other hand, the duck 5.7S Ig shows nearly identical determinants to the Fab fragments of 7.8S Ig, but a Fc part is lacking. Therefore, we conclude that the 5.7S Ig molecule in ducks doesn't represent a separate Ig class. It consists of the same principal type of heavy chains as the 7.8S Ig, but the H chain of the 5.7S Ig lacks very likely the last two homologous constant regions.
Evolution of low molecular weight immunoglobulins--IV. IgY-like immunoglobulins of birds, reptiles and amphibians, precursors of mammalian IgA.
Radioimmunochemical studies on the comparison of the immunological cross-reactivity between the 7-S Igs of birds, reptiles and amphibians (IgY-like Igs) and the IgA of mammals (man and pig) using 125I-chicken IgY and anti-chicken IgY(Fc) or anti-chicken IgY(H) antibodies from rabbits and carp for the detection led to the conclusion that there are close antigenic relationships between them. Therefore, the IgY-like Igs seem to be the precursors for the IgA class of mammals. From that, we give a phylogenetic tree of Igs in accordance with the evolutionary development of vertebrates.
Conformational changes induced by hapten in murine monoclonal antibodies to dinitrophenyl groups--the analysis by temperature-perturbation spectroscopy.
Two samples of murine monoclonal antibodies to dinitrophenyl groups were studied by difference thermal perturbation spectroscopy with particular attention to changes in the amount of perturbed chromophores induced in antibodies as a result of hapten binding (epsilon-2,4-dinitrophenyl-L-lysine). Despite the fact that both antibody samples belong to immunoglobulin G1 and have the same type of light chain, kappa, they were found to differ significantly in the number of the chromophores perturbed by temperature. The binding of hapten decreases the perturbation of chromophores only in the sample with the less rigid structure, as regards thermal perturbation. These data provide evidence that differences in the rigidity of the structure of variable domains affect the extent of conformational changes induced in the antibodies due to the interaction with an antigen.
[Methodological studies on the hemolytic anti-bovine gamma globulin plaque technic].
Sheep red blood cells (SRBC) coated with specific bovine antibodies are suitable to detect anti-bovine gammaglobulin (BGG)-antibody-forming cells with the hemolytic plaque technique. The number of the definable anti-BGG-plaque-forming cells is dependent on the antibody density per SRBC. The developed plaques are specific for BGG. They are to inhibit with BGG only not with other proteins. The cellular kinetics of the anti-BGG response is to follow up in the spleen and poplitea lymph nodes of the immunized mice with this modification of the hemolytic plaque technique. The results are discussed.
[The importance of active rosette-forming cells and specific antigen-binding cells in the diagnosis of rejection after allogeneic orthotopic kidney transplantation].
After allogeneic renal transplantation in 26 female pigs the kinetics of the immunological reactivity against the graft have been investigated with a specific test, the antigen specific rosette test and an unspecific test, the active E-rosette test. In acute and chronic rejection the level of antigen specific rosette forming cells reflects very well the clinical signs of rejection. Because of early occurrence of immunological reactivity in the case of frequent (daily) testing this test is suitable for a relative sure prediction of rejection episodes. In spite of their relative good correlations to the increase of serum creatinine, posttransplant increase and decrease of active rosette forming cells are not always in an uniform temporal relation to the rejection process. For that reason this test alone is not suitable for diagnosis of rejection. However, high preoperative values (greater than or equal to 18% active T cells) refer to an increased readiness to immunological reactivity of recipient against the graft and reflect contraindication to transplantation. If active rosette forming cells (greater than or equal to 18%) and antigen specific rosette forming cells show maxima at the same time, than follow in high significance strong cell mediated rejections. On this way the active E-rosette test completes the values of the antigen specific rosette test.
[A further confirmation of the high idiotypic specificity of carp anti-human myeloma protein sera].
Previous investigations about the specificity of carp anti-idiotypic sera could be confirmed by comparison with corresponding antisera of guinea pigs. 3 carp antihuman myeloma-protein sera rendered as idiotypic specific after an appropriate absorption were tested in the passive hemagglutination inhibition test. They showed no difference to the parallel tested guinea pig antisera. It can be assumed that between fish antibodies and antibodies of mammals no functional difference exists in respect of antigen binding. On the other hand mammals exhibit a higher extent of structural antibody heterogeneity.