Search PubMedSearch

Biomedical subjects

H H Sedlacek

Publications and source records attributed to H H Sedlacek.

18 recordsLinked to original sources

Expression of GD2-epitopes in human intracranial tumors and normal brain.

Two monoclonal antibodies (mabs) were raised against ganglioside GD2 (Gtri2) and tested on human intracranial tumors and normal brain by immunohistochemical methods both in frozen and paraffin embedded sections. In normal brain structures, astrocytes were visualized with both mabs (BW 625 and BW 704) almost exclusively in the subventricular and subpial layer. A minor amount of myelin sheaths in well defined localisation was only recognized in frozen sections. Consequently in astrocytic tumors of different grades of malignancy (WHO I-IV), astrocytes were depicted in their characteristic shape and arrangement around vessels. In addition, staining was observed in meningiomas and schwannomas, but not in pituitary adenomas or metastatic carcinomas. In meningioma und schwannoma the staining was restricted to the cellular periphery and was again present in frozen section material and absent in paraffin embedded tissue. In astrocytes, reactive and neoplastic, obviously fibrous processes and cytoplasm were distinctly stained both in frozen and paraffin embedded sections. It is concluded that some neuroectodermal derived cells as well as myelin of defined localisation express GD2 on the membrane surfaces and in the cytoplasm. The latter may primarily be the case in fibrous astrocytes, which were stained in reactive and pathologic conditions. The reaction can be used as diagnostic tool in astrocytic tumors; its positive therapeutic significance is hampered by the fact that (1) not all cells in heterogeneous tumor populations express the epitope and (2) there are normal structures which do so.

Antibodies, Monoclonal

[Henoch-Schoenlein purpura (author's transl)].

Routine EEG investigations and observance of discrete neurological and psychological symptoms in 13 children in the acute phase of Henoch-Schoenlein purpura showed that involvement of the central nervous system in this disease is the rule rather than the exception. Capillary resistance was reduced in 51 out of 76 investigated children. On the other hand a reduction in factor XIII activity was much less commonly found (n =6). Immune complex determination in 28 children, together with antibody studies, showed that in 13 of them the Henoch-Schoenlein purpura was triggered off by an influenza-A-virus infection of the upper respiratory tract. Two thirds of the patients had markedly raised levels of total serum complement which fell weeks within several.

Adolescent

A new method for fluorescence immunoassay using plane surface solid phases (FIAPS).

A method distinguished by high sensitivity, low non-specific binding, easy handling, and broad applicability with respect to various antigens is described. Films of polymethyl-methacrylate with plane surfaces were selected as solid phase for adhesive or covalent binding of different antigens (DNA, histone, human, rabbit or goat immunoglobulins). Proteins were covalently bound to the films by the azide method (Orth and Brummer, 1972). Polymethylmethacrylate films thus coated had a negligible autofluorescence and gave minimal non-specific binding of protein. Coated films were used for specificity control of FITC-labeled antibody preparations and in the double antibody and sandwich techniques for detection of antibodies or antigens in sera from man, rabbit and goat. FITC-conjugated hyperimmune antibody, in some cases purified by immunoadsorption was used as second antibody in indirect techniques. The amount of fluorescent-labeled antibody bound per unit of surface area of film was measured by incident light with a Zeiss-Axiomat fluorescence microscope equipped for fluorescence photometry and an uranyl acetate glass plate was used as a standard. The technique appears superior to present methods of quantitative immunofluorescence analysis.

Animals

Antigen-specific detection of soluble immune complexes by a solid phase specific antibody system.

An antigen-specific immune complex assay based on the following principle has been developed: xenogeneic, antigen-specific antibodies are attached to a solid phase. During first incubation with patient's serum, immune complexes in antigen excess are bound to the xenogeneic antibody by their free antigenic determinants. In a second incubation a labeled antibody, specific for human immunoglobulins, is combined with the antibody part of the immune complex. Quantitation of the label allows the determination of the immune complexes. The principle of the method has been varied using artificial soluble immune complexes of tetanus toxoid and human anti-tetanus toxoid antibody, and immune complexes prepared by mixing patient sera containing either HBsAg or anti-HBsAg antibodies. The reliability of the results is shown by their coefficient of variation (2.5%). With the method described soluble immune complexes predominantly in slight antigen excess, which are thought to be responsible for development of immune complex disease, have been detected.

Antibody Specificity

Antigen-specific detection of HBsAG-containing immune complexes in the course of hepatitis B virus infection.

In recent studies extrahepatic manifestations of viral hepatitis have been recognized as immune complex diseases. Hepatitis B surface antigen (HBsAg) has been successfully identified in immune complexes, but the pathogenic role of HBsAg-containing immune complexes (IC) remains questionable. The subject of the present study was the antigen-specific determination of IC in the course of hepatitis B virus infection using a new HBsAg-specific IC test (Pernice & Sedlacek, 1978). This test is based on the following principle: rabbit anti-HBs-coated polystyrole test tubes are incubated with the IC-containing test sample. The HBsAg-containing IC bind to the solid phase by their free antigenic determinants. There they can be quantified using a peroxidase-labelled anti-human IgG antibody. A good correlation was found between the level of HBsAg-containing immune complexes and the clinical state of six patients in a follow-up study. IC could be detected simultaneously with HBsAg and either decreased or disappeared before the occurrence of free anti-HBs. In the sera of an additional twenty-eight patient suffering from chronic active hepatitis, HBsAg-containing immune complexes were detected in 85% of cases. One patient suffering from polyarteritis nodosa was also positive. Occasionally, extremely high levels of IC were found in the course of these diseases.

Antigen-Antibody Complex

[Baby hamster kidney cells as antigen for demonstration of antinuclear antibodies (author's transl)].

Baby hamster kidney cells fixed in acetone on glass slides were used as antigen for demonstration of antinuclear antibodies. Where certain storage conditions were observed (drying agent, 4 degrees C) they have kept for 12 months up to now. As regards specificity, sensitivity, reproducibility, and differentiation of fluorescent types the baby hamster kidney cell test appears superior to other immunofluorescence methods used (chicken erythrocytes, rat liver sections, and crithidiae). These results were obtained in 73 sera from patients with disseminated lupus erythematodes, drug-induced lupus erythematodes, discoid erythematodes, allergic vasculitis, progressive scleroderma, dermatomyositis, and 36 control sera.

Antibodies, Antinuclear

Effect of Vibrio cholerae neuraminidase on the phagocytosis of E. coli by macrophages in vivo and in vitro.

The mechanisms by which Vibrio cholerae neuraminidase VCN acts as an adjuvant is unclear. We have studied the effect of VCN on the phagocyotsis of E. coli 0111 K58 by macrophages in vivo and in vitro. (1) Using and in vivo intraperitoneal clearance test we have shown that prior injection of VCN (50 units/mouse) into the peritoneal cavity of mice enhances the clearance of the bacteria as efficiently as an optimal amount of opsonizing antiserum. We have also shown that the enhancement of intraperitoneal clearance by VCN is due to phagocytosis and intracellular killing of the organisms by macrophages. (2) VCN does not enhance the i.v. clearance of E. coli o111 K58. (3) VCN promotes the attachment of the organisms to and phagocytosis by peritoneal macrophages in vitro. Data are provided which suggest that the promotion of attachment and phagocytosis by VCN cannot be explained by a mere increase of passive absorption of the bacteria to the cell wall.

Animals

[Tissue distribution of native and enzyme-treated human immunoglobulin (author's transl)].

The tissue distribution of intravenously injected native and enzyme-treated human gammaglobulin has been investigated in the small intestine, kidney, striated muscle and the salivary gland of the mouse. Peroxidase-as well as FITC-labeled antibodies to human gammaglobulin were used to locate the injected antibody preparation. With this assay, the enzyme-treated immunoglobulin was found intracellularly, whereas the intact molecule seemed not to penetrate the cell wall and could mostly be found intra-and perivascularly. Since highly purified tetanus toxoid was fixed by thin sections of the intestine tissue containing intracellularly located enzyme-treated gammaglobulin it is concluded that the antibodies penetrating the cell wall retain their biological activity.

Animals

Neuraminidase and tumor immunotherapy.

Preliminary results of first clinical studies with the enzyme neuraminidase call attention to a new kind of cancer treatment. This promising approach to tumor immunotherapy was entered into the clinical phase as a consequence of successful experimental studies in tumor-bearing mice, rats and dogs. In this review, the presently known and essential results of experimental and clinical studies on tumor immunotherapy by means of neuraminidase are presented as well as some necessary and critical considerations in this context. Moreover, out of a broad variety of results of biochemical and biological in vitro studies, it was attempted to select the more essential knowledge which could contribute to a better understanding of the still rather unclear in vivo mode of action of the enzyme neuraminidase. In a first brief paragraph (1.0), the biochemically characteristic data of the enzyme neuraminidase is presented. In the second section (2.0), the basic knowledge about the effects of neuraminidase on cell behavior is rather amply contained. Here, on the one hand, the biophysical and biochemical alterations are mentioned, the so-called ""unmasking'' effects are reconsidered and, on the other hand, the effects on the immunologically responding cell are discussed. In a third section (3.0), the diverse findings from animal experiments using neuraminidase-treated tumor cells are confronted, whereby tumor transplantation experiments and tumor therapy experiments are dealt with separately. The last section (4.0) reports about the first clinical studies with neuraminidase-treated autologous as well as homologous tumor cells, which partly brought about rather surprising and astonishing success. On the basis of recent findings by the study group of the authors, the more prior and sometimes discrepant results of various groups are critically considered. The problems of alteration of antigenicity and of other properties of cells through splitting off membrane-bound neuraminic acid, the facts of adjuvanticity of neuraminidase itself, the relation of successful therapy to dose dependency as well as the relation of undesirable methods for tumor mass reduction to the immunological responsiveness of the tumor bearer were especially looked into.

Animals

Regression of spontaneous mammary tumors in dogs after injection of neuraminidase-treated tumor cells.

The effect on tumor progression produced by the injection of VCN-treated tumor cells in dogs with spontaneous mammary tumors was investigated. Untreated dogs of different races and different ages with at least two palpable spontaneous mammary tumors were selected. One of the tumors was left in the animal for further clinical examination whereas the other tumor(s) was (were) excised for preparation of a single-cell suspension by mechanical disintegration and enzymatic digestion with collagenase and trypsin. (1) In the first group, each animal was infected with 2 times 10-7 similarly prepared autologous, mitomycin-treated tumor cells; in 8 out of 12 dogs of this group the tumors progressed while so far 1 dog has died of metastasis. (2) In the second group, each animal received the same number of 2 times 10-7 tumor cells, which were mitomycin- and VCN-treated: 13 out of 15 dogs had a significant regression of their tumors to less than 10% of the original volume; in 1 dog the tumor remained unchanged and in 1 dog it progressed. (3) In the third group, 8 dogs received 1 times 10-8 mitomycin- and VCN-treated tumor cells: the application of this cell dose resulted in an accelerated tumor progression in all 8 dogs, 3 of which have already died of metastasis. The significance of these findings, with respect to potentiation and abrogation of the immunological response and with regard to immunotherapy in man, is discussed.

Animals