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J Gerdes

Publications and source records attributed to J Gerdes.

At least 199 records · Page 11Linked to original sources
PubMed1982

Ultrastructural localization of human complement C3b receptors in the human kidney as determined by immunoperoxidase staining with the monoclonal antibody C3RTo5.

The precise localization of human complement C3b receptors in human kidneys was investigated by means of indirect immunoperoxidase staining of frozen kidney sections with the monoclonal antibody C3RT05, which is specific for human C3b receptors. Transmission electron microscopy indicated that C3b receptors are located on the entire cell membrane of the visceral epithelial cells of the glomeruli.

Antibodies, Monoclonal↗
PubMed1982

The value of immunohistological screening in the production of monoclonal antibodies.

This paper describes an immunoperoxidase technique for labelling cryostat tissue sections which is routinely used in the authors' laboratories both in the initial screening of hybridoma culture supernatants, and also during the subsequent cloning and growth of antibody-secreting cell lines. The technique can readily be performed on 100 samples in less than 3 h and is free of non-specific background labelling. The staining pattern of a monoclonal antibody on a single tissue section allows semiquantitative assessment of its reactivity against a wide variety of tissue constituents and is thus inherently much more informative than conventional screening techniques (such as binding assays) which yield only a single numerical value for each test performed. In consequence it is often possible to identify the probable specificity of a new monoclonal antibody at the primary screening stage. A further important advantage of immunohistological screening is that it detects antigens on cells or other tissue structures which do not readily enter suspension and also antibodies against nuclear and cytoplasmic antigens. Examples of monoclonal antibodies analysed by immunohistological screening include antibodies against C3b receptor, HLA-DR, factor VIII-related antigen, human syncytiotrophoblast, dendritic reticulum cells and a proliferation-associated cell surface glycoprotein.

Animals↗
PubMed1982

Characteristics of Hodgkin's disease-derived cell lines.

In the last 3 years we were able to establish five long-term in vitro cell cultures from biopsy specimens taken preterminally from four patients with histologically proven Hodgkin's disease (nodular sclerosing type, clinical stage IVB). Four of the lines are continuously proliferating in vitro; one culture stopped growth for unknown reasons after 7 months. When culture conditions were modulated, the first culture, L 428, gave rise to two sublines: L 428 KS, after adaptation to calf serum, and L 428 KSA, permanently growing as an adherent monolayer line after treatment with a phorbol ester (12-O-tetradecanoylphorbol-13-acetate) for 3 weeks. Cell-marker analysis by conventional means (SIg, cIg, rosette formation, Epstein-barr virus reactivity, cytochemistry, phagocytosis, and lysozyme production) and with monoclonal antibodies directed against various human lymphoid, myeloid, and monocytoid antigens showed that the tested cell lines are clearly different from all hitherto described hematopoietic lines; they most likely represent a cell type resembling an early myeloid-monocytoid progenitor cell. Conditioned medium of the L 428 cells and its two sublines showed colony-stimulating factor activity and suppression of spontaneous cell-mediated cytolysis of L 428 KS and K 562 cells.

Adult↗
PubMed1982

Complement (C3) receptors on dendritic reticulum cells of normal and malignant lymphoid tissue.

The aim of this study was to investigate whether dendritic reticulum cells (DRC) in normal lymphoid tissue and in malignant non-Hodgkin's lymphomas possess receptors for the third complement component (C3 receptors). For this purpose we studied the in situ expression of C3 receptors in lymphoid tissue by staining frozen tissue sections with an antiserum specific for human C3 receptors (AC3RS), using a modified immunoperoxidase method. The results indicate that DRC of normal and malignant lymphoid tissue express large amounts of C3 receptors. Absorption experiments revealed that the C3 receptors of DRC are identical with, or at least share common antigenic sites with, the C3 receptors expressed by tonsil B cells.

Child↗
PubMed1982

Human complement (C3b) receptors defined by a mouse monoclonal antibody.

The aim of the present study was to prepare a monoclonal antibody against human C3 receptors. The monoclonal antibody termed C3RTo5, which is characterized in this study, inhibited the ligand binding of C3b receptors of human erythrocytes, neutrophils and lymphocytes, but did not block the ligand binding of C3bi and C3d receptors. C3RTo5 reacted only with cells that express C3b receptors. It did not react with cells that do not express C3 receptors or with cells that express C3 receptors other than C3b receptors. The reactivity of C3RT05 against C3b receptors of human erythrocytes, neutrophils, glomerular cells, tonsil cells, or spleen cells could be removed by absorption with human erythrocytes or tonsil cells, whereas absorption with human peripheral T cells or sheep erythrocytes had no effect. In immunoprecipitation studies, a glycoprotein with a mol. wt of 205,000 could be isolated with C3RTo5 from non-ionic detergent lysate of tritiated tonsil cells. A rabbit antiserum prepared against this glycoprotein was able to stain C3b receptor-positive cells, inhibit C3b receptor ligand-binding activity and, furthermore, to precipitate a 205,000 mol. wt component. The results of this study indicate that C3RTo5 is a monoclonal antibody with selective reactivity to C3b receptors and presumably to the binding sites within the receptor molecule. Using C3RTo5 further strong evidence was obtained that membrane-bound C3b receptors have a mol. wt of 205,000.

Animals↗
PubMed1981

Hodgkin and sternberg-reed cell antigen(s) detected by an antiserum to a cell line (L428) derived from Hodgkin's disease.

Antisera to the cell line L428, derived from Hodgkin's disease, were raised in rabbits by injecting L428 cells intravenously and subcutaneously. The anti-L428 cell serum that did not react with HLA-DR was absorbed with tonsil cell plus acute myeloid leukemia cells or tonsil cells plus neutrophils, monocytes, and blood lymphocytes. Then it was tested for its ability to discriminate between L428 cells, Hodgkin and Sternberg-Reed cells, and various other cells. It was found that the anti L428 cell serum absorbed with tonsil cells plus acute myeloid leukemia cells stained only L428 cells, Hodgkin and Sternberg-Reed cells, and neutrophils. The anti L428 cell serum absorbed with tonsil cell plus neutrophils, monocytes, and blood lymphocytes reacted with L428 cells and Hodgkin and sternberg-Reed cells from 13 cases of Hodgkin's disease. It did not react with any other cell type present in the blood or in lymphoid tissue or with cells from five cases of non-Hodgkin's lymphoma. The absorbed anti-L428 cell serum also failed to stain Daudi and HRIK cell line cells. We conclude that the anti-L428 cell serum defines an antigen that is apparently restricted in expression to L428 cells and Hodgkin and Sternberg-Reed cells. This is a strong indication that the L428 cell line cells are derived from Hodgkin and Sternberg-Reed cells.

Antigens, Neoplasm↗
PubMed1981

Immunohistological demonstration and physiochemical characterization of C3 receptors of normal and neoplastic lymphoid cells.

The in situ distribution of C3 receptor-positive cells was analyzed by immunoperoxidase staining of normal and malignant lymphoid tissue with an anti-C3 receptor serum (AC3RS). AC3RS immunostaining coincided well with the pattern found previously with the erythrocyte-antibody-complement complex (EAC) adherence technique. This indicates that AC3RS is a new tool for the detection of C3 receptors in situ. In addition, evidence is presented that the different results obtained in molecular weight analyses of human C3 receptor molecules may be due to different preparation procedures. When human tonsillar lymphocyte membrane fractions were solubilized, and then radioiodinated, subunits with apparent molecular weights of 38,000 and 18,000 could be precipitated from these lysates with AC3RS. When whole cells were tritiated and then solubilized, only one component with an apparent molecular weight of 200,000 could be isolated with AC3RS.

Autoradiography↗
PubMed1981

Molecular weight analysis of Fc gamma-binding proteins of lymphoid leukemia, myeloid leukemia, and hairy-cell leukemia.

The molecular weights of EDTA-mercaptoethanol-soluble Fc gamma-affined proteins isolated from chronic lymphocytic leukemia of the B type, prolymphocytic leukemia of the B type, chronic myeloid leukemia and hairy-cell leukemia were compared. SDS polyacrylamide gel electrophoresis of the Fc gamma-binding material obtained from all six cases of B type leukemia revealed a single peak with an apparent molecular weight of 28,000. The Fc gamma-affined material isolated from the cells of two cases of chronic myeloid leukemia showed two peaks, one with an apparent molecular weight of 42,600 and one with an apparent molecular weight of 18,800. The Fc gamma-affined material isolated from the cells of two cases of hairy-cell leukemia electrophoresed in the form of a closely spaced double peak. One component of the double peak had an apparent molecular weight of 28,000 and thus corresponds to the Fc gamma-binding material of leukemic B cells. The second component had a slightly lower molecular weight. The latter component is not present on either leukemic B cells or myeloid cells. The results indicate that the EDTA-mercaptoethanol-soluble Fc gamma-affined proteins of different types of cells differ in molecular weight, and thus in molecular structure.

B-Lymphocytes↗
PubMed1981

Hodgkin's disease: establishment and characterization of four in vitro cell lies.

Four in vitro cell lines (L 428, L 439, L 538, and L 540) were established from different materials of three patients with Hodgkin's disease: pleural effusions, peripheral blood, and bone marrow. The histological diagnosis was confirmed in all cases by several independent histologists. All four cell lines have been in culture for over 6 months up to over 3 years. The neoplastic nature of the culture cells is indicated by the demonstration of several structural and numeric chromosome abnormalities associated with a monoclonal pattern of marker chromosomes. EBV-specific antigens (EBNA, VCA) were not detected in either cell line. Ia-like antigens, receptors for human T cells, acid phosphatase, and acid esterase were showen to be present in the cultured cells. All cell lines lacked surface or cytoplasmic Ig, HTLA, receptors for C3b, C3d, IgG-Fc, mouse E or sheep E, and were devoid of lysozyme, peroxidase, and chloracetate esterase. The described features do not represent B cells, T cells, myeloid cells, monocytes, or macrophages. The morphology and the marker pattern of the culture cells, however, is identical with that of freshly obtained Hodgkin's (H)- and Sternberg-Reed (SR)-cells, except for the lack of CIg in the in vitro cells, which is explained by the culture conditions. Heterotransplantation in nude mice was achieved by intracranial inoculation and by s.c. transplantation of cultured cells embedded in a plasma clot. The described findings suggest that these cultured Hodgkin's cell lines are indeed derived from H and SR cells. The cellular origin of these cells is not clear, the loss of cellular differential markers during the process of possible dedifferentiation is discussed.

Adult↗
PubMed1981

Immunohistological analysis of Hodgkin's and Sternberg-reed cells: detection of a new antigen and evidence for selective IgG uptake in the absence of B cell, T cell and histiocytic markers.

To help clarify the origin and nature of Hodgkin's (H) and Sternberg-Reed (SR) cells, three different sets of experiments were performed. First, it was shown that cytoplasmic gamma, kappa, lambda occur not only in H and SR cells, but also in polymorphic tumor cells of epithelial, neurogenic, and lymphoid origin. Furthermore, human IgG that was injected i.v. into rats penetrated many rat liver cells, whereas injected human alpha 1-antitrypsin did not. Second, staining of frozen sections revealed that H and SR cells lack surface immunoglobulin and peripheral T-cell antigen. Third, an antiserum raised against the L 428 cell line (derived from Hodgkin's disease) and absorbed with human serum and normal cells did not react with any cells of tonsil tissue (lymphoid cells, histiocytes, and interdigitating reticulum cells), whereas it reacted strongly with the L 428 cell line cells and with H and SR cells of 10 different cases. In all ten cases, the antiserum stained the surface of H and SR cells; in two cases, it also stained the nucleoli and some chromatin spots in H and SR cells. The results obtained in these experiments indicate that H and SR cells are not closely related to lymphoid cells, histiocytes, or interdigitating reticulum cells. The findings also suggest that H and SR cells express one or more antigens that have not yet been detected on or in normal cells.

Animals↗
PubMed1981

Human membrane-bound C3 receptors. I. Serological and immunohistological demonstration of C3 receptors.

The aim of the present study was to present further evidence of the specific reactivity of an anti-C3 receptor serum (AC3RS), to demonstrate membrane-bound C3 receptors by using this AC3RS in different serological an immunohistological methods, and to investigate the relationship between membrane-bound C3 receptors and alpha1-antitrypsin. The AC3RS, or F(ab')2 fragments of the IgG fraction of this antiserum, stained a percentage of various viable cell populations roughly equivalent to the number of cells that bound EAC3b and/or EAC3d; C3 receptor-negative T cells and thymocytes were not stained. On frozen sections of tonsils and kidneys it was found that the AC3RS stained area to which EAC3b adhered. After absorption with neutrophils or Ehu, the AC3RS inhibited the agglutination of EAC3d with tonsil cells, but not the agglutination of tonsil cells or neutrophils with EAC3b; this absorbed AC3RS still stained tonsil cells but not neutrophils, and in frozen tonsil sections it stained only those areas to which EAC3d adhered. The absorbed AC3RS did not stain glomeruli. Antisera to alpha1-antitrypsin failed to inhibit EAC agglutination with C3 receptor-bearing cells or to stain C3 receptor-positive cells either in suspension or in frozen sections. Absorption of th AC3RS with purified alpha1-antitrypsin did not affect its specific reactivity.

Antigen-Antibody Complex↗
PubMed1981

Human membrane-bound C3 receptors. II. Physiochemical properties of the C3b and C3d receptors isolated from tonsil cells by immunoprecipitation.

The aim of the present study was to define the physicochemical structure of C3b and C3d receptors of lymphoid cells. C3b and C3d receptors were isolated from KBr lysates of the 20,000 g fraction of human tonsil homogenates by immunoprecipitation with an anti-C3 receptor serum (AC3RS). Sodium dodecyl sulphate (SDS) gel filtration and polyacrylamide gel electrophoresis (PAGE) of unreduced immunoprecipitates revealed a highly predominant component with an apparent molecular weight (mol. wt.) greater than 1 x 10(6) and a small component with a mol. wt. of 80,000. After reduction, the SDS-PAGE profile was made up of a constant major 38,000 mol. wt. component and a inconstant smaller 18,000 mol. wt. component. The 38,000 (and also the 18,000) component could be isolated only from C3 receptor-active lysates, and not from C3 receptor-negative lysates. Taken together, the results of this study suggest that the active C3 receptor molecule of tonsil cells is a lipoprotein complex with a mol. wt. greater than 1 x 10(6); its protein moiety consists predominantly of disulphide-bridged polypeptide chains with a mol. wt. of 38,000; C3b and C3d receptors are composed of equal-sized polypeptide chains, but the specific binding sites for C3b and C3d are located on different molecules.

Chromatography, Gel↗
PubMed1980

Immunohistologic analysis of the organization of normal lymphoid tissue and non-Hodgkin's lymphomas.

Hoping to improve the systems for identifying and classifying normal and malignant lymphoid subpopulations, frozen and paraffin sections of nonmalignant lymphoid tissue and of malignant lymphomas were immunostained for surface (S) and cytoplasmic antigens using the peroxidase-antiperoxidase method. Primary follicle cells and follicle mantle cells known to be part of the recirculating B-cell pool were found to be constantly Ia and C3 receptor (C3R) positive, mostly SIgM and SIgD positive and cytoplasmic immunoglobulin (CIg) negative. The light zone of germinal centers (GC), which is rich in centrocytes, contained a large number of T cells and showed the well-known intercellular Ig network pattern; the dark zone, containing densely packed centroblasts, was usually free of T cells, but was bordered ay a mantle-like accumulation of T cells. Usually only some of the GC cells were definitely positive for SIg and CIg of different classes. All cells reacted positively for Ia and C3R. In areas described by other authors as containing marginal zone cells, cells densely bearing SIgM and deficient in SIgD were detected. The immunoblasts of the hyperplastic plasma cell reaction usually contained CIg. Cells from chronic lymphoid leukemia sections that immunostained for SIgM and SIgD were interpreted as representing a neoplasm of recirculating B cells expressing SIgM and SIgD. The immunohistologic architecture of follicular centroblastic/centrocytic lymphoma showed a more or less close similarity to the organization of secondary follicles. Lymphomas whose cells resembled reactive centrocytes were strongly SIgM positive and SIgD negative or only weakly SIgD positive. CIg was demonstrable in nearly 90% of the lymphomas whose cells resembled centroblasts and in 70% of the lymphomas whose cells resembled immunoblasts of the plasma cell reaction. Finally, immunohistologic staining results from a T-zone lymphoma are presented, which confirm that this lymphoma was composed of a neoplastic T zone and a non-malignant B zone.

Antigens↗
PubMed1980

Xenoantiserum to human C3 receptors: its preparation and effect on the C3b and C3d receptors of tonsil cells and the C3b receptors of erythrocytes and neutrophils.

Antisera directed against complement (C3) receptors on human tonsil cells were prepared and tested for their capacity to block specifically C3 receptors on various types of human cells. The antisera were capable of blocking both membrane-bound and solubilized C3 receptors of human tonsil cells. The C3b receptors of human erythrocytes and granulocytes were also blocked by the anti-C3 receptor sera. Sheep erythrocyte rosette formation was not affected. IgG-EoxA rosette formation was only slightly reduced by the anti-C3 receptor sera. Immunofluorescent staining with anti-C3 receptor sera revealed only a faint or negative staining of T cells and a distinct staining of EAC-reactive tonsil cells, lymphocytic leukaemia cells, and granulocytes. Absorption of the antisera with human serum proteins, brain, thymus, liver, EU-1 cell line cells, or trypsinized tonsil cells did not influence the capacity of the anti-C3 receptor sera to inhibit C3 receptors, whereas absorption with splenic tissue or tonsil cells completely removed the blocking activity of the anti-C3 receptor sera. Absorption with human erythrocytes or kidney removed only the inhibitory effect of the antisera on C3b receptors of tonsil cells, human erythrocytes, and granulocytes, but not on C3d receptors of tonsil cells. The results indicate that (a) the antisera prepared with the described procedure contained significant amounts of antibody against C3 receptors, (b) the receptors for C3b and C3d differe in antigenicity, and (c) the C3b receptors of tonsil cells, human erythrocytes, granulocytes, and probably glomerular cells have common antigenic sites.

Complement C3↗
PubMed1980

Physicochemical characterization of C3b receptors isolated from human erythrocytes by immunoprecipitation.

A high yield of active C3b receptors was obtained by solubilizing human erythrocyte membranes with 2 M KBr, whereas other solubilization agents yielded no, or significantly less activity. Gel filtration of the KBr lysates revealed that the apparent molecular wieght of biologically active C3b receptor molecules was greater than 1 x 10(6). Immunoprecipitates prepared with radio-iodinated KBr lysates and anti-C3 receptor sera (AC3RS) were subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) or sodium dodecyl gel filtration. Unreduced SDS-PAGE and gel filtration profiles showed three predominant peaks with apparent mol. wts of 1--1.3 x 10(6), 80,000 and 60,000. Whereas the high mol. wt component decreased only slightly after reduction, the 80,000 and 60,000 mol. wt components disappeared and two new peaks with apparent mol. wts of 38,000 and 18,000 appeared in SDS-PAGE profiles. Although the high mol. wt component present in reduced SDS-PAGE profiles was detectable in some of the control experiments, none of the other peaks could be precipitated with control sera, and these components could be demonstrated only when KBr lysates of C3b receptor-positive erythrocytes and AC3RS that were able to inhibit ligand binding of the C3b receptors were used for precipitation. These findings suggest that (a) the C3b receptor of human erythrocytes in its biologically active state is a macromolecule with an apparent mol. wt higher than 1 x 10(6) and (b) the protein moiety consists predominantly of non-covalently linked protein molecules with apparent mol wts of 80,000 and 60,000. These protein molecules are composed of disulphide-bridged polypeptide chains with apparent mol. wts of 38,000 and 18,000.

Chromatography, Gel↗