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Biomedical subjects

M R Parwaresch

Publications and source records attributed to M R Parwaresch.

At least 109 records · Page 6Linked to original sources
PubMed1983

Combined immunohistochemical staining for surface IgD and T-lymphocyte subsets with monoclonal antibodies in human tonsils.

The aim of the present paper is to detect two different antigens simultaneously in a single slide. In cryostat sections of human tonsils, B-lymphocytes of follicle mantle-bearing surface IgD were immunostained with the alkaline phosphatase method using monoclonal anti IgD. The subsequent staining for T-lymphocyte subsets (T-helper and T-suppressor lymphocytes) was performed again with the alkaline phosphatase method using one of the monoclonal antibodies OKT 4, OKT 8, Leu 3a, Leu 2a. The best results with the alkaline phosphatase method were achieved using naphthol AS phosphate and Fast Blue BB for the revelation of the first antigen and naphthol AS-BI phosphate and diazotized New Fuchsin for the second.

Antibodies, Monoclonal↗
PubMed1983

Differential immunohistochemical resolution of the human mononuclear phagocyte system.

Four new monoclonal antibodies, termed Ki-M1, Ki-M2, Ki-M3, and Ki-M4, were developed for distinguishing macrophage subgroups. Purified lysosomes of cells of stimulated U-937 cell line were used as immunogen. Specificity control was performed by staining unfixed cryostat sections of fresh human tissue with an immunohistochemical method, which allowed reliable recognition of reactive structures. Ki-M1 reacted with macrophages of lymphoid tissue, lung, and serous cavities. Ki-M2 recognized Kupffer cells and splenic macrophages. Both monoclonal antibodies reacted with interdigitating reticulum cells and Langerhans cells, which are thought to be accessory cells of the T-cell immune response. Accessory cells of the B-cell immune response, on the other hand, showed reactivity with Ki-M3 and Ki-M4. Thus, analogous to T lymphocytes, the human mononuclear phagocyte system (MPS) can be subdivided into different subgroups with the aid of appropriate monoclonal antibodies.

Animals↗
PubMed1983

Monocytic origin of human alveolar macrophages.

The monocytic lysosomal acid esterase (AcE; EC 3.1.1.6) comprises five isoenzymes, each having specific isoelectric points (pI) as well as antigenicity. In the present study attempts were made to retrace the monocytic origin of human alveolar macrophages (AM) by comparison of their isoenzyme patterns with those of blood monocytes. Resident AM obtained from bronchial lavages lacking any neutrophils and unstimulated monocyte admixture showed in addition to the five monocytic isoenzymes nine additional isoenzyme loci. In vitro stimulation of blood monocytes (BM) using lymphokine-conditioned media led to a gradual transition of the typical monocytic isoenzyme pattern into that of AM. It is concluded that AM originates from blood monocytes by tissue-specific stimulation. This cellular transformation can be modeled in vitro as far as morphology, cytochemistry, and isoenzyme pattern are concerned.

Acetylesterase↗
PubMed1983

Monoclonal antibody Ki-M4 specifically recognizes human dendritic reticulum cells (follicular dendritic cells) and their possible precursor in blood.

Ki-M4, a new IgG3 monoclonal antibody, selectively recognizes dendritic reticulum cells (DRC; follicular dendritic cells) in all human lymphatic organs, as tested by the immunoperoxidase method on the light and electron microscopic level. This antibody was raised against separated lysosomes of the 12-0-tetradecanoyl phorbol-13-acetate (TPA) stimulated permanent cell line, U-937, derived from a human histiocytic lymphoma. No cross-reactivity was encountered in epithelial and mesenchymal cells, including macrophages and other cells detectable in bronchial and peritoneal lavages. In the nonadherent fraction of the mononuclear blood cells collected from the interphase of a Ficoll-Urografin gradient (density = 1.077 g/ml), 0.1 per million of the cells hitherto not classified as monocytes or lymphocytes showed a strong reaction. All other separated blood cell types were devoid of any reactivity. The observation that DRC share a highly restricted, and thus specific, antigen with a small but distinct subpopulation of mononuclear leukocytes implies their blood derivation.

Antibodies, Monoclonal↗
PubMed1983

Peroxidase-positive mononuclear leukocytes as possible precursors of human dendritic reticulum cells.

In this report, we provide evidence that suggests the dendritic reticulum cells (DRC) occurring in germinal centers of lymphatic follicles originate from a distinct endogenous peroxidase-positive mononuclear blood cell subset. The new monoclonal antibody Ki-M4 showed a highly restricted reactivity, tested by immune histochemistry, being confined to DRC, lining cells of lymph node sinuses, and to 0.001% of nonadherent mononuclear blood cells separated at a density of 1.077 g/ml; the latter exhibited endogenous peroxidase activity in cytoplasmic granules. Such granules being specific for myelomonocytic cells implies that DRC may derive from a committed nonadherent subpopulation of this cell line, which in turn originates from bone marrow. The Ki-M4-reactive antigen was found to be destroyed by the majority of fixatives and to resist 4% paraformaldehyde for 10 min and cold acetone for 30 sec. In cell suspensions from human tonsils, Ki-M4 showed strong reactivity with the outer surface of DRC plasma membrane. This observation demonstrates the possibility of using Ki-M4 to separate DRC from cell suspensions in functional tests.

Animals↗
PubMed1982

Analysis of acid esterase activity and polymorphism as an aid for the classification of human low-grade malignant non-Hodgkin's lymphomas.

Investigating a link between enzyme histochemical and recent immunohistochemical results, the authors studied the activity and the polymorphism of acid esterase (EC 3.1.1.6) in well defined human B-cell lymphomas. TWelve cases of chronic B-lymphocytic leukemia, 18 cases of centroblastic/centrocytic follicular lymphoma, and 17 cases of lymphoplasmacytic/lymphoplasmacytoid lymphoma, as diagnosed according to Kiel classification, were subjected to enzyme assay and isoelectric focusing of acid esterase. Enzyme values revealed no characteristic distribution among the lymphoma entities. The isoenzyme pattern, specific to normal human B-lymphocytes, were regularly detectable in all lymphoma entities. The results document the B-cell origin of the analyzed subsets of B-cell malignancies, although distinctive acid esterase patterns were lacking. The prevalence of three anodal isoenzymes in cases of chronic B-lymphocytic leukemia, hardly detectable in other lymphoma entities, were interpreted as the expression of a clonal proliferation, arrested at a certain differentiation stage of B-cells common for the majority of the tumor cells. Hence, further evidence is provided supporting the view that the studied entities represent B-cell neoplasias at different stages of differentiation expressing a variety of different markers.

B-Lymphocytes↗
PubMed1982

Enzymecytochemical heterogeneity of human chronic T-lymphocytic leukemia as demonstrated by reactivity to dipeptidylaminopeptidase IV (DAP IV; EC 3.4.14.4).

The observation that the dipeptidylaminopeptidase IV (DAP IV; EC 3.4.14.4) occurs exclusively in T mu lymphocytes, stimulated the following study on cases of chronic lymphocytic leukemias. Thirty cases of human chronic lymphocytic leukemia were subjected to the DAP IV reaction in addition to the usually applied cytochemical and immunological tests (acid alpha-naphthyl acetate esterase, acid phosphatase, rosetting with sheep erythrocytes and surface Ig). DAP IV activity was measured directly in normal granulocytes, monocytes, B and T lymphocytes as well as in lymphocyte suspensions of leukemic cases. On a cytochemical level granulocytes, monocytes, B lymphocytes and all 24 cases of B-CLL were found to be DAP IV negative, though some of the latter showed positive reactions for AcE and AcP. From the six cases of T-CLL, five were positive to DAP IV cytochemically. No discrepancies were observed between cytochemical and biochemical results. It is concluded that DAP IV is a reliable and easy to perform marker for T mu lymphocytes and their neoplasias. The results have been interpreted as further evidence for a clonal nature of T-cell neoplasias.

Acid Phosphatase↗
PubMed1982

The presence of isoenzymes in monocytes and macrophages.

The present paper gives a review of enzyme polymorphism found in individual human blood cells with special reference to human blood monocytes. Evidence is presented indicating that not only different blood cells but also their descendants are characterized by certain constellation of isoenzymes. Considering the typical polymorphism of acid phosphatase (EC 3.1.3.2) and acid esterase (EC 3.1.1.6), we could show that a reliable biochemical marker for blood monocytes can be worked out, which is also detectable in peritoneal and alveolar macrophages as well as epithelioid cells. In different examples we could demonstrate that the isoenzyme patterns of blood monocytes can be modified into that of peritoneal and alveolar macrophages by different stimulation procedures in vitro. Studying the enzyme variants in blood monocytes and different tissue macrophages, additional criteria have been collected, which document the monocytic origin and homogeneity of the human mononuclear-phagocytic system.

Acid Phosphatase↗
PubMed1982

Polymorphism of acid hydrolases in human epitheliod cells indicates cellular origin and defined differentiation stages.

The following work was planed to clarify whether enzyme variants were appropriate in disclosing cellular derivation and in indicating defined differentiation stages of stimulated cells. Light and electronmicroscopical studies have revealed a close relationship between monocytes, tissue macrophages, and epithelioid cells. In the present investigation direct evidence was provided for the monocytic origin of epithelioid cells. It could be shown that the monocyte specific isoelectric focusing patterns of acid phosphatase (EC 3.1.3.2) and nonspecific acid esterase (EC 3.1.1.6) were regularly shared by epithelioid cells, obtained from lymph nodes of patients with sarcoidosis. Concerning additional isoenzymes of epithelioid cells in comparison to blood monocytes and peritoneal macrophages, it is concluded that the analysis of enzyme polymorphism might be a reliable tool for tracing cell lineage and for defining functional stages during cell differentiation.

Acid Phosphatase↗
PubMed1982

Enzymehistochemical staining of T mu lymphocytes for glycyl-proline-4-methoxy-beta-naphthylamide-peptidase (DAP IV).

Human T mu lymphocytes are detected in a histochemical reaction with glycyl-proline-4-methoxy-beta-naphthylamide-peptidase (DAP IV; EC 3.4.14.4). DAP IV occurs in 59.4 +/- 8.25 of human T lymphocytes in blood and marrow. Direct biochemical assays were in accord with the cytochemical findings. DAP IV-positive lymphocytes accounted for 89.5 +/- 0.6% of T-lymphocytes bearing Fc receptor for IgG contained 11.3 +/- 8.3% T-lymphocytes bearing Fc receptor for IgM.

Cell Separation↗
PubMed1982

Isozymes of acid esterase and acid phosphatase in permanent human hematopoietic cell lines.

Net enzyme activities of normal human blood cells were measured, and isoelectric focusing patterns of acid esterase (AcE) (EC 3.1.1.6) and acid phosphatase (AcP) (EC 3.1.3.2) were compared with corresponding data obtained for two acute T-lymphoblastic leukemia cell lines (JM, Molt-4), one acute B-lymphoblastic leukemia cell line (Ball-1), one acute non-B-non-T-lymphoblastic leukemia cell line (KM 3), and one promyelocytic leukemia cell line (HL-60). The AcE isozymes, found in the individual blood cell types, were regularly expressed by the corresponding cell lines. AcP was regularly expressed by lines HL-60 and KM 3, but lines JM, Molt-4, and Ball-1 showed additional isozymes and/or reduction of the intensity of the typical isozymes found in the presumed normal counterparts. This phenomenon resulted partly in an obscuration of the typical isozyme patterns. Our study documents the applicability of isozyme mapping to the characterization of permanent hematopoietic cell lines. The results suggest that long-term culture conditions can repress phenotypic properties and/or derepress gene activities.

Acetylesterase↗
PubMed1981

Enzyme polymorphism in the classification of human malignant lymphoma.

Recent studies on the polymorphism of lysosomal hydrolases have shown that all individual blood cell types in the human being possess their own isoenzyme pattern. In the present study acid phosphatase activity of normal human B-lymphocytes and of four different types of low-grade malignant non-Hodgkin's lymphomas according to the Kiel classification was estimated. In addition, the isoenzyme pattern of AcP was investigated by isoelectric focusing. Chronic lymphocytic leukemia of B-cell type (N = 9) and centroblastic centrocytic follicular lymphoma (N = 10) demonstrated significantly lower values than lymphoplasmacytic/lymphoplasmacytoid lymphomas (N = 28) and plasmacytomas (N = 8). The isoenzyme pattern of normal human B-lymphocytes comprised 12 bands between pH 6.3 and 3.85. This basical pattern was shared by all four lymphoma entities. Only lymphoplasmatic lymphoplasmacytoid lymphoma and plasmacytoma revealed additional bands, which probably account for the higher net enzyme activity in these cases.

Acid Phosphatase↗
PubMed1981

The homogeneity and monocytic origin of human peritoneal macrophages evidence by comparison of esterase polymorphism.

The aim of this study was to test the hypothesis of the homogeneity and monocytic origin of the normal unstimulated (resident) peritoneal macrophages in man. The isoelectric focusing pattern of one of the lysosomal marker enzymes, acid esterase (EC 3.1.1.6), was studied in highly purified samples of normal human blood cell fractions. All types of blood cells showed a cell-specific constellation of their enzyme variants as far as blood cells were concerned. Comparisons were made with resident peritoneal macrophages of healthy subjects. Only blood monocytes shared all their isoenzymes with peritoneal macrophages. Two additional bands, not seen in monocytes, occurred, however, in peritoneal macrophages. During exposure of blood monocytes to prolonged glass adherence there was a gradual transition of the monocytic pattern into that of peritoneal macrophages. The results document the homogeneity and monocytic origin of resident peritoneal macrophages in man.

Ascitic Fluid↗