Biomedical subjects

M R Parwaresch

Publications and source records attributed to M R Parwaresch.

At least 91 records · Page 5Linked to original sources

PubMed1986

Induction of hypoxanthine phosphoribosyltransferase deficiency in human U-937 cells.

The aim of the study was to establish enzyme-deficient mutants of the human permanent cell line U-937. Following chemical mutagenesis with the use of ethyl methanesulfonate, this cell line was chronically exposed to increasing concentrations of the toxic hypoxanthine analogue 6-thioguanine. Cells surviving hypoxanthine-aminopterin-thymidine selective media were separated by glass adherence with the use of 12-O-tetradecanoyl-phorbol-13-acetate. Three mutant clones were established, which have remained hypoxanthine phosphoribosyltransferase (HPRT) deficient for a period of 7 months, as shown by indirect measurements with the use of autoradiography and scintillation counting of cells exposed to [3H]hypoxanthine. Since the phenotypic properties and growth behavior of U-937 cells have remained unaltered after the induced mutation, a highly restricted chromosomal segment coding for HPRT seems to have been mutated.

Autoradiography↗

PubMed1986

Monocyte/macrophage-reactive monoclonal antibody Ki-M6 recognizes an intracytoplasmic antigen.

A monoclonal antibody, termed Ki-M6, is described, which shows a restricted reactivity to cells of the monocyte/macrophage system. On light- and electron-microscopic immunoperoxidase staining Ki-M6 recognizes monocytes and the phagocytosing compartment of macrophages residing in different tissue sites; granulocytes and the so-called immune accessories of B- and T-cell immune response as closely monocyte/macrophage related cell populations do not reveal any reactivity. This is shown by comparison with the monoclonal antibodies Ki-M4 and Ki-M1 or OKT6 recognizing immune accessory cells by immunohistochemical methods. Ki-M6 binds to a lysosomal membrane-restricted antigen of 60,000 daltons without influencing significantly lysosome-related functions as far as the chemiluminescence response is concerned.

Antibodies, Monoclonal↗

PubMed1985

Establishment of thymidine kinase (EC 2.7.1.21)-deficient mutants of human monocytic cell line U-937.

Using morphologic, enzyme-cytochemical, and immunocytochemical methods, the functional diversity of the mononuclear phagocyte system can be studied only to a limited extent. Therefore, enzyme-deficient monocyte/macrophage cell lines have been established as technical prerequisites for the generation of monocyte/macrophage hybrids by applying selective media. After mutation with ethylmethanesulfonate, six clones of U-937 were selected against increasing concentrations of 5'-bromodesoxyuridine; these clones are defective in thymidine kinase (EC 2.7.1.21), as shown by autoradiography and direct measurement of [3H]thymidine uptake. A broad marker panel indicates that the clones could be appropriate for the establishment of human monocyte/macrophage hybrids.

Cell Line↗

PubMed1985

Ki-M2R, a new specific monoclonal antibody, discriminates tissue macrophages from reticulum cells and monocytes in vivo and in vitro.

Utilizing rat peritoneal macrophages as the immunogen, a new monoclonal antibody enabling differential monitoring of the mononuclear phagocyte system (MPS) by immunohistochemistry has been raised. Designated Ki-M2R, this antigen could be detected with the immune alkaline phosphatase reaction on all macrophages including those of bone marrow, lymphatic sinuses, lymphoid follicles, splenic red pulp, and von Kupffer cells of the liver, as well as on macrophages of connective tissue, renal interstitial tissue, serous cavities, and gastrointestinal tract. Langerhans cells--the MPS-derived reticulum cells of the epidermis--interdigitating reticulum cells, and dendritic reticulum cells of lymphoid follicles were invariably negative. Blood monocytes were rendered positive only after evolving into macrophages upon appropriate stimulation. Thus, Ki-M2R selectively labels monocytes after transformation into macrophages.

Animals↗

PubMed1985

Bone marrow findings in systemic mastocytosis.

The neoplastic proliferation of tissue mast cells constitutes a group of rare diseases that have localized and systemic variants. The cytologic (n = 7) and histologic (n = 38) findings in bone marrow from a total of 45 patients with systemic mastocytosis were evaluated. Three distinct histologic patterns of marrow involvement were distinguished. In 21 cases a patchy or focal infiltration pattern was encountered. Mast cell aggregates were located predominantly in peritrabecular and perivascular areas. The adjacent trabeculae were thickened. A dense network of reticulin fibers and foci of lymphocytes accompanied the mast cell infiltrates. Increased numbers of eosinophils frequently demarcated the mast cell infiltrates from the surrounding tissue. In the noninfiltrated marrow areas hematopoiesis and the distribution of fat cells appeared to be normal. This histologic pattern, designated type 1, was observed exclusively in patients showing primary involvement of the skin, indistinguishable from urticaria pigmentosa. In 14 additional cases peritrabecular and perivascular sheets of mast cells, with concomitant fibrosis and osteosclerosis, were also present. Unlike the findings in type 1, however, the noninfiltrated marrow areas showed marked reductions in fat cell content and markedly increased granulocytopoiesis or increased numbers of blast cells (infiltration pattern type 2). On the basis of the hematologic and clinical findings, chronic myeloid leukemia was diagnosed in six of these cases, myelomonocytic leukemia in three cases, and acute myeloid leukemia in two cases. The bone marrow of three patients was diffusely infiltrated by atypical mast cells, leading to marked hypoplasia of fat cells and blood cell precursors. These histologic features were identified as infiltration pattern type 3. The diagnosis of mast cell leukemia was confirmed in all three cases by the presence of numerous mast cells in the blood. The prognosis for patients with the type 1 marrow infiltration pattern and primary skin involvement was favorable (actuarial survival rate five years after diagnosis, 0.75). This variant was called benign systemic mastocytosis. Primary skin involvement did not occur in the patients with type 2 or 3 infiltration patterns. The prognosis for these patients was poor (actuarial survival five years after diagnosis, 0.17 for type 2 and 0.00 for type 3). These two forms were accordingly designated malignant systemic mastocytosis.

Adult↗

PubMed1985

Tissue mast cells in health and disease.

Mast cells (MC) are regular constituents of soft tissue and occur with varying frequency in nearly every organ. They derive from monocytic cells occurring in the adhering mononuclear fraction of the peripheral blood. Their subsequent evolution into mature MCs is primed by a MC generating lymphokine released by sensitized T-cells on restimulation by the antigen. MC granules contain preformed heparin histamine and eosinophil chemotactic factors. Other factors such as leukotriene B4 can be produced by MCs following stimulation. This is the case during the initial phase of nonspecific inflammations, when MCs are stimulated by complement activation. In the immediate type hypersensitivity reaction giving rise to IgE, MC degranulation occurs independent from complement. In IgG and IgM mediated reactions, however, MC involvement is effected by complement consumption and C5 a generation. In delayed type hypersensitivity MCs increase locally. Their functional significance remains obscure. MC neoplasias are rare and generally confined to the dermis. Cutaneous mastocytoses are called benign mastocytoma when localized and urticaria pigmentosa when disseminated. Generalized mastocytosis involves extracutaneous tissue irrespective of skin involvement. Those associated with urticaria pigmentosa-like skin lesions, present at the onset of the disease, have a significantly higher survival rate than those lacking a primary skin involvement. The term urticaria pigmentosa should be reserved for cases of cutaneous mastocytosis without extracutaneous involvement. Cases of mastocytoses lacking primary skin lesions assume a malignant course and are additionally aggravated by high incidence of myeloproliferative disorders and MC leukemia. MC sarcoma is an extremely rare neoplasia of MCs which may also terminate as a MC leukemia.

Bone Marrow↗

PubMed1985

Bimodal differentiation prospectives for promyelocytes.

In accordance with the long-range goal to demonstrate the direct derivation of blood monocytes from promyelocytes in human bone marrow, the promyelocytic cell line HL-60 in the present study was differently stimulated with various inducers for the purpose of documenting its capability to evolve into granulocytes or monocytes-macrophages. Each differentiation line was monitored with the use of marker enzymes, antigens, and cell-specific monoclonal antibodies. In addition, studies were done on normal human bone marrow and leukemias. The results provided strong evidence for the close cytogeneic relationship of granulocytes and monocytes and for a common progenitor at the maturation stage of promyelocytes-myelocytes for both cell lineages.

Antibodies, Monoclonal↗

PubMed1984

Blood findings in lymphogranulomatosis X.

A quantitative analysis of leukocytes in the peripheral blood of 16 patients with lymphogranulomatosis X revealed the following findings: The number of total leukocytes, monocytes, neutrophils, and eosinophils varied considerably and showed no clear common tendency. Blood basophils were found to be clearly increased with a mean value of 0.17 +/- 0.18 X 10(9)/1 as compared to 55 normal donors (0.04 +/- 0.01 X 10(9)/1). Lymphocyte counts were within the normal range. Lymphopenia was seen in 2 patients. Lymphocytes with azure granules which encompass T-suppressors were found to exceed normal values by a factor of 15. In 14 cases a variable number of hyperbasophil cells were present. Lymphogranulomatosis X seems to present with a characteristic blood picture which might be of diagnostic aid.

Adult↗

PubMed1984

Immunoelectron microscopic demonstration of tissue antigens with monoclonal antibodies.

Two methods of demonstrating tissue antigens by ultrastructural enzyme immunohistochemistry were tested. The monoclonal antibodies Ki-M1 and Ki-M4 were chosen for testing the methods because Ki-M1 identifies a relatively stable, and Ki-M4 a very unstable antigen. The two antibodies react selectively with human macrophages and interdigitating reticulum cells or dendritic reticulum cells of lymphoid follicles. The Ki-M1 reaction product is confined to the surface membrane. Ki-M4 reactivity is located on the surface membrane and, less often and to a lesser extent, in the cytoplasm. The technical prerequisites for reliable conservation of the antigens identified by these two antibodies were standardized. The results indicated that prior fixation in 4% paraformaldehyde is preferable for optimum preservation of stable antigens. Application of the primary antibody prior to fixation was found to be the best procedure for demonstrating unstable antigens, although nonspecific reactions were seen more often with this method.

Antibodies, Monoclonal↗

PubMed1984

Alternative myelomonocytic differentiation of HL-60 reflects dual prospective potency of promyelocytes in human.

The permanent promyelocytic cell line HL-60 was subjected to stimulation with dimethyl sulfoxide (DMSO) and retinoic acid (RA), as well as 12-O-tetradecanoylphorbol-13-acetate (TPA) and lymphokine conditioned media for the induction of granulocytic or monocytic differentiation, respectively. Cells were investigated cytochemically using alpha-naphthylacetate esterase (acid esterase; AcE), naphthol AS-D chloroacetate esterase, and peroxidase reactions. In addition, the granulocyte or monocyte specific isoenzyme patterns of AcE as an intracytoplasmic property and the immunoreactivity to monoclonal antibodies recognizing granulocytes and monocytes (Ki-M2, Ki-M5) or monocytes alone (Ki-M1) were considered. The results indicated that HL-60 cell line bear the potency to evolve into granulocytes as well as monocytes. Additional studies performed on normal human bone marrow stained for AcE led to the conclusion that the myeloid cell line remains bipolar until the maturation stage of promyelocytes. Myelocytes being AcE positive only in 11.5 +/- 5.0 are heterogeneous and display the first indications of separated monocytic or granulocytic differentiation.

Antibodies, Monoclonal↗

PubMed1984

Cytochemical distribution of dipeptidylaminopeptidase IV (DAP IV; EC-3.4.14.5) in T-lymphoblastic lymphoma/leukemia characterized with monoclonal antibodies.

In human blood and bone marrow, dipeptidylaminopeptidase IV (DAP IV; EC 3.4.14.5) selectively occurs in T lymphocytes bearing Fc receptors for IgM. In the present study 35 cases of lymphoblastic lymphoma and leukemia were analysed for the specificity, incidence and reaction pattern of DAP IV. On the basis of immunohistochemical staining with monoclonal antibodies and enzyme cytochemical staining for acid phosphatase, 12 cases were classified as B-type neoplasms. In 23 cases T-cell properties were expressed to different extents, apparently reflecting different categories of maturation. Whereas B-cell lymphomas were invariably negative for DAP IV, seven of the 23 T-lymphoblastic lymphomas/leukemias showed this enzyme. Thus DAP IV is a highly specific marker for a distinct T-cell subpopulation, apparently irrespective of the stage of differentiation.

Acid Phosphatase↗

PubMed1984

Origin and kinetics of resident tissue macrophages. Parabiosis studies with radiolabelled leucocytes.

To elucidate the origin and renewal kinetics of peritoneal macrophages, as a typical example of the mononuclear phagocytic system, syngeneic rats were treated with tritiated thymidine [3H]TdR and leucocytes were transferred to unlabelled recipients over a bilateral arteriovenous shunt. Labelled and unlabelled monocytes were evenly distributed in both animals as shown by autoradiography. It was ascertained that no 'autoradiographically' detectable reutilization of label occurred and that transferred cells showed undisturbed kinetics. The results imply: resident peritoneal macrophages derive from blood monocytes; peritoneal macrophages represent a homogeneous population in respect to their cellular origin; blood monocytes as a myelogenous cell line do not represent a generative end cell. They migrate into the tissue (peritoneal cavity) and differentiate into resident macrophages, undergoing on average one mitosis per cell during a period of approximately 7 days. resident peritoneal macrophages are derived 50% from blood monocytes and 50% from division in situ; and under steady-state conditions the renewal rate amounts to 0.18%/h, which yields a half-life time of 16 days and a renewal time of 23 days.

Animals↗

PubMed1984

Monocyte/macrophage-specific monoclonal antibody Ki-M1 recognizes interdigitating reticulum cells.

A monoclonal antibody, Ki-M1, was produced, and its immunoreactivity was tested by light and electron-microscopic immunohistochemistry. Ki-M1 was found to react with monocytes, cells of the mononuclear phagocyte system (MPS), interdigitating reticulum cells (IDC), and the so-called indeterminate dendritic cells of lymphoid tissue. No reactivity was seen in other human tissues or other hematopoietic cells, including granulocytes and cells of the unstimulated promyelocyte cell line HL-60. Thus, Ki-M1 is the first of the monoclonal antibodies to MPS cells to react with both human IDC and MPS cells. This suggests that IDC and MPS cells may have a common cytogenesis.

Animals↗

PubMed1983

Subtyping of chronic lymphocytic leukemia of T-type by dipeptidylaminopeptidase IV (DAP IV), monoclonal antibodies, and Fc-receptors.

The authors have recently provided evidence for a restricted occurrence of the enzyme dipeptidylaminopeptidase IV (DAP IV) in normal human Tmu lymphocytes as far as blood and bone marrow cells are concerned. In this report, this issue has been extended to cases of chronic lymphocytic leukemia (CLL) along with T-monoclonal antibodies of OKT series (OKT4, OKT8) and Fc receptors for IgM (Tmu) and IgG (Tgamma). Seven of eight cases of T-CLL were DAP IV-positive, whereas all B-CLL cases like normal B-cells were invariably enzyme-negative. Parallel studies with OKT antibodies and Fc receptors showed that the cases positive to DAP IV also revealed OKT4 reactivity and, to some extent, Fc IgM receptor. In a single DAP IV-negative T-CLL with azurophilic granules, the leukemic cells were shown to be reactive for OKT8 and bore Fc IgG receptor. It was inferred that DAP IV represents a reliable marker for T-CLL derived from the OKT4 and Tmu positive subsets of T-lymphocytes, which encompasses the T-helper cell cohort.

Aged↗

PubMed1983

Simultaneous enzyme-immunocytochemical detection of antigens in monocellular specimens with monoclonal antibodies.

An immunoperoxidase and immunoalkaline phosphatase technique is described specially devised for separate cell specimens such as smears, cytospins, and tissue imprints. The reaction may be used alone or in combination for simultaneous staining with 2 different monoclonal antibodies. Monoclonal antibodies Leu3a, Leu2a, T4, T8, anti-kappa, and anti-lambda light chain were used in combination to visualize subsets of human lymphocytes selectively. Six cases of chronic lymphocytic leukemia were classified by this method.

Alkaline Phosphatase↗

PubMed1983

[Clinical picture and prognosis of generalized mastocytosis].

Proliferation of tissue mast cells in more than one organ is a rare disease, called generalized mastocytosis. Findings obtained in 35 cases were compared with 125 case reports in order to elucidate the course of the disease and its clinical picture. The results indicated that generalized mastocytosis has to be divided into two variants, designated systemic mastocytosis and malignant mastocytosis. Systemic mastocytosis is characterized by urticaria pigmentosa-like skin eruptions, with simultaneous infiltration of at least one visceral organ (usually bone marrow). Other frequently involved organs are spleen, liver, and lymph nodes. The age curve is biphasic, with one peak in early childhood and another in the 6th decade. The prognosis of systemic mastocytosis is generally favorable (actuarial survival 0.88 one year after diagnosis). In contrast to systemic mastocytosis, malignant mastocytosis does not show urticaria pigmentosa-like skin lesions. In this variant of generalized mastocytosis, frequently involved organs are bone marrow, spleen, liver, and lymph nodes. Common symptoms are anemia and eosinophilia. The age curve shows a peak in later life (6th and 7th decades). Malignant mastocytosis is never seen in children.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1983

Monocytic origin of the human hematopoietic cell line U-937 and its convertibility to macrophages evidenced by isoenzyme mapping.

U-937 represents a well established permanent human hematopoietic cell line. Electron microscopical and enzyme cytochemical studies as well as the analysis of surface glycoproteins have provided ample evidence for the monocytic origin of U-937. Upon stimulation with the tumour promotor 12-O-tetradecanoylphorbol-13-acetate (TPA), U-937 cells evolve into macrophage-like cells with phagocytic capacities. Since human blood monocytes (BM) are characterized by five acid esterase (AcE; EC 3.1.1.6) isoenzymes which are cell-specific in terms of isoelectric points (pI) and antigenicity, attempts were made in the present study to identify identical isoenzyme patterns in non- and TPA-stimulated U-937 cells. BM, cultured BM, non- and TPA-stimulated U-937 cells, as well as samples of resident human peritoneal macrophages (PM) as clearly-defined functional derivatives of BM, were subjected to isoenzyme analysis using isoelectric focusing (IEF). The five monocyte specific isoenzymes of AcE were identified in both populations of U-937. TPA-stimulated samples showed two additional bands, identical to those appearing in cultured BM after 4 days of glass-adherence and characteristic of resident human PM. Antisera raised against AcE of BM immunoprecipitated the two additional isoenzymes of TPA-stimulated U-937. It is concluded (1) the isoenzyme mapping of AcE documents the monocytic origin of U-937. (2) TPA-stimulation caricatures transformation of BM into resident tissue macrophages as far as pure morphology and AcE isoenzyme patterns are concerned. Thus, AcE isoenzyme mapping is apt for establishing reproducible and standardized criteria of different activation/differentiation states within the monocyte-macrophage lineage.

Animals↗

PubMed1983

Phenotypic heterogeneity of leukemic Sézary cells.

Leukemic blood cells from eight patients with Sézary's syndrome were analyzed for enzyme cytochemical features, Fc receptors, and surface phenotype. Enzyme cytochemically the cases were heterogeneous in their activity of acid esterase, acid phosphatase and dipeptidylaminopeptidase IV (DAP IV). Only one case showed positive staining for DAP IV. The expression of Fc receptors for IgG and IgM also varied. The DAP IV-positive case exhibited Fc mu receptors. In contrast, four other cases showed only Fc gamma receptors. Analysis of the surface antigen pattern of Sézary cells with monoclonal antibodies revealed the phenotype of helper T lymphocytes (Leu-3 a/OKT4+) in all but one case. The Leu-3 a/OKT4-negative case showed a phenotypic feature of natural killer cells (Leu-7+). The results obtained with the antibodies TU14 and Anti-human Lyt-1 were more heterogeneous. The heterogeneity of Sézary cells may be interpreted as a sign of differences in functional differentiation or of proliferation of different T-cell subclones.

Acid Phosphatase↗