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Biomedical subjects

G Damiani

Publications and source records attributed to G Damiani.

At least 91 records · Page 5Linked to original sources
PubMed1984

Progesterone enhances reactive oxygen intermediates production by cultured human monocytes.

Progesterone at placental tissue concentrations (5-20 micrograms/ml) markedly increases in vitro PMA-stimulated O-2 and H2O2 production by human cultured monocytes. This appears to contrast the well-known suppressive action of the hormone on the other cell mediated defence mechanisms. We suggest that these findings could cast a new light on progesterone's multiple and differentiated functions in the placental environment.

Cells, Cultured↗
PubMed1984

Binding of monoclonal antibody to cathepsin M located on the external surface of rabbit lysosomes.

A monoclonal antibody raised against rabbit liver cathepsin M binds to intact rabbit liver lysosomes. The binding is specific and is abolished by treating the lysosomes with trypsin, which has previously been shown to digest the membrane-bound cathepsin M [S. Pontremoli, E. Melloni, M. Michetti, F. Salamino, B. Sparatore, and B. L. Horecker (1982) Biochem, Biophys. Res. Commun. 106, 903-909]. Rabbit liver lysosomes are adsorbed onto Sepharose 4B coupled to anti-cathepsin M, but not to Sepharose 4B itself or to Sepharose coupled to a nonspecific antibody. The results confirm the location of membrane-bound cathepsin M on the outer surface of the lysosomal membrane.

Animals↗
PubMed1984

Pulmonary sarcoidosis: excess of helper T lymphocytes and T cell subset imbalance at sites of disease activity.

Different lymphocyte subpopulations have been evaluated in bronchoalveolar fluid and blood obtained from six patients with active and six with inactive pulmonary sarcoidosis and from six normal subjects by means of two recently described monoclonal antibodies, 5/9 and MLR4. The percentages of OKT4 positive (helper) and OKT8 positive (suppressor) T cells were also determined. Patients with active sarcoidosis had significantly higher proportions of 5/9 positive T cells in the bronchoalveolar fluid than patients with inactive disease (p less than 0.01) or normal subjects (p less than 0.001). In contrast, the proportions of 5/9 positive blood T cells were similar in the three groups studied. Patients with active sarcoidosis had also a greater proportion proportion of MLR4 positive T lymphocytes in bronchoalveolar fluid than patients with inactive disease or normal subjects (p less than 0.01 for each comparison), but similar proportions of MLR4 positive blood T cells were found in each group. The ratio of 5/9 positive to MLR4 positive T cells was higher in the bronchoalveolar fluid (but not in the blood) in patients with either active or inactive sarcoidosis than in normal subjects. These observations suggest that the MLR4 negative fraction rather than the MLR4 positive fraction of the 5/9 positive T cells is preferentially expanded in the lungs of patients with pulmonary sarcoidosis and may indicate a secondary role for the MLR4 positive T cells in producing lung injury in this disorder. Comparisons of the OKT4 positive and 5/9 positive T cells showed that in patients with active disease most of the lung T lymphocytes expressed both the OKT4 and the 5/9 surface antigens, so the 5/9 monoclonal antibody may be considered a good marker of activity in this disorder. Pulmonary sarcoidosis may be characterised by the preferential expansion of helper T cell subsets at sites of disease activity.

Adult↗
PubMed1983

A monoclonal antibody to platelet glycoproteins IIb and IIIa complex: its use in purifying human megakaryocytes from sternal bone marrow aspirates for immunofluorescence studies of Ia-like antigens.

A monoclonal antibody (PBM 6.4) to platelet and megakaryocyte glycoproteins IIb and IIIa has been obtained and used to purify human megakaryocytes from sternal bone marrow aspirates by a simple method, consisting of a Percoll gradient centrifugation followed by affinity adherence on PBM 6.4-coated plastic surface, "panning." Megakaryocytes, 80-90% pure and morphologically well preserved, were attached to poly-L-ornithine-coated multi-well microscope slides and immunofluorescence was done using monoclonal antibodies to human Ia-like antigens (DR, DC1). A higher proportion of DR-positive megakaryocytes was found, in comparison to the values reported by others, while DC1 antigen was detected on about 20% of megakaryocytes. The method described has the unique feature of enriching cells of the human megakaryocytic lineage from simple diagnostic sternal aspirates in an amount adequate for immunofluorescent and morphological analysis.

Animals↗
PubMed1982

Sequence analysis of heteropolymeric DNA synthesized in vitro by the enzyme terminal deoxynucleotidyl transferase and cloned in Escherichia coli.

We have synthesized in vitro single strands of DNA in reaction mixtures containing the terminal deoxynucleotidyl transferase (Bollum's enzyme), an oligo-dG as primer, the four common deoxynucleoside triphosphates and both Mg and Co ions. The resulting heteropolymers have been converted into double strands, tailed with oligo-dC sequences, annealed with an oligo-dG tailed plasmid vector and cloned in E. coli. Six recombinant plasmids have been isolated and characterized. Two of them have been sequenced. The heteropolymeric chains produced by the terminal transferase, ranging in size between 200 and 400 nucleotides, are richer in purines than in pyrimidines, except in the last portions. Open reading frames for 7-20 amino acids with repeated, in phase translational stop codons are present in these sequences and in their complements.

Base Sequence↗
PubMed1982

In vitro production of IgE by human peripheral blood mononuclear cells. III. Demonstration of a circulating IgE-bearing cell involved in the spontaneous IgE biosynthesis.

The presence of surface membrane IgE (SmIgE)-bearing cells in the peripheral blood (PB) of atopic patients was investigated by the use of isotype-specific rosettes of human red blood cells coupled to immunosorbent-purified rabbit or monoclonal mouse antibodies against human IgE (R or M anti-epsilon-HRBC). After dissociation of cell bound IgE by treatment with acid buffer, 2.1 +/- 0.3% and 1.2 +/- 0.3% circulating non-T, non-phagocytic, cells from atopic patients were still capable of forming rosettes with R or M anti-epsilon-HRBC, respectively. IgE molecules detectable on cells after dissociation of cytophilic IgE were quite resistant, like surface membrane IgM (SmIgM), to treatment with proteolytic enzymes, but they were removed under capping conditions by soluble anti-IgE antisera. All SmIgE-bearing (IgE+) cells also bore DR determinants, but many of them lacked SmIgM. Depletion of IgE+ cells strongly reduced the ability of PB lymphocyte suspensions from atopic patients to produce spontaneously IgE protein in vitro. Likewise, depletion of cells bearing DR determinants (DR+ cells) resulted in a marked decrease of the spontaneous IgE biosynthesis, whereas depletion of SmIgM-bearing (IgM+) cells had no effect. These data suggest that cells mainly implicated in the spontaneous IgE production in vitro seen in atopic patients are DR+ IgE+ IgM- circulating lymphocytes.

Antibodies, Monoclonal↗
PubMed1982

Human T cell subpopulations defined by a monoclonal antibody. I. A small subset is responsible for proliferation to allogeneic cells or to soluble antigens and for helper activity for B cell differentiation.

A monoclonal antibody (5/9) was obtained that reacts with the majority of alloactivated human T cells but only with 15 to 20% peripheral T cells. The populations reactive with the 5/9 antibody were separated from the remaining T cells by rosetting techniques using 5/9-coated ORBC. 5/9+ and 5/9- populations were analyzed for different in vitro activities. The helper activity of PWM- driven B cell differentiation appeared to be restricted and highly enriched in the 5/9+ population. In addition, 5/9+ cells contained all the cells capable of proliferation to tetanus toxoid and allogeneic cells. Thus, in vitro activities commonly used for evaluation of T cell function seem to be confined to a small fraction of peripheral T cells.

Animals↗
PubMed1981

Surface antigens specifically expressed by activated T cells in humans.

By immunizing BALB/c mice with human T cells derived from a secondary mixed lymphocyte reaction (MLR) and successive fusion of the spleen cells with P3X63Ag8U1 myeloma cells, several monoclonal antibodies to activated T cells were obtained. Four of them (MLR 1-4) were shown to be specific for activated T cells only, showing no reactivity with peripheral blood cells. The antigens recognized by these antibodies are differently distributed and appear on T cells at different times after stimulation. Thus, MLR2 stains nearly all the responder T cells present in a secondary MLR, while MLR 1, 3 and 4 stain only a fraction. MLR2 and 3 are also present on mitogen-stimulated cells. At least 3 of these MLR antigens may define different sets of activated T cells, and are likely to become useful in monitoring T cell activation also in the course of diseases involving the immune system.

Animals↗
PubMed1981

Analysis of HLA-DR polymorphism by two-dimensional peptide mapping.

Two-dimensional peptide mapping was used to study the polymorphism of DR antigens, membrane glycoproteins composed of two chains, alpha and beta, and encoded by the human major histocompatibility complex (MHC). Four DR antigens were purified by immunoabsorption from four human lymphoblastoid cell lines homozygous at the DR locus. After labeling with 125I, alpha and beta chains were separated by polyacrylamide gel electrophoresis in sodium dodecyl sulfate and digested with pepsin. Comparison of the peptide maps showed a marked degree of polymorphism among beta chains: only 43% of peptides were common to all four chains and 15-21% of the spots were unique to a given chain. By contrast, only a limited variability was observed among alpha chains. Homology was 75% for the four chains and the percentage of unique peptides was very low. DR7 did not possess even a single unique peptide. The limited variability among alpha chains and the lack of "private" peptides in one of them point to the conclusion that the beta chain is the unique carrier of the alloantigenic specificities. Higher homology within the known crossreactive groups was not observed, suggesting that the determinants responsible for crossreactivity are on different molecules. From a genetic point of view, because beta chains show allele-associated polymorphism, they are likely to be MHC encoded, whereas the minor differences among alpha chains do not allow a similar conclusion. The available data point to an analogy between these DR antigens and the mouse I-E/C antigens.

Alleles↗
PubMed1980

Macrophage variants in oxygen metabolism.

Whereas phagocytic cells from normal individuals have the capacity to kill ingested bacteria and parasites, those from patients with several uncommon genetic deficiency diseases are known to be defective in bactericidal activity. Studies on neutrophils of these patients have revealed fundamental defects in their ability to reduce molecular oxygen and metabolize it to superoxide anion, hydrogen peroxide, and oxygen radicals. In the present experiments, we describe a clone of a continuous murine macrophage-like cell line, J774.16, that, upon appropriate stimulation, activates the hexose monophosphate shunt, and produces superoxide anion and hydrogen peroxide. With nitroblue tetrazolium to select against cells capable of being stimulated by phorbol myristate acetate to reduce the dye to polymer--formazan--which is toxic fot cells, we have selected for variants that are defective in oxygen metabolism. Four of these subclones have been characterized and found to be lacking in the ability (a) to generate superoxide anion, as measured by cytochrome c reduction; (b) to produce hydrogen peroxide, as measured by the ability to form complex I with cytochrome c peroxidase; and (c) to be stimulated to oxidize glucose via the hexose monophosphate shunt. These variants appear to represent a useful model for studying the molecular basis for macrophage cytocidal activity.

Animals↗
PubMed1978

Disappearance of oxytocin-induced uterine tiredness by treatment with fructose-1,6-diphosphate. Experimental evidence.

Fructose-1,6-diphosphate (FDP) is able to abolish oxytocin induced spastic inertia in rat uterus. The clinical use of FDP is suggested by observation carried out on 30 deliveries with oxytocin induction. The FDP-treated patients (5 g of FDP in 50 ml of water by intravenous infusion) showed a statistically significant decrease of time elapsed between the beginning of uterine inertia and the recovery of uterine contractions (176 +/- 25.4 min) compared to controls (562 +/- 32.5 min).

Adolescent↗
PubMed1976

Synthesis of immunoglobulin by substrate attached mouse myeloma cells.

Cultured mouse myeloma cells grow in suspension and synthesize and secrete large amounts of immunoglobulin. Mouse myeloma cells which attach to a plastic substratum have been obtained by mutagenesis and subsequent selection. Normal mouse myeloma cells will also attach to plastic tissue culture dishes pre-treated with poly-L-lysine. The attached cells synthesize and secrete the same large amounts of immunoglobulin as the suspended cells.

Acridines↗