80 results on '"Battelli, M. G."'
Search Results
2. Interaction of volkensin with HeLa cells: binding, uptake, intracellular localization, degradation and exocytosis
- Author
-
Battelli, M. G., Musiani, S., Buonamici, L., Santi, S., Riccio, M., Maraldi, N. M., Girbés, T., and Stirpe, F.
- Published
- 2004
- Full Text
- View/download PDF
3. Hepatotoxicity of ricin, saporin or a saporin immunotoxin: xanthine oxidase activity in rat liver and blood serum
- Author
-
Battelli, M. G., Buonamici, L., Polito, L., Bolognesi, A., and Stirpe, F.
- Published
- 1996
- Full Text
- View/download PDF
4. T lymphocyte killing by a xanthine-oxidase-containing immunotoxin
- Author
-
Battelli, M. G., Abbondanza, A., Tazzari, P. L., Bolognesi, A., Lemoli, R. M., and Stirpe, F.
- Published
- 1992
- Full Text
- View/download PDF
5. Toxicity of xanthine oxidoreductase to malignant B lymphocytes
- Author
-
Battelli, M. G., Polito, L., Falà, F., Musiani, S., PIER LUIGI TAZZARI, Stirpe, F., Bolognesi, A., Battelli M.G., Polito L., Falà F., Musiani S., Tazzari P.L., Stirpe F., and Bolognesi A.
- Subjects
B-Lymphocytes ,Xanthine Oxidase ,Lymphoma, B-Cell ,L-Lactate Dehydrogenase ,Immunotoxins ,Apoptosis ,Mice ,Necrosis ,Cell Line, Tumor ,XANTHINE OXIDOREDUCTASE ,Animals ,Humans ,B CELL LYMPHOMA ,IMMUNOTOXIN ,Reactive Oxygen Species - Abstract
Reactive oxygen species (ROS) generated by xanthine oxidoreductase (XOR) were toxic to B lymphoma-derived Raji cells (positive for 8A monoclonal antibody, mAb). The sensitivity of these malignant cells to the hypoxanthine/XOR system was higher than that observed in peripheral human lymphocytes. The understanding of the mechanisms of cytotoxicity induced by XOR-produced ROS is essential in view of a possible clinical application. Cell death mostly had the feature of apoptosis and post-apoptotic necrosis and depended on the activity of XOR. Catalase, but not superoxide dismutase, protected cells from the toxicity of XOR, thus indicating that cell damage depended on the production of hydrogen peroxide. The toxicity of ROS was selectively targeted to malignant Raji cells by antibody-XOR conjugation, either directly, with an 8A-XOR conjugate, or indirectly, with an 8A mAb plus an anti-mouse IgG-XOR. Both direct and indirect immunotoxins induced apoptotic death to target cells in a dose-dependent manner. These conjugates showed no aspecific cytotoxicity in conditions very similar to the ex vivo treatment of cell suspension for bone marrow transplantation. Moreover, the prevalence of apoptotic death over necrosis may reduce the in vivo inflammatory response and its local and systemic consequences, thus becoming relevant in the construction of immunotoxins with therapeutic potential.
- Published
- 2006
6. Blood clearance and organ distribution and tissue concentration of native, homopolymerized and IgG-conjugated ribosome-inactivating proteins
- Author
-
Barbieri, L., primary, Battelli, M. G., additional, and Stirpe, F., additional
- Published
- 1990
- Full Text
- View/download PDF
7. Ricin toxicity to microglial and monocytic cells
- Author
-
Battelli, M. G., Musiani, S., Monti, B., Buonamici, L., Sparapani, M., Contestabile, A., and Stirpe, F.
- Published
- 2001
- Full Text
- View/download PDF
8. Determination of xanthine oxidase in human serum by a competitive enzyme-linked immunosorbent assay (ELISA)
- Author
-
Battelli, M. G., Abbondanza, A., Musiani, S., Buonamici, L., Strocchi, P., Tazzari, P. L., Gramantieri, L., and Stirpe, F.
- Published
- 1999
- Full Text
- View/download PDF
9. Simulated ischaemia-reperfusion conditions increase xanthine dehydrogenase and oxidase activities in rat brain slices
- Author
-
Battelli, M. G., Buonamici, L., Virgili, M., Abbondanza, A., and Contestabile, A.
- Published
- 1998
- Full Text
- View/download PDF
10. Purification and properties of a new glutathione-dependent thiol:disulphide oxidoreductase from rat liver
- Author
-
Battelli, M G and Lorenzoni, E
- Abstract
A new GSSG-dependent thiol:disulphide oxidoreductase was extensively purified from rat liver cytosol. The enzymic protein shows molecular weight 40 000 as determined by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, and 43 000 as determined by thin-layer gel filtration on Bio-Gel P-100. The pI is 8.1. This enzyme converts rat liver xanthine dehydrogenase into an oxidase, in the presence of oxidized glutathione. Other disulphide compounds are either inactive or far less active than oxidized glutathione in the enzymic oxidation of rat liver xanthine dehydrogenase. The enzyme also catalyses the reduction of the disulphide bond of ricin and acts as a thioltransferase and as a GSH:insulin transhydrogenase. The enzymic activity was measured in various organs of newborn and adult rats.
- Published
- 1982
- Full Text
- View/download PDF
11. Bryodin, a ribosome-inactivating protein from the roots of Bryonia dioica L. (white bryony)
- Author
-
Stirpe, F, Barbieri, L, Battelli, M G, Falasca, A I, Abbondanza, A, Lorenzoni, E, and Stevens, W A
- Abstract
Bryodin is a strongly basic (pI greater than or equal to 9.5) glycoprotein (neutral sugar content 6.3%) with Mr 30,000, purified from the roots of Bryonia dioica (white bryony). This protein inhibits protein synthesis by a rabbit reticulocyte lysate with and ID50 (concentration causing 50% inhibition) of 0.12 nM (3.6 ng/ml) and has much less effect on protein synthesis by whole cells, with ID50 values ranging from 46 nM to 2.27 microM (1.4-67 micrograms/ml). Bryodin acts by inactivating ribosomes, with a less-than-equimolar ratio, which suggests a catalytic action. Bryodin decreases the number of local lesions induced by tobacco mosaic virus in the leaves of Nicotiana glutinosa. From all its properties, bryodin can be considered to be a ribosome-inactivating protein, similar to those already known [reviews: Barbieri & Stirpe (1982) Cancer Surveys 1, 489-520; Stirpe & Barbieri (1986) FEBS Lett. 195, 1-8].
- Published
- 1986
- Full Text
- View/download PDF
12. Milk xanthine oxidase type D (dehydrogenase) and type O (oxidase). Purification, interconversion and some properties
- Author
-
Battelli, M. G., Lorenzoni, E., and Stirpe, F.
- Abstract
1. The xanthine oxidase of cow's milk, crude or purified, appears as an oxidase (type O), and can be converted almost completely into a NAD+-dependent dehydrogenase (type D) by treatment with dithioerythritol or dihydrolipoic acid, but only to a small extent by other thiols. 2. The D form of the enzyme is inhibited by NADH, which competes with NAD+. 3. The kinetic constants of the two forms of the enzyme are similar to those of the corresponding forms of rat liver xanthine oxidase. 4. Milk xanthine oxidase is converted into an irreversible O form by pretreatment with chymotrypsin, papain or subtilisin, but only partially with trypsin. 5. The enzyme as purified shows a major faster band and a minor slower band on gel electrophoresis. The slower band is greatly reinforced after xanthine oxidase is converted into the irreversible O form by chymotrypsin.
- Published
- 1973
- Full Text
- View/download PDF
13. Selective cytotoxicity of an oxygen-radical-generating enzyme conjugated to a monoclonal antibody
- Author
-
Battelli, M. G., Abbondanza, A., PIER LUIGI TAZZARI, Dinota, A., Rizzi, S., Grassi, G., Gobbi, M., and Stirpe, F.
- Subjects
Oxygen ,Xanthine Oxidase ,Free Radicals ,Cell Survival ,Immunotoxins ,Tumor Cells, Cultured ,Antibodies, Monoclonal ,Humans ,Hematopoietic Stem Cells ,Cell Line ,Research Article - Abstract
The monoclonal antibody 8A, which recognizes a human plasma cell-associated antigen, was covalently linked to xanthine oxidase in a conjugate maintaining both immunological and enzymatic properties. A significant degree of target cell lysis was obtained at an enzyme concentration that was ineffective on non-target cells and on myeloid staminal cells (CFU-GM). The cytotoxic activity was abolished by an excess of antibody, by allopurinol and by superoxide dismutase and catalase. A possible use of the conjugate for bone marrow purging in multiple myeloma patients is suggested.
- Published
- 1988
14. CD38 as a target of IB4 mAb carrying saporin-S6: Design of an immunotoxin for ex vivo depletion of hematological CD38+ neoplasia
- Author
-
Bolognesi, A., Letizia Polito, Farini, V., Bortolotti, M., Tazzari, P. L., Ratta, M., Ravaioli, A., Horenstein, A. L., Stirpe, F., Battelli, M. G., and Malavasi, F.
15. CD38 as a target of IB4 mAb carrying saporin-S6: Design of an immunotoxin for ex vivo depletion of hematological CD38+ neoplasia
- Author
-
Andrea Bolognesi, Polito, L., Farini, V., Bortolotti, M., Tazzari, P. L., Ratta, M., Ravaioli, A., Horenstein, A. L., Stirpe, F., Battelli, M. G., Malavasi, F., Bolognesi A., Polito L., Farini V., Bortolotti M., Tazzari P.L., Ratta M., Ravaioli A., Horenstein A.L., Stirpe F., Battelli M.G., and Malavasi F.
- Subjects
SAPORIN-S6 ,Cell Separation ,In Vitro Techniques ,immune system diseases ,hemic and lymphatic diseases ,Cell Line, Tumor ,Humans ,IMMUNOTOXIN ,IMMUNOTHERAPY ,N-Glycosyl Hydrolases ,Plant Proteins ,Protein Synthesis Inhibitors ,therapy ,Immunotoxins ,toxins ,Antibodies, Monoclonal ,hemic and immune systems ,monoclonal antibodies ,ADP-ribosyl Cyclase 1 ,Antineoplastic Agents, Phytogenic ,Saporins ,Drug Design ,Hematologic Neoplasms ,IB4 ,Ribosome Inactivating Proteins, Type 1 ,CD38 - Abstract
An anti-CD38 mAb (IB4) coupled to saporin-S6, a type 1 ribosome-inactivating protein (RIP), was designed for ex vivo or loco-regional therapeutical applications in myeloma and lymphoma. The ability of this immunotoxin to eliminate CD38+ cells was studied in vitro on selected CD38+ human cell lines (Raji, HBL6, L540 and CEM) and on CD38+ neoplastic cells from a Non Hodgkin Lymphoma (NHL) patient. HBL6, Raji and L540 cells resulted very sensitive to the IB4/saporin-S6 conjugate, concentrations as low as 100 pM of the immunotoxin completely inhibiting protein synthesis. CD38+ neoplastic cells from the NHL patient were completely eliminated after treatment with immunotoxin at 10 nM concentration. CFU-c rescue by bone marrow precursors was maintained after exposure to the immunotoxin. These results indicate that IB4/saporin-S6 is endowed with strong and specific cytotoxic effects on selected CD38+ tumor cells lineages. Consequently, it is reasonable to propose a clinical use of the IB4/saporin-S6 for ex vivo purging of unwanted cells (e.g. depletion of contaminating neoplastic cells in aphereses obtained from G-CSF-treated patients) or for loco-regional therapies of CD38+ tumors.
16. Xanthine oxidase type D (dehydrogenase) in the intestine and other organs of the rat
- Author
-
Battelli, M G, primary, Corte, E D, additional, and Stirpe, F, additional
- Published
- 1972
- Full Text
- View/download PDF
17. Excitotoxic increase of xanthine dehydrogenase and xanthine oxidase in the rat olfactory cortex
- Author
-
Battelli, M. G., Buonamici, L., Abbondanza, A., and Virgili, M.
- Published
- 1995
- Full Text
- View/download PDF
18. New ribosome-inactivating proteins with polynucleotide:adenosine glycosidase and antiviral activities from Basella rubra L. and Bougainvillea spectabilis Willd
- Author
-
M. Giulia Bartelli, Letizia Polito, Paola Valbonesi, Fabiola Olivieri, M. Di Loreto, F. Stirpe, F. Del Vecchio Blanco, Augusto Parente, M. Vittoria Carusi, A. Di Maro, Luigi Barbieri, Andrea Bolognesi, Eugenio Benvenuto, Bolognesi, A., Polito, L., Olivieri, F., Barbieri, L., Battelli, M. G., Carusi, M. V., Benvenuto, E., DEL VECCHIO BLANCO, F., DI MARO, Antimo, Parente, A., DI LORETO, M., and Stirpe, F.
- Subjects
Male ,endocrine system ,endocrine system diseases ,Molecular Sequence Data ,Antiviral protein ,Ribosome Inactivating Proteins ,Plant Science ,Biology ,digestive system ,Antiviral Agents ,Basella ,Mice ,food ,Basella rubra ,Genetics ,Tumor Cells, Cultured ,Animals ,Humans ,Amino Acid Sequence ,N-Glycosyl Hydrolases ,Plant Proteins ,Protein Synthesis Inhibitors ,Bougainvillea ,Momordin ,Ribosome-inactivating protein ,Polynucleotide:adenosine glycosidase ,3T3 Cells ,DNA ,Bougainvillea spectabilis ,Ribosomal RNA ,Plants ,biology.organism_classification ,food.food ,Rats ,RNA, Bacterial ,Biochemistry ,RNA, Viral ,Female ,Rabbits ,Poly A ,Ribosomes ,HeLa Cells - Abstract
New single-chain (type 1) ribosome-inactivating proteins (RIPs) were isolated from the seeds of Basella rubra L. (two proteins) and from the leaves of Bougainvillea spectabilis Willd. (one protein). These RIPs inhibit protein synthesis both in a cell-free system, with an IC50 (concentration causing 50% inhibition) in the 10(-10) M range, and by various cell lines, with IC50S in the 10(-8)-10(-6) M range. All three RIPs released adenine not only from rat liver ribosomes but also from Escherichia coli rRNA, polyadenylic acid, herring sperm DNA, and artichoke mottled crinkle virus (AMCV) genomic RNA, thus being polynucleotide:adenosine glycosidases. The proteins from Basella rubra had toxicity to mice similar to that of most type 1 RIPs (Barbieri et al., 1993, Biochim Biophys Acta 1154: 237-282) with an LD50 (concentration that is 50% lethal)or = 8 mg.kg-1 body weight, whilst the RIP from Bougainvillea spectabilis had an LD5032 mg.kg-1. The N-terminal sequence of the two RIPs from Basella rubra had 80-93% identity, whereas it differed from the sequence of the RIP from Bougainvillea spectabilis. When tested with antibodies against various RIPs, the RIPs from Basella gave some cross-reactivity with sera against dianthin 32, and weak cross-reactivity with momordin I and momorcochin-S, whilst the RIP from Bougainvillea did not cross-react with any antiserum tested. An RIP from Basella rubra and one from Bougainvillea spectabilis were tested for antiviral activity, and both inhibited infection of Nicotiana benthamiana by AMCV.
- Published
- 1997
19. Purification and partial characterization of single-chain ribosome-inactivating proteins from the seeds of Phytolacca dioica L
- Author
-
Augusto Parente, Ada Abbondanza, Fiorenzo Stirpe, Gesualdo Siniscalco Gigliano, Pier Luigi Tazzari, Paolo De Luca, Manuela J.W. Sande, Luigi Barbieri, Andrea Bolognesi, Maria Giulia Battelli, Parente, A., De Luca, P., Bolognesi, A., Barbieri, L., Battelli, M. G., Abbondanza, A., Sande, M. J. W., SINISCALCO GIGLIANO, Gesualdo, Tazzari, P. L., and Stirpe, F.
- Subjects
endocrine system ,Glycoside Hydrolases ,endocrine system diseases ,Molecular Sequence Data ,Biophysics ,Phytolacca dioica ,digestive system ,Biochemistry ,Ribosome ,Cell Line ,Mice ,chemistry.chemical_compound ,Structural Biology ,Phytolacca americana ,Genetics ,Protein biosynthesis ,Animals ,Amino Acid Sequence ,N-Glycosyl Hydrolases ,Plant Proteins ,chemistry.chemical_classification ,biology ,Immunotoxins ,Ribosome-inactivating protein ,biology.organism_classification ,Saporins ,Molecular biology ,Phytolaccaceae ,Rats ,Ricin ,chemistry ,Protein Biosynthesis ,Seeds ,Ribosome Inactivating Proteins, Type 1 ,Female ,Rabbits ,Glycoprotein ,Ribosomes - Abstract
Three ribosome-inactivating proteins (RIPs) similar to those already known (Stirpe et al. (1992) Bio/Technology 10, 405–412) were purified from the seeds of Phytolacca dioica . These proteins, called Phytolacca dioica RIPs (PD-S1, PD-S2 and PD-S3 RIPs), are glycoproteins, with M r approx. 30 000, inhibit protein synthesis by a rabbit reticulocyte lysate and phenylalanine polymerization by isolated ribosomes, and depurinate rat liver rRNA in an apparently identical manner as the A-chain of ricin and other RIPs (Endo et al. (1987) J. Biol. Chem. 262, 5908–5912). Part of the purified rat liver ribosomes appeared resistant to the action of PD-S RIPs. The most abundant protein, PD-S2 RIP, gave a weak or nil cross-reaction with sera against various other RIPs, including a pokeweed antiviral protein from the roots of Phytolacca americana . PD-S2 RIP was linked to a monoclonal antibody (Ber-H2) against the CD30 human lymphocyte antigen and the resulting immunotoxin was selectively toxic to the CD30+ Hodgkin's lymphoma-derived L540 cell line.
- Published
- 1993
20. Endocytosis and intracellular localisation of type 1 ribosome-inactivating protein saporin-s6.
- Author
-
Bolognesi A, Polito L, Scicchitano V, Orrico C, Pasquinelli G, Musiani S, Santi S, Riccio M, Bortolotti M, and Battelli MG
- Subjects
- DNA Damage, Endosomes metabolism, Fluorescent Antibody Technique, Indirect, Golgi Apparatus metabolism, HeLa Cells metabolism, Humans, Microscopy, Confocal, Microscopy, Electron, Transmission, Protein Synthesis Inhibitors pharmacokinetics, Ribosome Inactivating Proteins, Type 1 pharmacokinetics, Saporins, Endocytosis, Ribosome Inactivating Proteins, Type 1 metabolism
- Abstract
Saporin-S6 is a single-chain ribosome-inactivating protein (RIP) that has low toxicity in cells and animals. When the protein is bound to a carrier that facilitates cellular uptake, the protein becomes highly and selectively toxic to the cellular target of the carrier. Thus, saporin-S6 is one of the most widely used RIPs in the preparation of immunoconjugates for anti-cancer therapy. The endocytosis of saporin-S6 by the neoplastic HeLa cells and the subsequent intracellular trafficking were investigated by confocal microscopy that utilises indirect immunofluorescence analysis and transmission electron microscopy that utilises a direct assay with gold-conjugated saporin-S6 and an indirect immunoelectron microscopy assay. Our results indicate that saporin-S6 was taken up by cells mainly through receptor-independent endocytosis. Confocal microscopy analysis showed around 30% co-localisation of saporin-S6 with the endosomal compartment and less than 10% co-localisation with the Golgi apparatus. The pathway identified by the immunofluorescence assay and transmission electron microscopy displayed a progressive accumulation of saporin-S6 in perinuclear vesicular structures. The main findings of this work are the following: i) the nuclear localisation of saporin-S6 and ii) the presence of DNA gaps resulting from abasic sites in HeLa nuclei after intoxication with saporin-S6.
- Published
- 2012
21. Increased serum level of xanthine oxidoreductase in liver transplanted patients.
- Author
-
Battelli MG, Ravaioli M, Musiani S, Scicchitano V, Grazi GL, and Bolognesi A
- Subjects
- Adult, Aged, Enterocytes metabolism, Enzyme-Linked Immunosorbent Assay, Female, Humans, Male, Middle Aged, Time Factors, Liver metabolism, Liver Transplantation, Reperfusion, Transplants, Xanthine Dehydrogenase blood
- Abstract
Xanthine oxidoreductase (XOR) leakage into serum has been observed in various types of liver pathology as well as after liver transplantation (LT). We determined the amount of XOR associated with LT to investigate the changes in serum enzyme level during the LT procedure and the post-operative period. Additionally, we examined whether there was any correlation between XOR levels and the surgical technique. XOR levels were measured by a competitive ELISA. In a first group of patients, the portal vein was flushed before the liver and systemic reperfusions, which occurred simultaneously. In the second group, the graft was flushed with blood from the portal vein before the systemic reperfusion. XOR showed a marked elevation in the caval effluent collected during LT and was higher compared to control serum levels at all time points that were examined after LT. The XOR levels during LT were also higher than samples taken pre-LT or from the portal blood flush before reperfusion. The XOR level was higher in Group 2 than in Group 1. Enhancement of the XOR serum level during LT was not derived from enterocytes, and it should be attributed to enzyme leakage from graft liver cells. We report the elevation of serum XOR during the three weeks following LT for the first time, as well as the influence of the graft reperfusion technique on XOR serum levels.
- Published
- 2011
22. Toxicity of xanthine oxidoreductase to malignant B lymphocytes.
- Author
-
Battelli MG, Polito L, Falà F, Musiani S, Tazzari PL, Stirpe F, and Bolognesi A
- Subjects
- Animals, Apoptosis drug effects, B-Lymphocytes drug effects, B-Lymphocytes pathology, Cell Line, Tumor, Humans, Immunotoxins metabolism, Immunotoxins toxicity, L-Lactate Dehydrogenase metabolism, Lymphoma, B-Cell enzymology, Lymphoma, B-Cell pathology, Lymphoma, B-Cell therapy, Mice, Necrosis, Reactive Oxygen Species metabolism, Reactive Oxygen Species toxicity, B-Lymphocytes enzymology, Xanthine Oxidase metabolism
- Abstract
Reactive oxygen species (ROS) generated by xanthine oxidoreductase (XOR) were toxic to B lymphoma-derived Raji cells (positive for 8A monoclonal antibody, mAb). The sensitivity of these malignant cells to the hypoxanthine/XOR system was higher than that observed in peripheral human lymphocytes. The understanding of the mechanisms of cytotoxicity induced by XOR-produced ROS is essential in view of a possible clinical application. Cell death mostly had the feature of apoptosis and post-apoptotic necrosis and depended on the activity of XOR. Catalase, but not superoxide dismutase, protected cells from the toxicity of XOR, thus indicating that cell damage depended on the production of hydrogen peroxide. The toxicity of ROS was selectively targeted to malignant Raji cells by antibody-XOR conjugation, either directly, with an 8A-XOR conjugate, or indirectly, with an 8A mAb plus an anti-mouse IgG-XOR. Both direct and indirect immunotoxins induced apoptotic death to target cells in a dose-dependent manner. These conjugates showed no aspecific cytotoxicity in conditions very similar to the ex vivo treatment of cell suspension for bone marrow transplantation. Moreover, the prevalence of apoptotic death over necrosis may reduce the in vivo inflammatory response and its local and systemic consequences, thus becoming relevant in the construction of immunotoxins with therapeutic potential.
- Published
- 2005
23. CD38 as a target of IB4 mAb carrying saporin-S6: design of an immunotoxin for ex vivo depletion of hematological CD38+ neoplasia.
- Author
-
Bolognesi A, Polito L, Farini V, Bortolotti M, Tazzari PL, Ratta M, Ravaioli A, Horenstein AL, Stirpe F, Battelli MG, and Malavasi F
- Subjects
- Antibodies, Monoclonal pharmacology, Antineoplastic Agents, Phytogenic pharmacology, Cell Line, Tumor, Cell Separation, Drug Design, Humans, Immunotoxins pharmacology, In Vitro Techniques, N-Glycosyl Hydrolases pharmacology, Plant Proteins pharmacology, Protein Synthesis Inhibitors pharmacology, Ribosome Inactivating Proteins, Type 1, Saporins, ADP-ribosyl Cyclase 1 metabolism, Hematologic Neoplasms immunology, Hematologic Neoplasms therapy, Immunotoxins immunology
- Abstract
An anti-CD38 mAb (IB4) coupled to saporin-S6, a type 1 ribosome-inactivating protein (RIP), was designed for ex vivo or loco-regional therapeutical applications in myeloma and lymphoma. The ability of this immunotoxin to eliminate CD38+ cells was studied in vitro on selected CD38+ human cell lines (Raji, HBL6, L540 and CEM) and on CD38+ neoplastic cells from a Non Hodgkin Lymphoma (NHL) patient. HBL6, Raji and L540 cells resulted very sensitive to the IB4/saporin-S6 conjugate, concentrations as low as 100 pM of the immunotoxin completely inhibited protein synthesis. CD38+ neoplastic cells from the NHL patient were completely eliminated after treatment with immunotoxin at 10 nM concentration. CFU-c rescue by bone marrow precursors was maintained after exposure to the immunotoxin. These results indicate that IB4/saporin-S6 is endowed with strong and specific cytotoxic effects on selected CD38+ tumor cells lineages. Consequently, it is reasonable to propose a clinical use of the IB4/saporin-S6 for ex vivo purging of unwanted cells (e.g. depletion of contaminating neoplastic cells in aphereses obtained from G-CSF-treated patients) or for loco-regional therapies of CD38+ tumors.
- Published
- 2005
24. Oxidative stress to human lymphocytes by xanthine oxidoreductase activity.
- Author
-
Battelli MG, Musiani S, Tazzari PL, and Stirpe F
- Subjects
- Apoptosis, CD4-Positive T-Lymphocytes metabolism, CD8-Positive T-Lymphocytes metabolism, Catalase metabolism, DNA Fragmentation, Flow Cytometry, Humans, Hypoxanthine pharmacology, Lymphocytes drug effects, Superoxide Dismutase metabolism, Time Factors, Transglutaminases metabolism, Lymphocytes enzymology, Oxidative Stress, Oxidoreductases Acting on CH-NH Group Donors metabolism, Xanthine metabolism
- Abstract
The in vitro toxicity of the reactive oxygen species generating enzyme xanthine oxidoreductase (XOR) to human peripheral blood lymphocytes was studied after stimulation with phytohaemoagglutinin or anti-CD3/CD28 antibodies. Apoptosis and necrosis were induced by the XOR/hypoxanthine system in a time- and concentration-dependent manner. CD8+ lymphocytes showed a higher sensitivity than CD4+ cells to the XOR/hypoxanthine system. The occurrence of apoptosis was demonstrated by annexin-V binding to injured cell membrane, which was the most precocious alteration observed, followed by the increment of transglutaminase activity, which was significant at the lowest XOR concentration used. Nuclear damage was assessed by the increased hypodiploid nuclei and by DNA migration on gel electrophoresis, which turned to an apoptotic pattern before the occurrence of cell membrane necrotic lesions. Apoptosis was induced by XOR activity proportionally to substrate concentration and was prevented by the competitive enzyme inhibitor, allopurinol. The hydrogen peroxide scavenging enzyme, catalase, gave a higher protection than superoxide dismutase from the toxicity caused by the XOR/hypoxanthine system. Necrosis occurs in a variable percentage indicating that reactive oxygen species may trigger both apoptosis and necrosis in proliferating human lymphocytes, mostly depending on XOR concentration.
- Published
- 2001
- Full Text
- View/download PDF
25. Serum xanthine oxidase in human liver disease.
- Author
-
Battelli MG, Musiani S, Valgimigli M, Gramantieri L, Tomassoni F, Bolondi L, and Stirpe F
- Subjects
- Adult, Chronic Disease, Female, Humans, Male, Middle Aged, Cholestasis enzymology, Hepatitis, Viral, Human enzymology, Liver Cirrhosis enzymology, Xanthine Oxidase blood
- Abstract
Objectives: High concentrations of serum xanthine oxidase (XO) have been reported during human liver disease and hepatocyte injury in experimental settings. However, it is unclear whether this elevation reflects hepatocyte necrosis or has a different meaning., Methods: The serum level of XO in 64 patients with chronic liver disease (17 patients with cirrhosis, 30 with chronic hepatitis, and 17 with cholestatic disorders) and in 12 control subjects was determined by a competitive ELISA. Conventional serum markers of liver damage were assessed in all patients, and grading and staging were scored in the chronic hepatitis group according to Knodell., Results: The XO serum levels were significantly higher in the patients than in the controls. The differences were also significant when controls were compared to patients with chronic hepatitis and cholestatic disorders separately, but not when compared to the cirrhosis group. Patients with cholestatic disorders had XO values higher than those of patients with cirrhosis or chronic hepatitis. XO levels did not correlate with stage and grade in chronic hepatitis group. We found a weak but significant positive correlation in patients between XO serum level and gamma-glutamyl transpeptidase (r = 0.37). This correlation was stronger when chronic hepatitis (r = 0.42) and, especially cholestatic disorders (r = 0.71), were separately tested, but was absent in the cirrhosis group. The XO values positively correlated with alkaline phosphatase in patients with cholestatic disorders. A level of serum XO >32 microg/ml specifically identified cholestatic disorders in our study population., Conclusions: A marked elevation of serum XO in patients with chronic liver disease seems to reflect the presence of cholestasis. No correlation between XO levels and histological or serum evidence of hepatocyte necrosis was found in these patients.
- Published
- 2001
- Full Text
- View/download PDF
26. Different sensitivity of CD30+ cell lines to Ber-H2/saporin-S6 immunotoxin.
- Author
-
Battelli MG, Bolognesi A, Olivieri F, Polito L, and Stirpe F
- Subjects
- Antineoplastic Agents chemistry, Binding Sites, Cell Membrane metabolism, Drug Screening Assays, Antitumor, Exocytosis drug effects, Humans, Immunotoxins chemistry, Immunotoxins metabolism, Ki-1 Antigen analysis, Neoplasm Proteins biosynthesis, Phenotype, Ribosome Inactivating Proteins, Type 1, Saporins, Tumor Cells, Cultured, Antineoplastic Agents pharmacology, Immunotoxins pharmacology, Ki-1 Antigen chemistry, N-Glycosyl Hydrolases, Plant Proteins chemistry
- Abstract
The in vitro sensitivity of cells to a Ber-H2(anti-CD30)/saporin-S6 immunotoxin has been investigated. The CD30+ cell lines, K562, L428 and L540, were used to study cell binding, uptake and degradation of the immunotoxin. K562 cells were less sensitive than L428 and L540 cells to the immunotoxin by approximately one order of magnitude. The difference in cytotoxicity correlated with the intracellular accumulation and with the ratio of degraded over total internalized Ber-H2/saporin-S6, regardless of the immunotoxin binding to the cells. After 6 h incubation, the less sensitive K562 cells (i) accumulated only one third and one tenth of the immunotoxin accumulated by the more sensitive L428 and L540 cells, respectively, and (ii) degraded two thirds of the internalized protein versus one third degraded by either L428 or L540 cells. Ammonium chloride and chloroquine reduced the cytotoxicity of the immunotoxin towards K562 but not to L540 cells. This effect correlated with the increment of immunotoxin catabolism by K562 cells in the presence of chloroquine. In conclusion, uptake alone of an immunotoxin by target cells is not sufficient to assure its efficacy which might also depend on intracellular routing. Only a cytotoxicity test may be really predictive.
- Published
- 1998
- Full Text
- View/download PDF
27. New ribosome-inactivating proteins with polynucleotide:adenosine glycosidase and antiviral activities from Basella rubra L. and bougainvillea spectabilis Willd.
- Author
-
Bolognesi A, Polito L, Olivieri F, Valbonesi P, Barbieri L, Battelli MG, Carusi MV, Benvenuto E, Del Vecchio Blanco F, Di Maro A, Parente A, Di Loreto M, and Stirpe F
- Subjects
- 3T3 Cells, Amino Acid Sequence, Animals, Antiviral Agents chemistry, Antiviral Agents isolation & purification, DNA metabolism, Female, HeLa Cells, Humans, Male, Mice, Molecular Sequence Data, N-Glycosyl Hydrolases chemistry, N-Glycosyl Hydrolases isolation & purification, N-Glycosyl Hydrolases pharmacology, Plant Proteins chemistry, Plant Proteins isolation & purification, Plant Proteins pharmacology, Poly A metabolism, Protein Synthesis Inhibitors chemistry, Protein Synthesis Inhibitors isolation & purification, RNA, Bacterial metabolism, RNA, Viral metabolism, Rabbits, Rats, Ribosome Inactivating Proteins, Ribosomes, Tumor Cells, Cultured, Antiviral Agents pharmacology, N-Glycosyl Hydrolases metabolism, Plants enzymology, Protein Synthesis Inhibitors pharmacology
- Abstract
New single-chain (type 1) ribosome-inactivating proteins (RIPs) were isolated from the seeds of Basella rubra L. (two proteins) and from the leaves of Bougainvillea spectabilis Willd. (one protein). These RIPs inhibit protein synthesis both in a cell-free system, with an IC50 (concentration causing 50% inhibition) in the 10(-10) M range, and by various cell lines, with IC50S in the 10(-8)-10(-6) M range. All three RIPs released adenine not only from rat liver ribosomes but also from Escherichia coli rRNA, polyadenylic acid, herring sperm DNA, and artichoke mottled crinkle virus (AMCV) genomic RNA, thus being polynucleotide:adenosine glycosidases. The proteins from Basella rubra had toxicity to mice similar to that of most type 1 RIPs (Barbieri et al., 1993, Biochim Biophys Acta 1154: 237-282) with an LD50 (concentration that is 50% lethal) < or = 8 mg.kg-1 body weight, whilst the RIP from Bougainvillea spectabilis had an LD50 > 32 mg.kg-1. The N-terminal sequence of the two RIPs from Basella rubra had 80-93% identity, whereas it differed from the sequence of the RIP from Bougainvillea spectabilis. When tested with antibodies against various RIPs, the RIPs from Basella gave some cross-reactivity with sera against dianthin 32, and weak cross-reactivity with momordin I and momorcochin-S, whilst the RIP from Bougainvillea did not cross-react with any antiserum tested. An RIP from Basella rubra and one from Bougainvillea spectabilis were tested for antiviral activity, and both inhibited infection of Nicotiana benthamiana by AMCV.
- Published
- 1997
- Full Text
- View/download PDF
28. Toxicity of ricin and volkensin, two ribosome-inactivating proteins, to microglia, astrocyte, and neuron cultures.
- Author
-
Sparapani M, Buonamici L, Ciani E, Battelli MG, Ceccarelli G, Stirpe F, and Contestabile A
- Subjects
- Animals, Astrocytes cytology, Astrocytes metabolism, Cell Survival, Cells, Cultured, L-Lactate Dehydrogenase metabolism, Leucine metabolism, Leucine pharmacology, Microglia cytology, Microglia metabolism, Neurons cytology, Neurons metabolism, Rats, Rats, Wistar, Ribosome Inactivating Proteins, Type 2, Ribosomes, Astrocytes drug effects, Glycoproteins, Microglia drug effects, N-Glycosyl Hydrolases, Neurons drug effects, Plant Lectins, Plant Proteins toxicity, Protein Synthesis Inhibitors toxicity, Ricin toxicity, Toxins, Biological toxicity
- Abstract
Ricin and volkensin, two potent toxins belonging to the family of ribosome-inactivating proteins (RIPs), have been largely exploited in recent years in in vivo experiments of neuronal degeneration consequent to suicide transport or immunolesioning. We have determined both the toxicity of, and the inhibition of, protein synthesis by ricin and volkensin in in vitro cultures enriched in microglial cells, astrocytes, or neurons. In microglial cultures, 50% of toxicity (estimated by LDH released from dead cells) after 24 h exposure to RIPs was obtained with volkensin at 2.2x10(-12) M concentration and 50% of protein synthesis inhibition at 2x10(-14) M concentration. Both values were higher by about one order of magnitude in astrocyte-enriched cultures. Toxicity of, and inhibition of, protein synthesis by, ricin were lower for both cell types by about 1 order of magnitude as compared to volkensin. Cerebellar granule neurons in culture survived remarkably well to 24 h exposure to ricin or volkensin, although their protein synthesis was effectively inhibited by the two toxins with a potency similar to that found for astrocytes. These results demonstrate that glial cells, in particular microglia, are very sensitive to RIPs toxicity and should, therefore, be a primary target of these toxins when injected in vivo. Thus, the damage observed after in vivo experiments could be partly related to diffusion of toxic substances from early-affected glial cells.
- Published
- 1997
- Full Text
- View/download PDF
29. Ribosome-inactivating lectins with polynucleotide:adenosine glycosidase activity.
- Author
-
Battelli MG, Barbieri L, Bolognesi A, Buonamici L, Valbonesi P, Polito L, Van Damme EJ, Peumans WJ, and Stirpe F
- Subjects
- 3T3 Cells, Animals, Cell Line, Cell-Free System, Galanthus, HeLa Cells, Humans, Kinetics, Lectins pharmacology, Mice, Plant Lectins, Ribosome Inactivating Proteins, Lectins metabolism, Protein Synthesis Inhibitors pharmacology, Ribosomes metabolism
- Abstract
Lectins from Aegopodium podagraria (APA), Bryonia dioica (BDA), Galanthus nivalis (GNA), Iris hybrid (IRA) and Sambucus nigra (SNAI), and a new lectin-related protein from Sambucus nigra (SNLRP) were studied to ascertain whether they had the properties of ribosome-inactivating proteins (RIP). IRA and SNLRP inhibited protein synthesis by a cell-free system and, at much higher concentrations, by cells and had polynucleotide:adenosine glycosidase activity, thus behaving like non-toxic type 2 (two chain) RIP. APA and SNAI had much less activity, and BDA and GNA did not inhibit protein synthesis.
- Published
- 1997
- Full Text
- View/download PDF
30. Toxicity and cytotoxicity of nigrin b, a two-chain ribosome-inactivating protein from Sambucus nigra: comparison with ricin.
- Author
-
Battelli MG, Citores L, Buonamici L, Ferreras JM, de Benito FM, Stirpe F, and Girbés T
- Subjects
- Analysis of Variance, Animals, Binding, Competitive, HeLa Cells cytology, HeLa Cells metabolism, Humans, Lethal Dose 50, Mice, N-Glycosyl Hydrolases isolation & purification, N-Glycosyl Hydrolases metabolism, Plant Lectins, Plant Proteins isolation & purification, Plant Proteins metabolism, Protein Biosynthesis, Ribosome Inactivating Proteins isolation & purification, Ribosome Inactivating Proteins metabolism, Ribosome Inactivating Proteins, Type 2, Ricin metabolism, Temperature, Trees, HeLa Cells drug effects, N-Glycosyl Hydrolases toxicity, Plant Proteins toxicity, Ribosome Inactivating Proteins toxicity, Ricin toxicity
- Abstract
Nigrin b, a lectin isolated from the bark of elderberry (Sambucus nigra L.), has structure and enzymatic activity similar to that of ricin and other type 2 ribosome inactivating proteins (RIPs), and yet is much less toxic to cells and animals. In an attempt to explain this difference, we studied (1) the cytotoxicity of both lectins at 18 and 37 degrees C, and in the presence of substances interfering with intracellular routing, and (2) the binding of nigrin b to, and its uptake and degradation by HeLa cells, in parallel with ricin. As compared with the latter, (1) less nigrin b was bound and more was degraded by cells, with a resulting lower concentration remaining inside the cells, and (2) there is evidence for a different intracellular routing followed by the two lectins. These results may explain at least partly the different cytotoxicity and consequently the lower toxicity to mice of nigrin b compared with ricin.
- Published
- 1997
- Full Text
- View/download PDF
31. Toxicity of ribosome-inactivating proteins-containing immunotoxins to a human bladder carcinoma cell line.
- Author
-
Battelli MG, Polito L, Bolognesi A, Lafleur L, Fradet Y, and Stirpe F
- Subjects
- Humans, Protein Biosynthesis, Ribosome Inactivating Proteins, Type 1, Ribosome Inactivating Proteins, Type 2, Saporins, Tumor Cells, Cultured, Urinary Bladder Neoplasms metabolism, Urinary Bladder Neoplasms pathology, Immunotoxins pharmacology, N-Glycosyl Hydrolases, Plant Proteins pharmacology, Ribosomes drug effects, Urinary Bladder Neoplasms therapy
- Abstract
Immunotoxins were prepared by linking the type 1 ribosome-inactivating proteins (RIP) momordin I, pokeweed antiviral protein from seeds (PAP-S) and saporin-S6 to the 48-127 monoclonal antibody (MAb) recognising a glycoprotein (gp54) expressed on all human bladder tumours tested and on human bladder carcinoma cell lines, in particular on the T24 cell line. T24 cells required a 2 hr contact with immunotoxins to ensure binding and endocytosis. A time course of exposure, followed by further incubation without the immunotoxins, showed that maximum inhibition of protein synthesis by T24 cells was reached after 2 hr of contact followed by 3 days without the immunotoxins. Under optimal conditions, 48-127/RIP immunotoxins at nanomolar concentrations inhibited by 50% protein synthesis of target T24 cells. No toxicity was observed if (i) target cells were treated with non-conjugated RIP, (ii) target cells were treated with momordin I- or PAP-S-containing immunotoxins made with an irrelevant antibody and (iii) a non-target cell line was treated with the same 2 RIP conjugated to 48-127 antibody. The in vitro selective toxicity of these immunotoxins encourages further studies in view of a possible use in clinical trials for the local therapy of human bladder carcinomas.
- Published
- 1996
- Full Text
- View/download PDF
32. Excitotoxic increase of xanthine dehydrogenase and xanthine oxidase in the rat olfactory cortex.
- Author
-
Battelli MG, Buonamici L, Abbondanza A, Virgili M, Contestabile A, and Stirpe F
- Subjects
- Animals, Hippocampus drug effects, Hippocampus enzymology, Kainic Acid pharmacology, Male, Rats, Rats, Wistar, Neurotoxins pharmacology, Olfactory Pathways drug effects, Olfactory Pathways enzymology, Xanthine Dehydrogenase metabolism, Xanthine Oxidase metabolism
- Abstract
Excitotoxic lesions induced by systemic injection of kainic acid, resulted in 2-3-fold increase of xanthine dehydrogenase and xanthine oxidase activities in the rat olfactory cortex 48-72 h after drug administration. A significant increase of the xanthine oxidase/dehydrogenase ratio was also observed at 4 and 48 h post-injection. No similar changes were noticed in the hippocampus. The enhancement of enzyme activity seems to be primarily a consequence of the altered cell composition in damaged area. Free radicals produced by the increased oxygen-dependent form of the enzyme could in turn aggravate the excitotoxic brain injury.
- Published
- 1995
- Full Text
- View/download PDF
33. Ribosome-inactivating proteins (RNA N-glycosidases) from the seeds of Saponaria ocymoides and Vaccaria pyramidata.
- Author
-
Bolognesi A, Olivieri F, Battelli MG, Barbieri L, Falasca AI, Parente A, Del Vecchio Blanco F, and Stirpe F
- Subjects
- Amino Acid Sequence, Animals, Cell Line, Humans, Lethal Dose 50, Mice, Molecular Sequence Data, N-Glycosyl Hydrolases isolation & purification, N-Glycosyl Hydrolases metabolism, N-Glycosyl Hydrolases toxicity, Plant Proteins isolation & purification, Plant Proteins toxicity, Plants embryology, Protein Synthesis Inhibitors chemistry, Protein Synthesis Inhibitors isolation & purification, Protein Synthesis Inhibitors pharmacology, Rats, Ribosome Inactivating Proteins, Ribosomes metabolism, Tumor Cells, Cultured, N-Glycosyl Hydrolases pharmacology, Plant Proteins pharmacology, Plants enzymology, Ribosomes drug effects
- Abstract
From the seeds of the Caryophyllaceae Saponaria ocymoides and Vaccaria pyramidata two proteins were purified which have the properties of the type-1 (single-chain) ribosome-inactivating proteins [reviewed by Barbieri, L., Battelli, M. G. & Stirpe, F. (1993) Ribosome-inactivating proteins from plants, Biochim. Biophys. Acta 1154, 237-282]. The proteins have molecular masses of 30.2 kDa (S. ocymoides) and 28.0 kDa (V. pyramidata) and pI greater than 9.5, their N-terminal amino acid sequences are similar to those of saporin-S6 and dianthin 30, ribosome-inactivating proteins from other Caryophyllaceae, and they partially cross-react with sera against these proteins. Both proteins inhibit protein synthesis by a rabbit-reticulocyte lysate with IC50 (concentrations giving 50% inhibition) below 10(-10) M, have a smaller effect on poly(U)-directed phenylalanine polymerisation by rat liver ribosomes (nanomolar IC50, approximately) and on protein synthesis by various cell lines (IC50 ranging from 4 nM to > 3000 nM) and possess rRNA N-glycosidase activity, releasing 1 mol adenine/ribosome.
- Published
- 1995
- Full Text
- View/download PDF
34. In vivo and in vitro uptake of an anti-CD30/saporin immunotoxin by rat liver parenchymal and nonparenchymal cells.
- Author
-
Battelli MG, Buonamici L, Bolognesi A, and Stirpe F
- Subjects
- Animals, Antibodies, Monoclonal pharmacokinetics, Antibodies, Monoclonal pharmacology, Antineoplastic Agents, Phytogenic pharmacology, Cells, Cultured, Female, Immunotoxins pharmacology, Kupffer Cells metabolism, Liver cytology, Plant Proteins pharmacology, Protein Biosynthesis, Rats, Ribosome Inactivating Proteins, Type 1, Saporins, Tissue Distribution, Antineoplastic Agents, Phytogenic pharmacokinetics, Immunotoxins pharmacokinetics, Ki-1 Antigen immunology, Liver metabolism, N-Glycosyl Hydrolases, Plant Proteins pharmacokinetics
- Abstract
A Ber-H2/saporin immunotoxin, consisting of the single-chain ribosome-inactivating protein saporin-S6 and the anti-CD30 monoclonal antibody Ber-H2, gave encouraging results in the treatment of refractory Hodgkin's disease but caused a transient hepatotoxicity. The accumulation of Ber-H2/saporin conjugate and of its components by rat liver parenchymal and nonparenchymal cells was studied. The in vivo concentration of intravenously injected Ber-H2/saporin, saporin or Ber-H2 in nonparenchymal cells was 4-, 25- and 11-fold higher, respectively, than that in parenchymal cells. Adherent in vitro cultured nonparenchymal cells, mostly Kupffer cells, accumulated the proteins approximately 10 times more than parenchymal cells; traces of free saporin were taken up by both types of cells. In vitro protein synthesis by both cell types was inhibited by 50% at nanomolar concentrations of saporin. Nonparenchymal cells were sensitive to Ber-H2/saporin at picomolar concentrations, whereas parenchymal cells were unaffected by the immunotoxin up to 100 pmol/L. The results of the uptake of, and the sensitivity to, the immunotoxin suggest that the sensitivity of liver cells is proportional to the uptake and that the in vivo damage to parenchymal cells is at least in part mediated by the toxicity to nonparenchymal liver cells.
- Published
- 1994
- Full Text
- View/download PDF
35. Ribosome-inactivating proteins from plants.
- Author
-
Barbieri L, Battelli MG, and Stirpe F
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Glycoside Hydrolases biosynthesis, Humans, Molecular Sequence Data, Plant Proteins biosynthesis, Plant Proteins pharmacology, Plant Proteins toxicity, Terminology as Topic, Plant Proteins chemistry
- Published
- 1993
- Full Text
- View/download PDF
36. Purification and partial characterization of single-chain ribosome-inactivating proteins from the seeds of Phytolacca dioica L.
- Author
-
Parente A, De Luca P, Bolognesi A, Barbieri L, Battelli MG, Abbondanza A, Sande MJ, Gigliano GS, Tazzari PL, and Stirpe F
- Subjects
- Amino Acid Sequence, Animals, Cell Line drug effects, Female, Glycoside Hydrolases chemistry, Glycoside Hydrolases toxicity, Mice, Molecular Sequence Data, Plant Proteins chemistry, Plant Proteins toxicity, Protein Biosynthesis, Rabbits, Rats, Ribosome Inactivating Proteins, Type 1, Ribosomes drug effects, Saporins, Glycoside Hydrolases isolation & purification, Immunotoxins isolation & purification, N-Glycosyl Hydrolases, Plant Proteins isolation & purification, Ribosomes metabolism, Seeds chemistry
- Abstract
Three ribosome-inactivating proteins (RIPs) similar to those already known (Stirpe et al. (1992) Bio/Technology 10, 405-412) were purified from the seeds of Phytolacca dioica. These proteins, called Phytolacca dioica RIPs (PD-S1, PD-S2 and PD-S3 RIPs), are glycoproteins, with M(r) approx. 30,000, inhibit protein synthesis by a rabbit reticulocyte lysate and phenylalanine polymerization by isolated ribosomes, and depurinate rat liver rRNA in an apparently identical manner as the A-chain of ricin and other RIPs (Endo et al. (1987) J. Biol. Chem. 262, 5908-5912). Part of the purified rat liver ribosomes appeared resistant to the action of PD-S RIPs. The most abundant protein, PD-S2 RIP, gave a weak or nil cross-reaction with sera against various other RIPs, including a pokeweed antiviral protein from the roots of Phytolacca americana. PD-S2 RIP was linked to a monoclonal antibody (Ber-H2) against the CD30 human lymphocyte antigen and the resulting immunotoxin was selectively toxic to the CD30 + Hodgkin's lymphoma-derived L540 cell line.
- Published
- 1993
- Full Text
- View/download PDF
37. Distribution and properties of major ribosome-inactivating proteins (28 S rRNA N-glycosidases) of the plant Saponaria officinalis L. (Caryophyllaceae).
- Author
-
Ferreras JM, Barbieri L, Girbés T, Battelli MG, Rojo MA, Arias FJ, Rocher MA, Soriano F, Mendéz E, and Stirpe F
- Subjects
- Amino Acid Sequence, Amino Acids analysis, Aniline Compounds, Animals, Cell Line, Cell-Free System drug effects, Escherichia coli, Humans, Molecular Sequence Data, N-Glycosyl Hydrolases chemistry, Plant Proteins chemistry, Plant Proteins pharmacology, Protein Biosynthesis drug effects, Rabbits, Rats, Ribosome Inactivating Proteins, Ribosome Inactivating Proteins, Type 1, Ribosomes drug effects, Saporins, Immunotoxins, N-Glycosyl Hydrolases isolation & purification, Plant Proteins isolation & purification, Ribosomes metabolism
- Abstract
We have studied the distribution of the protein synthesis inhibitory activity in the tissues of Saponaria officinalis L. (Caryophyllaceae). Seven major saporins, ribosome-inactivating proteins, were purified to apparent homogeneity from leaves, roots and seeds using a new procedure of RIPs isolation including ion-exchange and hydrophobic chromatography. They all catalysed the depurination of rat liver ribosomes, which generate the Endo's diagnostic rRNA fragment upon treatment with acid aniline, thus indicating that A4324 from the 28S rRNA has been released (Endo et al. (1987) J. Biol. Chem. 262, 5908-5912). The molecular mass of saporins by SDS-PAGE ranged between 30.2 and 31.6 kDa and by gel-filtration between 27.5 and 30.1 kDa. Amino acid composition and amino-terminal amino acid sequence indicate that all saporins may be considered isoforms. Only two saporins present in roots were glycosylated (SO-R1 and SO-R3). All saporins are very active on cell-free translation systems derived from rabbit reticulocyte lysates, rat liver, Triticum aestivum L., Cucumis sativus L. and Vicia sativa L. However, they are poor inhibitors of an Escherichia coli translation system. They inhibit protein synthesis in HeLa, BeWo and NB 100 cells, HeLa cells being the most resistant. The enzymatic activity of at least one saporin isoform was dependent on magnesium concentration in the standard rat liver cell-free system.
- Published
- 1993
- Full Text
- View/download PDF
38. High sensitivity of cultured human trophoblasts to ribosome-inactivating proteins.
- Author
-
Battelli MG, Montacuti V, and Stirpe F
- Subjects
- Binding Sites, Estradiol metabolism, Humans, Ribosome Inactivating Proteins, Type 1, Ribosome Inactivating Proteins, Type 2, Saporins, Trophoblasts cytology, Trophoblasts metabolism, Tumor Cells, Cultured, Abortifacient Agents pharmacology, Immunotoxins, N-Glycosyl Hydrolases, Plant Proteins pharmacology, Ribosomes drug effects, Trophoblasts drug effects
- Abstract
Many plant proteins possessing abortifacient activities were identified as ribosome-inactivating proteins (RIPs). The effect of several ribosome-inactivating proteins (saporin 6, dianthin 32, pokeweed antiviral protein from seeds, gelonin, bryodin-R, and momordin) on primary cultures of human trophoblasts and human embryonal fibroblasts and on choriocarcinoma (JAR and BeWo) and ovarian carcinoma (TG) cell lines was studied. Protein synthesis of human trophoblasts and BeWo cells was lowered by RIPs more than that of other cells. The trophoblastic receptors for estradiol were not affected by treatment of the cells with momordin. The binding and uptake of saporin 6 and momordin by BeWo and HeLa cells were not correlated to cell toxicity.
- Published
- 1992
- Full Text
- View/download PDF
39. Effects of hypoxia and ethanol on xanthine oxidase of isolated rat hepatocytes: conversion from D to O form and leakage from cells.
- Author
-
Battelli MG, Abbondanza A, and Stirpe F
- Subjects
- Animals, Cell Survival drug effects, Cell Survival physiology, Humans, Liver drug effects, Male, Rats, Rats, Inbred Strains, Xanthine Dehydrogenase blood, Xanthine Oxidase blood, Cell Hypoxia physiology, Ethanol pharmacology, Liver cytology, Liver enzymology, Xanthine Dehydrogenase drug effects, Xanthine Oxidase drug effects
- Abstract
The combined effects of ethanol and hypoxia on the conversion of xanthine dehydrogenase (D form) to xanthine oxidase (O form) and on the leakage of the enzyme from isolated rat hepatocytes was studied. Time-dependent death of cells occurred during incubation in hypoxic conditions. Ethanol (40 mM) had only a moderate effect on viability in aerobiosis, but accelerated the loss of hypoxic cells, which was 96% after 3 h of incubation. In hypoxic conditions, the xanthine oxidase was gradually converted from D into O form. The conversion was complete in 3 h, and was accelerated by 1 mM xanthine or by ethanol, in a concentration-related manner. Hypoxia brought about a progressive leakage of xanthine oxidase from hepatocytes, which was accelerated by ethanol in a concentration-dependent manner. The enzyme found outside hepatocytes was mostly in its O form. The xanthine oxidase of hepatocytes cytosol was converted from D into O form by human plasma or serum. In all cases the conversion could be completely reverted by treatment of the extract with dithiothreitol.
- Published
- 1992
- Full Text
- View/download PDF
40. Ribosome-inactivating proteins from plants: present status and future prospects.
- Author
-
Stirpe F, Barbieri L, Battelli MG, Soria M, and Lappi DA
- Subjects
- Adenine metabolism, Amino Acid Sequence, Base Sequence, Humans, Molecular Sequence Data, Plant Proteins chemistry, Sequence Homology, Nucleic Acid, Forecasting, Plant Proteins pharmacology, Ribosomes
- Abstract
Plant ribosome-inactivating proteins (RIPs) are N-glycosidases which cleave the N-glycosidic bond of adenine in a specific ribosomal RNA sequence. Most commonly RIPs are single-chain proteins (type 1 RIPs), but some (type 2 RIPs) possess a galactose-specific lectin domain that binds to cell surfaces. The latter RIPs are potent toxins, the best known of which is ricin. RIPs have antiviral and abortifacient activities, and, in a widespread application, can also be linked to antibodies or ligands to form immunotoxins or conjugates specifically toxic to a given type of cell.
- Published
- 1992
- Full Text
- View/download PDF
41. Cytotoxicity of, and DNA damage by, active oxygen species produced by xanthine oxidase.
- Author
-
Chiricolo M, Tazzari PL, Abbondanza A, Dinota A, and Battelli MG
- Subjects
- Burkitt Lymphoma genetics, Burkitt Lymphoma pathology, Cell Line, Clone Cells metabolism, DNA, Single-Stranded drug effects, Humans, DNA Damage, DNA, Neoplasm drug effects, Free Radical Scavengers, Oxygen toxicity, Xanthine Oxidase toxicity
- Abstract
Toxicity to Raji cells of the xanthine oxidase/hypoxanthine system is related to the formation of single-strand DNA breaks. DNA damage was proportional to the concentration of xanthine oxidase and to the time of exposure. It was prevented by the absence of hypoxanthine, or by the presence of allopurinol, or both superoxide dismutase and catalase. The release of 51Cr from damaged cells was detectable 12 h after the inhibition of cloning efficiency and the production of DNA breakage. These data suggest that DNA damage induced by the oxygen products precedes the severe lesion to the cellular membrane.
- Published
- 1991
- Full Text
- View/download PDF
42. Purification and properties of new ribosome-inactivating proteins with RNA N-glycosidase activity.
- Author
-
Bolognesi A, Barbieri L, Abbondanza A, Falasca AI, Carnicelli D, Battelli MG, and Stirpe F
- Subjects
- Amino Acid Sequence, Animals, Glycoproteins chemistry, Glycoproteins isolation & purification, Glycoproteins metabolism, Glycoproteins toxicity, Humans, In Vitro Techniques, Isoelectric Point, Mice, Molecular Sequence Data, Molecular Weight, N-Glycosyl Hydrolases chemistry, N-Glycosyl Hydrolases metabolism, N-Glycosyl Hydrolases toxicity, Plant Proteins chemistry, Plant Proteins metabolism, Plant Proteins toxicity, Protein Biosynthesis, Rabbits, Ribosome Inactivating Proteins, Ribosome Inactivating Proteins, Type 1, N-Glycosyl Hydrolases isolation & purification, Plant Proteins isolation & purification, RNA, Ribosomal metabolism, Ribosomes metabolism
- Abstract
Ribosome-inactivating proteins (RIPs) similar to those already known (Stirpe & Barbieri (1986) FEBS Lett. 195, 1-8) were purified from the seeds of Asparagus officinalis (two proteins, asparin 1 and 2), of Citrullus colocynthis (two proteins, colocin 1 and 2), of Lychnis chalcedonica (lychnin) and of Manihot palmata (mapalmin), from the roots of Phytolacca americana (pokeweed antiviral protein from roots, PAP-R) and from the leaves of Bryonia dioica (bryodin-L). The two latter proteins can be considered as isoforms, respectively, of previously purified PAP, from the leaves of P. americana, and of bryodin-R, from the roots of B. dioica. All proteins have an Mr at approx, 30,000, and an alkaline isoelectric point. Bryodin-L, colocins, lychnin and mapalmin are glycoproteins. All RIPs inhibit protein synthesis by a rabbit reticulocyte lysate and phenylalanine polymerization by isolated ribosomes and alter rRNA in a similar manner as the A-chain of ricin and related toxins (Endo et al. (1987) J. Biol. Chem. 262, 5908-5912).
- Published
- 1990
- Full Text
- View/download PDF
43. Bone marrow purging by a xanthine oxidase-antibody conjugate.
- Author
-
Dinota A, Tazzari PL, Abbondanza A, Battelli MG, Gobbi M, and Stirpe F
- Subjects
- Bone Marrow immunology, Cytotoxicity, Immunologic, Evaluation Studies as Topic, Humans, In Vitro Techniques, Multiple Myeloma surgery, Plasma Cells immunology, Plasma Cells pathology, Tumor Cells, Cultured immunology, Tumor Cells, Cultured pathology, Antibodies, Monoclonal administration & dosage, Bone Marrow pathology, Bone Marrow Transplantation methods, Xanthine Oxidase administration & dosage
- Abstract
The selective cytotoxicity of the xanthine oxidase conjugated to an 8A monoclonal antibody recognizing a human plasma cell-associated antigen has been described. The selectivity and the toxicity of the hypoxanthine/conjugated xanthine oxidase system was increased by removing the excess of conjugate and by adding chelated iron. Under these experimental conditions the cytotoxicity of the conjugate exceeded that of free xanthine oxidase by one order of magnitude. The conjugate effectively purged bone marrow from infiltrating neoplastic plasma cells and added target Raji cells, provided blood was removed and bone marrow peroxidases were exhausted. In conditions of purging effectiveness the conjugate had no toxicity to CFU-GM. No toxicity to mice was observed after i.v. injection of xanthine oxidase-antibody conjugate up to 2.9 U/kg body weight. Thus the hypoxanthine/conjugated xanthine oxidase system could be an effective and nontoxic tool for the ex vivo bone marrow purging in multiple myeloma patients for autologous transplantation.
- Published
- 1990
44. Toxicity of, and histological lesions caused by, ribosome-inactivating proteins, their IgG-conjugates, and their homopolymers.
- Author
-
Battelli MG, Barbieri L, and Stirpe F
- Subjects
- Animals, Glycoproteins toxicity, Immunoglobulin G, Kidney drug effects, Lethal Dose 50, Liver drug effects, Mice, Necrosis, Rabbits, Reticulocytes drug effects, Reticulocytes metabolism, Ribosome Inactivating Proteins, Type 1, Saporins, Immunotoxins toxicity, Kidney pathology, Liver pathology, N-Glycosyl Hydrolases, Plant Proteins toxicity, Protein Synthesis Inhibitors toxicity, Ribosomes drug effects, Toxins, Biological
- Abstract
The toxicity of, and the lesions brought about by, several ribosome-inactivating proteins (bryodin, gelonin, momordin, pokeweed antiviral protein from seeds, saporin 6, trichokirin and momorcochin-S), either native, or conjugated to bovine IgG, or polymerized, were studied in the mouse. Severe necrotic liver damage was the main lesion present in animals receiving lethal doses of the proteins. The toxicity of ribosome-inactivating proteins increased after conjugation to IgG or homopolymerization. The toxicity of conjugates to mouse was not predictable from the inhibitory activity on cell-free protein synthesis.
- Published
- 1990
- Full Text
- View/download PDF
45. Reduction of ricin and other plant toxins by thiol:protein disulfide oxidoreductases.
- Author
-
Barbieri L, Battelli MG, and Stirpe F
- Subjects
- Animals, Cattle, Kinetics, Liver enzymology, Protein Biosynthesis drug effects, Rabbits, Reticulocytes metabolism, Ribosome Inactivating Proteins, Type 2, Abrin pharmacology, Lectins pharmacology, Oxidoreductases metabolism, Plant Lectins, Plant Proteins pharmacology, Protein Disulfide Reductase (Glutathione) metabolism, Ricin pharmacology, Toxins, Biological pharmacology
- Published
- 1982
- Full Text
- View/download PDF
46. Enzymic conversion of rat liver xanthine oxidase from dehydrogenase (D form) to oxidase (O form).
- Author
-
Battelli MG
- Subjects
- Animals, Cattle, Chromatography, Gel, Hot Temperature, Milk, Oxidation-Reduction, Protein Denaturation, Rats, Sulfhydryl Compounds metabolism, Liver enzymology, Xanthine Oxidase metabolism
- Published
- 1980
- Full Text
- View/download PDF
47. Effect of growth on the estrogen receptor levels in MCF-7 cells.
- Author
-
Brooks SC, Hansen ER, Saunders DE, Battelli MG, and Shafie SM
- Subjects
- Cell Division, Cell Line, Estradiol metabolism, Female, Humans, Kinetics, Receptors, Estradiol, Receptors, Estrogen isolation & purification, Breast Neoplasms physiopathology, Receptors, Estrogen metabolism
- Abstract
MCF-7 cells have been shown to contain estrogen receptor in several cell fractions following homogenization: nuclei, microsomes, and cytosol. The amount of 17 beta-estradiol-binding capacity found in each cellular compartment depended on the inclusion of detergent in homogenization buffers and on the use of 0.25 M sucrose in the nuclear washes. 17 beta-Estradiol receptor (E2R) associated with nuclei (whole nuclei exchange assay, 0.6 M KCl soluble, and that found on membranes sheared from crude nuclear pellets by centrifugation in 0.25 M sucrose buffer) displayed a dissociation constant (Kd) of 0.77 +/- 0.01 (S.D.) nM (n = 7). KdS of the cytoplasmic (microsomes and soluble) receptors were determined to be 0.33 +/- 0.10 nM (n = 9). Exchangeable ligand on partially purified nuclei assumed its highest level in MCF-7 cells during logarithmic growth in serum-containing media (0.8 pmol/micrograms DNA) but declined after the culture reached confluence (0.2 pmol/micrograms DNA). Seventy-five % of the nuclear E2R declined linearly after feeding MCF-7 cells in logarithmic growth phase an estrogen- and serum-free medium (t1/2 3.5 days). Another class of salt-extractable nuclear receptor (0.2 pmol/micrograms DNA) persisted in postconfluent cultures whether fed estrogen (serum-containing media) or not (serum-free media). This residual binding capacity remained in nuclei of MCF-7 cells for an extended period of time. MCF-7 cells demonstrated functionality of E2R throughout their growth phases as evidenced by the replenishment of cytosolic E2R and the induction of progesterone receptor when given 17 beta-estradiol.
- Published
- 1984
48. On the distribution of ribosome-inactivating proteins amongst plants.
- Author
-
Gasperi-Campani A, Barbieri L, Battelli MG, and Stirpe F
- Subjects
- Animals, Chemical Phenomena, Chemistry, Electrophoresis, Polyacrylamide Gel, In Vitro Techniques, Plant Extracts pharmacology, Prokaryotic Initiation Factor-3, Protein Biosynthesis, Rabbits, Reticulocytes metabolism, Peptide Initiation Factors analysis, Plant Proteins pharmacology, Plants analysis, Ribosomes drug effects
- Abstract
The extracts from various parts (mostly seeds) of 56 different plants were examined for inhibition of protein synthesis by a rabbit reticulocyte lysate. Most extracts inhibited protein synthesis with an ID50 (concentration giving 50% inhibition) of 100 micrograms extract protein per ml, or less. The extracts with high activity were partially purified by CM cellulose chromatography. Protein-containing fractions were separated which inhibited protein synthesis and resembled the ribosome-inactivating proteins from plants previously described. Thus, ribosome-inactivating proteins appear to be virtually ubiquitous in plants.
- Published
- 1985
- Full Text
- View/download PDF
49. DNA repair after gamma radiation and superoxide dismutase activity in lymphocytes from subjects of far advanced age.
- Author
-
Licastro F, Franceschi C, Chiricolo M, Battelli MG, Tabacchi P, Cenci M, Barboni F, and Pallenzona D
- Subjects
- Adolescent, Adult, Age Factors, Aged, Gamma Rays, Humans, T-Lymphocytes metabolism, Thymidine metabolism, DNA Repair radiation effects, Lymphocytes enzymology, Superoxide Dismutase blood
- Abstract
DNA repair after gamma radiation was studied in purified T lymphocytes from young and aged subjects. Two different assays were employed. In the first, T lymphocytes were stimulated with phytohemagglutinin (PHA) for 72 h and then treated with hydroxyurea, irradiated with 30 K rads and pulsed with [3H]thymidine (TdR) for 4 h. In the second, T lymphocytes were first irradiated with graded doses of gamma rays (200-800 rads) and then stimulated with PHA, cultured for 72 h and pulsed with 3H-TdR for the last 6 h of culture. T lymphocytes from aged subjects showed a lack of DNA repair synthesis in the first assay whereas only minor differences were found in the second assay between the two groups, i.e., a certain degree of radioresistance in aged lymphocytes. Lymphocyte superoxide dismutase activity showed great individual variations in both groups and a slight increase in old subjects.
- Published
- 1982
- Full Text
- View/download PDF
50. Effect of ribosome-inactivating proteins on ribosomes from Tetrahymena pyriformis and Acanthamoeba castellanii.
- Author
-
Cenini P, Battelli MG, Bolognesi A, Stirpe F, and Villemez CL
- Subjects
- Acanthamoeba drug effects, Animals, Kinetics, Magnesium pharmacology, Peptide Biosynthesis, Protein Biosynthesis drug effects, Ribosomes drug effects, Tetrahymena pyriformis drug effects, Acanthamoeba genetics, Peptides, Ribosomes metabolism, Tetrahymena pyriformis genetics, Toxins, Biological pharmacology
- Abstract
The effect of ribosome-inactivating proteins type 1 (single-chain) and type 2 (two-chain, toxins) on polyphenylalanine polymerization by Tetrahymena pyriformis and Acanthamoeba castellanii ribosomes has been studied. The reaction catalysed by tetrahymena ribosomes was inhibited by two ribosome-inactivating proteins type 1 (dianthin 32 and, less effectively, momordin) whereas the reaction catalysed by amoeba ribosomes was inhibited, in a decreasing order of activity, by three ribosome-inactivating proteins type 1 (dianthin 32, saporin 6 and bryodin) and by two toxins (abrin and volkensin).
- Published
- 1987
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.