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G Poli

Publications and source records attributed to G Poli.

At least 145 records · Page 8Linked to original sources
PubMed1996

Interleukin-10-induced HIV-1 expression is mediated by induction of both membrane-bound tumour necrosis factor (TNF)-alpha and TNF receptor type 1 in a promonocytic cell line.

OBJECTIVE: To investigate whether the upregulatory effect of interleukin (IL)-10 on HIV expression in a model of latent HIV infection is mediated by induction of endogenous tumour necrosis factor (TNF)-alpha and TNF receptors (TNFR). DESIGN: The latently HIV-infected promonocytic cell line U1 was examined, because in this in vitro model IL-10 has been shown to synergize with multiple cytokines, including TNF-alpha, in enhancing HIV production. METHODS: Membrane-bound TNF-alpha, TNFR-1 and TNFR-2 surface expression were determined by flow cytometry. TNF-alpha mRNA was estimated by competitive polymerase chain reaction (PCR), and TNF-alpha, soluble TNFR-1 and soluble TNFR-2 supernatant content by enzyme-linked immunosorbent assay. HIV-1 expression was quantitated by reverse transcriptase assay and p24 antigen release. RESULTS: We demonstrated that IL-10 induces a time and cell-concentration dependent upregulation of HIV expression in U1 cells. This effect is mediated through the endogenous production of TNF-alpha as demonstrated by blocking experiments with anti-TNF-alpha antibodies and by detection of IL-10-induced increase of TNF-alpha mRNA by competitive PCR. More importantly, IL-10 is able to upregulate membrane-bound TNF-alpha and TNFR-1, along with a consistent increase in the shedding of soluble TNFR-1 without inducing detectable TNF-alpha secretion. CONCLUSIONS: IL-10 activates HIV expression through the membrane-bound TNF-alpha/TNFR-1 pathway, suggesting an amplification mechanism of HIV expression that might occur during cell-to-cell interaction. This positive regulatory effect of IL-10 in an in vitro model of chronic HIV infection is consistent with the inexorable progression of disease seen in advanced patients when both IL-10 and TNF-alpha are elevated.

Antigens, CD↗
PubMed1996

Molecular characterization of a variant of proviral bovine leukaemia virus (BLV).

Southern-blot hybridization and partial sequencing of the pol and env genes were used to characterize BLV-integrated provirus of seropositive cattle from two dairy herds in northern Italy. Comparison of the data obtained with those of previously characterized BLV strains from other geographic areas (Australia, Belgium, Japan and USA) revealed the presence of a viral variant (BLV-12), which showed both conserved and unique features. Regarding the gp51 envelope glycoprotein, the BLV-12 variant showed: 1. A high extent of conservation, which included potential glycosylation sites and cysteine residues; 2. Three unique amino acid residues not present in any of the other BLV strains analysed; and 3. Some variability at the level of one (G) of the three (F, G and H) conformational epitopes, which is probably important in the process of infection. These results agree with the suggestion that the sequence variability of the gp51 glycoprotein preferentially involves structures whose change is thought to underlie the phenomenon of escape from immune surveillance.

Amino Acid Sequence↗
PubMed1996

Surface analysis of IgM-expressing bovine lymphocytes in BLV-infected cattle by immuno-scanning electron microscopy.

Surface of IgM-expressing lymphocytes infected by bovine leukemia virus and with/without persistent lymphocytosis (BLV+PL-, BLV+PL+) in cattle is heterogenous. Three distinct morphological structures of B lymphocytes were found in these groups by immuno-scanning electron microscopy (ISEM). B lymphocytes with generally elongated and pleomorphic microvilli and sometimes short ruffles were observed in infected, and as well as in non-infected animals. A second morphological type endowed with a relatively small number of elongated villi-like veils, and a third morphological type characterized by a relatively small number of short microvilli or blunt stub-like microvilli covering the smooth surface were seen, mainly in BLV+PL+ and seldom in BLV+PL- Cells of the second type were more intensively labeled, than lymphocytes of the third type.

Animals↗
PubMed1996

Association of BoLA class II haplotypes with subclinical progression of bovine leukaemia virus infection in Holstein-Friesian cattle.

The influence of bovine lymphocyte antigen (BoLA) complex polymorphism on subclinical progression of bovine leukaemia virus (BLV) infection was investigated in 41 Holstein-Friesian cows from two herds in Italy. All cows were seropositive for BLV and 22 had persistent lymphocytosis (PL). BoLA-A specificities were defined by serology, and class II haplotypes were defined based on restriction fragment length polymorphism (RFLP) and polymerase chain reaction (PCR)-RFLP analysis of DQ and DR genes. Chi-square analysis revealed a significant and absolute association of haplotype DQA*3A-DQB*3A-DRB2*2A-DRB3.2*11 with resistance to PL (P chi 2 = 0.028, relative risk (RR) = 0.061). Consistent with this observation, multiple regression analysis revealed that animals carrying this haplotype had lower lymphocyte counts (P = 0.0057). By contrast, haplotype DQA*12-DQB*12-DRB2*3A-DRB3.2*8 was associated with susceptibility to PL (P chi 2 = 0.043, RR = 9.625) and increased lymphocyte counts (P = 0.0537). These results confirm the association of haplotype DQA*3A-DQB*3A-DRB2*2A-DRB3.2*11 with resistance to PL, and substantiate earlier findings of haplotype DQA*12-DQB*12-DRB2*3A-DRB3.2*8 as a risk factor for subclinical progression to PL in BLV-infected Holstein-Friesian cattle.

Animals↗
PubMed1995

Scavenging effect of silipide, a new silybin-phospholipid complex, on ethanol-derived free radicals.

Ethanol metabolism by cytochrome P4502E1 (CYP2E1) produces free radical intermediates, identified as hydroxyethyl radicals. We have observed that in vitro addition or in vivo pretreatment of rats with Silipide, a new 1:1 complex of silybin with phosphatidyl-choline, is able to decrease the spin trapping of hydroxyethyl radicals in microsomes from chronic alcohol-fed rats. This effect is not due to an interference with the metabolism of ethanol by CYP2E1, but is rather related to the capacity of the silybin molecule to scavenge hydroxyethyl radicals. However, such an effect is lost when pure silybin in amounts comparable to those present in Silipide is administered instead, due to the low bioavailability of uncomplexed flavonoid. Further experiments in vivo have shown that Silipide administration also decreases hydroxyethyl radical signals detectable in the bile of rats acutely treated with ethanol. The ability of Silipide to scavenge ethanol-derived radicals along with its antioxidant activity suggests that this drug might be potentially useful in counteracting free radical-mediated injuries involved in the development of liver damage caused by alcohol abuse.

Animals↗
PubMed1995

Hepatocellular metabolism of 4-hydroxy-2,3-nonenal is impaired in conditions of chronic cholestasis.

4-Hydroxy-2,3-nonenal is a major aldehydic end-product of lipid peroxidation known to exert several biological and cytotoxic effects and to be produced during conditions of chronic cholestasis. Here we report that viable hepatocytes isolated from cholestatic livers of bile duct-ligated rats (BDL hepatocytes) show a significantly lower rate of HNE metabolism than control cells. This feature is likely to be the consequence of a significant inhibition in the activity of HNE-metabolizing cytosolic glutathione-S-transferase and alcohol dehydrogenase in BDL hepatocytes. Particulate NADP-dependent aldehyde dehydrogenase was also inhibited. No significant change was found for aldehyde reductase activity. A decreased hepatocellular metabolism of HNE can expose liver parenchymal and non-parenchymal cells to cytotoxic as well as pro-inflammatory and pro-fibrogenic effects of HNE, contributing to the development of chronic cholestatic liver damage.

Alcohol Dehydrogenase↗
PubMed1995

IL-10 synergizes with multiple cytokines in enhancing HIV production in cells of monocytic lineage.

Several cytokines, whose expression is increased in human immunodeficiency virus (HIV)-infected individuals, can enhance virus replication in CD4+ T lymphocytes and mononuclear phagocytes (MP). We have previously reported that interleukin (IL)-10 inhibited HIV replication in acutely infected monocyte-derived macrophages (MDM) at concentrations that completely blocked the production of endogenous tumor necrosis factor-alpha (TNF-alpha) and IL-6 from infected cells. In the present study, lower concentrations of IL-10, which were unable to completely suppress endogenous cytokines, paradoxically enhanced HIV replication in MDM induced by other cytokines. This synergistic induction of HIV expression by IL-10 in combination with TNF-alpha, IL-6, and other cytokines was also observed in the chronically infected promonocytic cell line, U1. The enhancing effect of IL-10 was correlated with an increase in HIV mRNA accumulation and potentiation of phorbol ester-induced long terminal repeat-driven transcription that was independent of the NF-kappa B and Sp1 transcription factors. Thus, IL-10 is a cytokine capable of exerting complex regulatory effects on HIV expression in MP as a function of its own concentration and of the presence of other HIV regulatory cytokines.

Blotting, Northern↗
PubMed1995

Cloning and characterization of a new isoform of the interleukin 1 receptor antagonist.

By reverse transcriptase polymerase chain reaction on messenger RNA from human polymorphonuclear cells, we have isolated a sequence identical to the cDNA coding for intracellular interleukin 1 receptor antagonist (icIL-1ra), but containing an additional in-frame 63-bp sequence located three codons downstream of the translation start of icIL-1ra. This additional sequence is inserted between the first and second exon of the intracellular form, the latter of which is colinear with part of the first exon of the secreted form of IL-1ra. The additional sequence is coded by an extra exon located 2 kb downstream the first icIL-1ra-specific exon. The complementary DNA sequence of the alternatively spliced form of icIL-1ra shows that the predicted protein differs from classical icIL-1ra in the NH2 terminus by insertion of a leaderless sequence of 21 amino acids rich in glycine and glutamic acid residues. Transcripts coding for this new form of icIL-1ra were detected in activated fibroblasts, keratinocytes, and at low levels in myelomonocytic cells. The recombinant protein expressed in COS cells had an apparent molecular mass in sodium dodecyl sulfate polyacrylamide gel electrophoresis of 25 kD compared to 22 kD of classical icIL-1ra, and was mostly intracellular. The ability of this new form of icIL-1ra to inhibit IL-1 activity, in terms of induction of E-selectin and human immunodeficiency virus replication, was comparable to that of classical icIL-1ra. We propose to refer to this new form of icIL-1ra as icIL-1ra type II.

Alternative Splicing↗
PubMed1995

HIV replication in IL-2-stimulated peripheral blood mononuclear cells is driven in an autocrine/paracrine manner by endogenous cytokines.

Replication of HIV is regulated by virus-encoded regulatory proteins, as well as by a variety of cellular factors including cytokines. In the present study, we have investigated the autocrine/paracrine effects of endogenous cytokines on HIV replication in primary PBMCs of healthy HIV seronegative individuals. Addition of rIL-2 to cultures between 0 and 72 h after isolation of PBMCs allowed the replication of primary HIV isolates and laboratory-adapted HIV strains to levels comparable with or greater than those obtained in parallel cultures of autologous PHA-blasts. In this regard, both major cellular targets of HIV infection, CD4+ T lymphocytes and mononuclear phagocytes, were maintained for several weeks in IL-2-stimulated PBMC cultures and virion production was observed in both cell lineages. The kinetics of secretion of several cytokines (such as TNF-alpha, IL-1 beta, IL-6, and IFN-gamma), as well as expression of cellular activation markers, paralleled HIV replication in IL-2-stimulated PBMCs. Endogenous pro-inflammatory cytokines and IFN-gamma played a major role in the regulation of HIV replication in IL-2-stimulated PBMCs, as determined by the ability of several anti-cytokine Abs or antagonists to suppress HIV production; this was not the case in parallel cultures of autologous PHA-blasts. Thus, IL-2-stimulated PBMCs may represent a more physiologic in vitro system than PHA-blasts for the study of HIV infection and replication, and should prove useful in investigating the role of cytokines and other host factors in the regulation of HIV production.

Antibodies, Monoclonal↗
PubMed1995

Metabolism of 4-hydroxy-2-nonenal and aging.

Hepatocytes isolated from male Wistar rats of 2-3 and 20-24 months of age were compared as regards concentration and metabolism of 4-hydroxy-2-nonenal, one of the major aldehyde products of n-6 fatty acid oxidative breakdown. A significant accumulation of fluorescent 4-hydroxy-2-nonenal-membrane lipid adducts was found in the cells from the old rats. The mechanism mainly responsible for such aldehyde accumulation was shown to be the impairment of its enzymatic metabolism. In fact, while endogenous, that is non-stimulated, aldehyde production was not impaired, the reductive pathway of 4-hydroxy-2-nonenal metabolism in hepatocytes from old rats was strongly depressed. The decrease of 4-hydroxy-2-nonenal consumption with age was confirmed in homogenates from liver and kidney, while there were no differences between young and old animals in heart, lung or brain.

Aging↗
PubMed1995

Cytokines in the acquired immunodeficiency syndrome and other infectious diseases.

The pathogenesis of several infectious diseases is frequently related not only to their etiological agents of viral, bacterial, or parasitic nature, but also to the host immune response. Both inflammatory responses and specific immune responses to the invading microorganisms are controlled by complex networks of intercellular signalling molecules, namely cytokines. This rapidly growing family of mediators includes lymphokines, interleukins, and molecules such as tumor necrosis factors and interferons. Patterns of cytokine production from antigen- or allergen-specific T lymphocytic cell clones have been identified, firstly in animal models and subsequently in man, and are commonly referred to as TH0, TH1, and TH2 profiles. The predominance of one of these profiles strongly influences the type of immune response (humoral versus cellular) and, at least in some experimental models, whether the immune response is protective or harmful. This is most convincingly demonstrated in models of parasitic diseases, but has also been hypothesized to be involved in the pathogenesis of human immunodeficiency virus infection and the acquired immunodeficiency syndrome. Earlier studies on human immunodeficiency virus infection revealed that the replicative capacity of this retrovirus, like the herpes viruses, is strongly influenced by the cytokine milieu of infected cells. Transcriptional and post-transcriptional regulation of human immunodeficiency virus expression by human cytokines are examples of the complex interdigitation of viruses with the host immune system.

Acquired Immunodeficiency Syndrome↗
PubMed1995

Cytolysis does not per se induce lipid peroxidation: evidence in man.

An increasing bulk of data counters the opinion that cell death and lysis necessarily trigger the formation and release of detectable amounts of molecules that are markers of lipid peroxidation. Plasma levels of thiobarbituric-acid-reacting compounds, protein-aldehyde fluorescent adducts, lipid peroxides, and endogenous antioxidant compounds were monitored versus controls, during intensive care treatment, in six patients seriously poisoned by ingestion of the mushroom Amanita Phalloides. All six patients showed cytolysis, and four of them massive tissue necrosis, as monitored in terms of serum transaminases. In all six patients, however, the blood parameters of redox equilibrium measured were within the normal range for the whole observation period.

Adult↗
PubMed1995

The "bystander effect": association of U-87 cell death with ganciclovir-mediated apoptosis of nearby cells and lack of effect in athymic mice.

Cells expressing the herpes simplex-thymidine kinase (HS-TK) gene as a consequence of retroviral transduction, as well as TK-negative (TK-) bystander cells, can be killed by treatment with ganciclovir (GCV). In vitro, this "bystander effect," has been attributed to metabolic cooperation through gap junctions or to the uptake of apoptotic vesicles. We show that GCV treatment kills TK-negative U-87 glioma cells cocultured with cells that express TK (TK+) but that have lost the capacity for releasing retroviral particles. A photometric enzyme immunoassay identifies histone-associated DNA fragments, typical of apoptosis, in the cytosol of GCV-treated TK+ cells, and apoptotic features are also demonstrated by ultrastructural studies. Northern blot analysis and the reverse transcription polymerase chain reaction (PCR) show that connexin 43, a major constituent of gap junctions, is expressed in TK+ and U-87 cells. The size of U-87 tumors in nude mice subsequently injected with TK+ cells and GCV is not significantly different than in untreated animals; whereas, after injecting 1:1 mixtures of U-87 and TK+ cells, GCV treatment only causes a temporary regression of tumor growth. On the contrary, when the injected mixtures contain PA317.STK.SBA (a retroviral producer cell line that can transduce efficiently the HS-TK gene) and U-87 cells, tumors are destroyed effectively by GCV treatment. Thus, an experimental setting in which U-87 gliomas are matched with cells that are able to express, but not to transduce, the HS-TK gene indicates that the bystander effect kills U-87 cells in vitro by mechanisms associated with apoptotic death. In vivo, this effect is not sufficient to restrain the tumor growth taking place in immunodeficient animals.

Animals↗
PubMed1995

Phylogeny inferred from codon usage pattern in 31 organisms.

We employed the Codonusage database to analyze the codon usage pattern in 31 organisms from all the main biological taxa. We tested the similarity in codon usage pattern between each organism and all the others by the Pearson linear correlation coefficient. The 465 values obtained were located in a 31 x 31 triangular matrix from which a correlation distance matrix was calculated. An evolutionary dendrogram was then constructed from these distances. The results showed a fair correlation between codon usage patterns and phylogenetic relationships, at least for organisms which diverged in rather recent times (end of Jurassic--beginning of Cretaceous).

Animals↗
PubMed1995

Effect of cellular differentiation on cytokine-induced expression of human immunodeficiency virus in chronically infected promonocytic cells: dissociation of cellular differentiation and viral expression.

Cellular differentiation is thought to play an important role in the susceptibility of monocytic lineage cells to human immunodeficiency virus (HIV) infection as well as in their ability to support virus replication. In addition, virus replication in monocytes/macrophages has been demonstrated in vitro to be strongly modulated by several cytokines such as tumor necrosis factor alpha and granulocyte-macrophage colony-stimulating factor. The purpose of the present study was to investigate the interaction between cellular differentiation and cytokines in the regulation of HIV expression from chronically infected monocytic lineage cells. U1, a persistently HIV-infected promonocytic cell line, is characterized by low levels of virus expression which can be modulated by several cytokines. 1 alpha,-25-Dihydroxyvitamin D3 (Vit.D3), a well-known differentiating agent for myelomonocytic cells which has been previously reported to modulate HIV replication in other in vitro systems, induced maturation of U1 cells toward a macrophage-like phenotype, as demonstrated by the induction of the differentiation-associated cell surface markers CD14 and CD11b. Vit.D3-induced differentiation did not result in induction of HIV expression; however, when U1 cells were stimulated with tumor necrosis factor alpha in the presence of Vit.D3, a synergistic induction of cell differentiation and viral expression was demonstrated. In contrast, Vit.D3 suppressed the induction of HIV expression in U1 cells stimulated with gamma interferon, interleukin-6, and granulocyte-macrophage colony-stimulating factor, although synergy between Vit.D3 and these cytokines was observed in terms of cellular differentiation. These data suggest that differentiation of monocytic cells does not necessarily correlate with increased HIV expression.

Calcitriol↗
PubMed1995

Ethanol-induced potentiation of rat hepatocyte damage due to 1,2-dibromoethane.

Low amounts of 1,2-dibromoethane (DBE), not able per se to exert pro-oxidant and cytotoxic activity on rat hepatocyte suspensions, become effective when administered with carbon tetrachloride (CCl4), due to impairment of the glutathione transferase detoxication pathway by CCl4. Treatment of rats with a single dose of ethanol (2.5 g/kg body wt) 2 h before liver cell isolation potentiates the effect of DBE alone on both malonaldehyde formation and lactate dehydrogenase release by the hepatocyte. The potentiation of the DBE effects by ethanol may be through a series of mechanisms, such as a strong inactivation of hepatocyte glutathione transferase similar to that caused by CCl4, an increased basal level of lipid peroxidation and a significant loss of total glutathione.

Animals↗