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Chlamydia pneumoniae replicates in Kupffer cells in mouse model of liver infection

World Journal of Gastroenterology 2006年第40期12卷 6453-6457页

作者： Antonella Marangoni Manuela Donati Francesca Cavrini Rita Aldini Silvia Accardo Vittorio Sambri marco montagnani Roberto Cevenini Sezione di Microbiologia DMCSS University of Bologna Bologna Italy Dipartimento SMETC University of Bologna Bologna Italy Dipartimento di Medicina Interna e Gastroenterologia University of Bologna Bologna Italy

AIM: To develop an animal model of liver infection with Chlamydia pneumoniae (C. pneumoniae) in intraperito-neally infected mice for studying the presence of chlamy-diae in Kupffer cells and ***: A total of 80 BALB/c mice were inoculated intraperitoneally with C. pneumoniae and sacrificed at various time points after infection. Chlamydiae were looked for in liver homogenates as well as in Kupffer cells and hepatocytes separated by liver perfusion with collagenase. C. pneumoniae was detected by both isola-tion in LLC-MK2 cells and fluorescence in situ hybridiza-tion (FISH). The releasing of TNFA-α by C. pneumoniae in vitro stimulated Kupffer cells was studied by enzyme-linked immunosorbent ***: C. pneumoniae isolation from liver homoge-nates reached a plateau on d 7 after infection when 6 of 10 animals were positive, then decreased, and became negative by d 20. C. pneumoniae isolation from sepa-rated Kupffer cells reached a plateau on d 7 when 5 of 10 animals were positive, and became negative by d 20. The detection of C. pneumoniae in separated Kupffer cells by FISH, confirmed the results obtained by culture. Isolated hepatocytes were always negative. Stimula-tion of Kupffer cells by alive C. pneumoniae elicited high TNF-α levels. CONCLUSION: A productive infection by C. pneumo-niae may take place in Kupffer cells and C. pneumoniae induces a local pro-inflammatory activity. C. pneumoniae is therefore, able to act as antigenic stimulus when local-ized in the liver. One could speculate that C. pneumoniaeinfection, involving cells of the innate immunity such as Kupffer cells, could also trigger pathological immune re-actions involving the liver, as observed in human patients with primary biliary cirrhosis.

关键词： Chlamydia pneumoniae Liver infection Kupffer cells Hepatocytes Culture-isolation Fluorescence in situ hybridization TNF-α Primary biliary cirrhosis

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Analysis of ileal sodium/bile acid cotransporter and related nuclear receptor genes in a family with multiple cases of idiopathic bile acid malabsorption

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World Journal of Gastroenterology 2006年第47期12卷 7710-7714页

作者： marco montagnani Anna Abrahamsson Cecilia Glman Gsta Eggertsen Hanns-Ulrich Marschall Elisa Ravaioli Curt Einarsson Paul A Dawson Dipartimento di Medicina Interna e Gastroenterologia Università di Bologna Bologna Italy Center for Applied Biomedi-cal Research Università di Bologna Bologna Italy Center for Gastroenterology and Hepatology Karolinska University Hospital Huddinge Stockholm Sweden Center for Metabolism and Endocrinology Karolinska University Hospital Huddinge Stockholm Sweden Department of Laboratory Medicine Division of Clinical Chemistry Karolinska University Hospital Huddinge Stockholm Sweden Department of Internal Medicine Wake Forest University School of Medicine Winston-Salem NC 27157 United States

The etiology of most cases of idiopathic bile acid malabsorption （IBAH） is unknown. In this study, a Swedish family with bile acid malabsorption in three consecutive generations was screened for mutations in the ileal apical sodium-bile acid cotransporter gene （ASBT; gene symbol, SLC10A2） and in the genes for several of the nuclear receptors known to be important for ASBT expression： the farnesoid X receptor （FXR） and peroxisome proliferator activated receptor alpha （PPARα）. The patients presented with a clinical history of idiopathic chronic watery diarrhea, which was responsive to cholestyramine treatment and consistent with IBAH. Bile acid absorption was determined using ^75Se-homocholic acid taurine （SeHCAT）; bile acid synthesis was estimated by measuring the plasma levels of 7α-hydroxy-4-cholesten-3-one （C4）. The ASBT, FXR, and PPARα genes in the affected and unaffected family members were analyzed using single stranded conformation polymorphism （SSCP）, denaturing HPLC, and direct sequencing. No ASBT mutations were identified and the ASBT gene did not segregate with the bile acid malabsorption phenotype. Similarly, no mutations or polymorphisms were identified in the FXR or PPARα genes associated with the bile acid malabsorption phenotype. These studies indicate that the intestinal bile acid malabsorption in these patients cannot be attributed to defects in ASBT. In the absence of apparent ileal disease, alternative explanations such as accelerated transit through the small intestine may be responsible for the IBAM.

关键词： Bile acid malabsorption Diarrhea Genetics ^75Se-homocholic acid taurine test Nuclear receptors

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Production of reactive oxygen species and expression of inducible nitric oxide synthase in rat isolated Kupffer cells stimulated by Leptospira interrogans and Borrelia burgdorfen

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World Journal of Gastroenterology 2006年第19期12卷 3077-3081页

作者： Antonella Marangoni Silvia Accardo Rita Aldini Massimo Guardigli Francesca Cavrini Vittorio Sambri marco montagnani Aldo Roda Roberto Cevenini Sezione di Microbiologia DMCSS University of Bologna BolognaItaly Istituto di Scienze Chimiche University of Bologna BolognaItaly Dipartimento di Scienze Farmaceutiche University of Bologna BolognaItaly Dipartimento di Medicina Interna e Gastroenterologia University of Bologna BolognaItaly

AIM： To evaluate the production of reactive oxygen species （ROS） and the expression of inducible nitric oxide synthase （iNOS） in rat isolated Kupffer cells （KCs） stimulated by Leptospira interrogans and Borrelia burgdorferi. METHODS： Rat Kupffer cells were separated by perfusion of the liver with 0.05% collagenase, and purified by Percoll gradients. Pudfied Kupffer cells were tested in vitro with alive *** and B. burgdorferi preparations. The production of ROS was determined by chemiluminescence, whereas iNOS protein expression was evaluated by Western blot assay using anti-iNOS antibodies. RESULTS： B. burgdorferi and to a less extent L. interrogans induced ROS production with a peak 35 min after infection. The chemiluminescence signal progressively diminished and was undetectable by 180 min of incubation. Leptospirae and borreliae induced an increased iNOS expression in Kupffer cells that peaked at 6 hours and was still evident 22 h after infection. CONCLUSION： Both genera of spirochetes induced ROS and iNOS production in rat Kupffer cells. Since the cause of liver damage both in leptospiral as well as in borrelial infections are still unknown, we suggest that leptospira and borrelia damage of the liver can be initially mediated by oxygen radicals, and is then maintained at least in part by nitric oxide.

关键词： Leptospira interrogans Borrelia burgdorferi Inducible nitric oxide synthase Nitric oxide Reactive oxygen species Kupffer cells Chemiluminescence

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Severe immune thrombocytopenia after peg-interferonalpha2a, ribavirin and telaprevir treatment completion: a case report and systematic review of literature

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World Journal of Hepatology 2015年第12期7卷 1718-1722页

作者： Rosario Arena Paolo Cecinato Andrea Lisotti Federica Buonfiglioli Claudio Calvanese Giuseppe Grande marco montagnani Francesco Azzaroli Giuseppe Mazzella Department of Medical and Surgical Science-DIMEC University of Bologna S.Orsola-Malpighi Hospital

Mild to moderate autoimmune thrombocytopenia(AITP) is a common finding in patients receiving interferonbased antiviral treatment, due to bone marrow suppression. Here we report the case of a patient with chronic genotype 1b hepatitis C virus(HCV) infection treated with pegylated-interferon alpha-2a, ribavirin and telaprevir for 24 wk; the patient developed severe AITP three weeks after treatment withdrawal. We performed a systematic literature search in order to review all published cases of AITP related to HCV antiviral treatment. To our knowledge, this is the second case of AITP observed after antiviral treatment withdrawal. In most published cases AITP occurred during treatment; in fact, among 24 cases of AITP related to interferonbased antiviral treatment, only one occurred after discontinuation. Early diagnosis of AITP is a key factor in order to achieve an early interferon discontinuation; in the era of new direct antiviral agents those patients have to be considered for interferon-free treatment regimens. Prompt prescription of immuno-suppressant treatment(i.e., corticosteroids, immunoglobulin infusion and even rituximab for unresponsive cases) leads to favourable prognosis in most of cases. Physicians using interferonbased treatments should be aware that AITP can occur both during and after treatment, specially in the new era of interferon-free antiviral treatment. Finally, in the case of suspected AITP, presence of anti-platelet antibodies should be checked not only during treatment but alsoafter discontinuation.

关键词： Autoimmune thrombocytopenia Pegylatedinterferon Chronic hepatitis C Viral hepatitis Antiplateletantibody

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Gastric autoimmune disorders in patients with chronic hepatitis C before,during and after interferon-alpha therapy

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World Journal of Gastroenterology 2003年第7期9卷 1487-1490页

作者： Carlo Fabbri M.Francesca Jaboli Silvia Giovanelli Francesco Azzaroli Alessandro Pezzoli Esterita Accogli Stefania Liva Giovanni Nigro Anna Miracolo Davide Festi Antonio Colecchia marco montagnani Enrico Roda Giuseppe Mazzella Department of Internal Medicine and Gastroenterology University of BolognaBolognaItaly Department of Medicine and Aging University"G D'Annunzio"ChietiItaly

AIM:To explore the prevalence of autoimmune gastritis in chronic hepatitis C virus (HCV) patients and the influence of α-interferon (IFN) treatment on autoimmune gastritis. METHODS:We performed a prospective study on 189 patients with positive anti-HCV and viral RNA enrolled in a 12-month IFN *** evaluated:a) the baseline prevalence of autoimmune gastritis,b) the impact of IFN treatment on development of biochemical signs of autoimmune gastritis (at 3,6 and 12 months),c) the evolution after IFN withdrawal (12 months) in terms of anti-gastric-parietal-cell antibodies (APCA),gastrin,anti-thyroid,and anti-non-organ- specific antibodies. RESULTS:APCA positivity and 3-fold gastrin levels were detected in 3 (1.6%) and 9 (5%) patients,respectively,at baseline,in 25 (13%) and 31 (16%) patients at the end of treatment (both P<0.001,vs baseline),and in 7 (4%) and 14 (7%) patients 12 months after withdrawal (P=0.002 and P=0.01 respectively,vs baseline;P=not significant vs end of treatment).The development of autoimmune gastritis was strictly associated with the presence of autoimmune thyroiditis (P=0.0001),no relationship was found with other markers of autoimmunity. CONCLUSION:In HCV patients,IFN frequently precipitates latent autoimmune gastritis,particularly in *** our 12-month protocol,the phenomenon generally *** APCA positivity and high gastrin levels are associated with the presence of antithyroid antibodies, development of autoimmune thyroiditis during IFN treatment may provide a surrogate preliminary indicator of possible autoimmune gastritis to limit the need for invasive examinations.

关键词： Adult Aged Antiviral Agents dosage Autoimmune Diseases Female Gastrins Gastritis Helicobacter Infections Helicobacter pylori Hepatitis C, Chronic Humans Interferon-alpha Male Middle Aged Organ Specificity Parietal Cells, Gastric Prevalence Prospective Studies Thyroid Gland Treatment Outcome

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Interferon plus ribavirin and interferon alone in preventing hepatocellular carcinoma: A prospective study on patients with HCV related cirrhosis

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World Journal of Gastroenterology 2004年第21期10卷 3099-3102页

作者： Francesco Azzaroli Esterita Accogli Giovanni Nigro Davide Trerè Silvia Giovanelli Anna Miracolo Francesca Lodato marco montagnani Mariarosa Tamé Antonio Cloecchia Constance Mwangemi Davide Festi Enrico Roda Massimo Derenzini Giuseppe Mazzella Department of Internal Medicine and Gastroenterology University of Bologna Department of Experimental Pathology University of Bologna

AIM:To determine the role of interferon(IFN)with or withoutribavirin in preventing or delaying hepatocellular carcinoma(HCC)development in patients with hepatitis C virus(HCV)related *** on the preventive effect of IFN plusribavirin treatment are ***:A total of 101 patients(62 males and 39 females,mean age 55.1±1.4 years)with histologically proven HCVrelated liver cirrhosis plus compatible biochemistry andultrasonography were enrolled in the *** ultrasonography were performed every 6 *** liver biopsy was performed on all detected ***-up lasted for 5 *** proliferation,evaluated by measuring Ag-NOR proteins in hepatocytesnuclei,was expressed as AgNOR-Proliferative index(AgNOR-PI)(cut-off=2.5).Forty-one patients(27 males,14 females)were only followed up after the end of anyearly treatment with IFN-alpha2b(old treatment controlgroup=OTCG).Sixty naive patients were stratified accordingto sex and AgNOR-PI and then randomized in two groups:30 were treated with IFN-alpha2b+ribavirin(treatmentgroup=TG),the remaining were not treated(control group=CG).Nonresponders(NR)or relapsers in the TG receivedfurther IFN/ribavirin treatments after a 6 mo of ***:AgNOR-PI was significantly lowered by IFN(P2.5(26% vs3%,P<0.01).Two NR in the OTCG,none in the TG and 9 patients in the CG developed HCCduring *** Kaplan-Mayer survival curves showedstatistically significant differences both between OTCG andCG(P<0.004)and between TG and CG(P<0.003).CONCLUSION:IFN/ribavirin treatment associated with re-treatment courses of NR seems to produce the best resultsin terms of HCC ***-PI is a useful markerof possible HCC development.

关键词： Antineoplastic Agents dosage Antiviral Agents Carcinoma, Hepatocellular control Drug Therapy, Combination Female Hepatitis C, Chronic Humans Interferons Liver Cirrhosis Liver Neoplasms Male Middle Aged Nucleolus Organizer Region Prospective Studies Ribavirin Silver Staining

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Long-term alpha interferon and lamivudine combination therapy in non-responder patients with anti-HBe-positive chronic hepatitis B:Results of an open,controlled trial

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World Journal of Gastroenterology 2003年第7期9卷 1491-1495页

作者： M. Francesca Jaboli Carlo Fabbri Stefania Liva Francesco Azzaroli Giovanni Nigro Silvia Giovanelli Francesco Ferrara Anna Miracolo Sabrina Marchetto marco montagnani Antonio Colecchia Davide Festi Letizia Bacchi Reggiani Enrico Roda Giuseppe Mazzella Department of Internal Medicine and Gastroenterology University of BolognaItaly Department of Gastroenterology and Gastrointestinal Endoscopy Bellaria HospitalBolognaItaly Department of Medicine and Aging University of ChietiItaly Department of Cardiology University of BolognaItaly

AIM: To investigate the safety and efficacy of long-term combination therapy with alpha interferon and lamivudine in non-responsive patients with anti-HBe-positive chronic hepatitis ***: 34 patients received combination treatment (1 month lamivudine, 12 month lamivudine+interferon, 6month lamivudine), 24 received lamivudine (12 months),24 received interferon (12 months). Interferon was administered at 6 MU tiw and lamivudine at 100 mg orally once daily. Patients were followed up for 6 months after ***: At the end of treatment, HBV DNA negativity rates were 88 % with lamivudine+interferon, 99 % with lamivudine and 55 % with interferon, (P=0.004, combination therapy vs. interferon, and P=0.001 lamivudine ***), and serum transaminase normalization rates were 84 %, 91% and 53 % (P=0.01 combination therapy vs. interferon, and P=0.012 lamivudine vs. interferon). Six months later, HBV DNA negativity rates were 44 % with lamivudine+interferon, 33 % with lamivudine and 25 % with interferon, and serum transaminase normalization rates were 61%, 42 % and 45 %, respectively, without statistical significance. No YMDD variants were observed with lamivudine+interferon (vs. 12 % with lamivudine). The combination therapy appeared to be safe. CONCLUSION: Although viral clearance and transaminase normalization are slower with long-term lamivudine+interferon than that with lamivudine alone, the combination regimen seems to provide more lasting benefits and to protect against the appearance of YMDD variants. Studies with other regimens regarding sequence and duration are needed.

关键词： Adult Antiviral Agents dosage Drug Therapy, Combination Female Hepatitis B, Chronic Humans Interferon-alpha Lamivudine Male Middle Aged Reverse Transcriptase Inhibitors Treatment Outcome

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