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51. Performance of genotypic tropism testing on proviral DNA in clinical practice: results from the DIVA study group

Author

Svicher, V, Alteri, C, Montano, M, D'Arrigo, R, Andreoni, M, Angarano, G, Antinori, A, Antonelli, G, Allice, T, Bagnarelli, P, Baldanti, F, Bertoli, A, Borderi, M, Boeri, E, Bon, I, Bruzzone, B, Callegaro, A, Capobianchi, M, Carosi, G, Cauda, R, Ceccherini Silberstein, F, Clementi, M, Chirianni, A, Colafigli, M, D'Arminio Monforte, A, De Luca, A, Di Biagio, A, Di Nicuolo, G, Di Perri, G, Di Pietro, M, Di Santo, F, Fabeni, L, Fadda, G, Galli, M, Gennari, W, Ghisetti, V, Giacometti, A, Gori, C, Gori, A, Gulminetti, R, Leoncini, F, Maffongelli, G, Maggiolo, F, Manca, G, Gargiulo, F, Martinelli, C, Maserati, R, Mazzotta, F, Meini, G, Micheli, V, Monno, L, Mussini, C, Narciso, P, Nozza, S, Paolucci, S, Pal, G, Parisi, S, Parruti, G, Pignataro, A, Pollicita, M, Quirino, T, Re, M, Rizzardini, G, Santangelo, R, Scaggiante, R, Sterrantino, G, Turriziani, O, Vatteroni, M, Vecchi, L, Viscoli, C, Vullo, V, Zazzi, M, Lazzarini, A, Perno, C, GORI, ANDREA, Perno, C., Svicher, V, Alteri, C, Montano, M, D'Arrigo, R, Andreoni, M, Angarano, G, Antinori, A, Antonelli, G, Allice, T, Bagnarelli, P, Baldanti, F, Bertoli, A, Borderi, M, Boeri, E, Bon, I, Bruzzone, B, Callegaro, A, Capobianchi, M, Carosi, G, Cauda, R, Ceccherini Silberstein, F, Clementi, M, Chirianni, A, Colafigli, M, D'Arminio Monforte, A, De Luca, A, Di Biagio, A, Di Nicuolo, G, Di Perri, G, Di Pietro, M, Di Santo, F, Fabeni, L, Fadda, G, Galli, M, Gennari, W, Ghisetti, V, Giacometti, A, Gori, C, Gori, A, Gulminetti, R, Leoncini, F, Maffongelli, G, Maggiolo, F, Manca, G, Gargiulo, F, Martinelli, C, Maserati, R, Mazzotta, F, Meini, G, Micheli, V, Monno, L, Mussini, C, Narciso, P, Nozza, S, Paolucci, S, Pal, G, Parisi, S, Parruti, G, Pignataro, A, Pollicita, M, Quirino, T, Re, M, Rizzardini, G, Santangelo, R, Scaggiante, R, Sterrantino, G, Turriziani, O, Vatteroni, M, Vecchi, L, Viscoli, C, Vullo, V, Zazzi, M, Lazzarini, A, Perno, C, GORI, ANDREA, and Perno, C.

Abstract

The DIVA study is aimed at setting up a standardized genotypic tropism-testing on proviral-DNA for the routine clinical diagnostic-laboratory. Methods: Twelve local centres and 5 reference centres (previously cross-validated) were identified. For inter-center validation-procedure, 60 peripheral-blood mononuclear cells (PBMCs) aliquots from 45 HAART-treated patients were randomly chosen for population V3 sequencing on proviral-DNA at local HIV centre and at reference-laboratory. Viral tropism was predicted by Geno2Pheno algorithm (False Positive Rate [FPR] = 20%) as proposed by the European-Guidelines. Quantification of total HIV-1 DNA was based on a method described by Viard (2004). Results: Quantification of HIV-1 DNA was available for 35/45 (77.8%) samples, and gave a median value of 598 (IQR:252- 1,203) copies/10 6 PBMCs. A total of 56/60 (93.3%) samples were successfully amplified by both the reference and the local virological centers. The overall concordance of tropism prediction between local and reference centers was 54/56 (96.4%). Results of tropism prediction by local centers were: 33/54 (61.1%) R5 and 21/54 (38.9%) X4/DM. Conclusion: There was high concordance in the genotypic tropism prediction based on proviral DNA among different virological centers throughout Italy. Our results are in line with other European studies, and support the use of genotypic tropism testing on proviral DNA in patients with suppressed plasma HIV-1 RNA candidate to CCR5-antagonist treatment.

Published
2012

57. Performance of genotypic tropism testing in clinical practice using the enhanced sensitivity version of Trofile as reference assay: results from the OSCAR Study Group

Author

Svicher, V, D'Arrigo, R, Alteri, C, Andreoni, M, Angarano, G, Antinori, A, Antonelli, G, Bagnarelli, P, Baldanti, F, Bertoli, A, Borderi, M, Boeri, E, Bonn, I, Bruzzone, B, Callegaro, Ap, Cammarota, R, Canducci, F, Ceccherini Silberstein, F, Clementi, M, Monforte, Ad, De Luca, Andrea, Di Biagio, A, Di Giambenedetto, Simona, Di Perri, G, Di Pietro, M, Fabeni, L, Fadda, Giovanni, Galli, M, Gennari, W, Ghisetti, V, Giacometti, A, Gori, A, Leoncini, F, Maggiolo, F, Maserati, R, Mazzotta, F, Micheli, V, Meini, G, Monno, L, Mussini, C, Nozza, S, Paolucci, S, Parisi, S, Pecorari, M, Pizzi, D, Quirino, T, Re, Mc, Rizzardini, G, Santangelo, Rosaria, Soria, A, Stazi, F, Sterrantino, G, Turriziani, O, Viscoli, C, Vullo, V, Lazzarin, A, Perno, Cf, Oscar, Study Group, De Luca, Andrea (ORCID:0000-0002-8311-6935), Di Giambenedetto, Simona (ORCID:0000-0001-6990-5076), Santangelo, Rosaria (ORCID:0000-0002-8056-218X), Svicher, V, D'Arrigo, R, Alteri, C, Andreoni, M, Angarano, G, Antinori, A, Antonelli, G, Bagnarelli, P, Baldanti, F, Bertoli, A, Borderi, M, Boeri, E, Bonn, I, Bruzzone, B, Callegaro, Ap, Cammarota, R, Canducci, F, Ceccherini Silberstein, F, Clementi, M, Monforte, Ad, De Luca, Andrea, Di Biagio, A, Di Giambenedetto, Simona, Di Perri, G, Di Pietro, M, Fabeni, L, Fadda, Giovanni, Galli, M, Gennari, W, Ghisetti, V, Giacometti, A, Gori, A, Leoncini, F, Maggiolo, F, Maserati, R, Mazzotta, F, Micheli, V, Meini, G, Monno, L, Mussini, C, Nozza, S, Paolucci, S, Parisi, S, Pecorari, M, Pizzi, D, Quirino, T, Re, Mc, Rizzardini, G, Santangelo, Rosaria, Soria, A, Stazi, F, Sterrantino, G, Turriziani, O, Viscoli, C, Vullo, V, Lazzarin, A, Perno, Cf, Oscar, Study Group, De Luca, Andrea (ORCID:0000-0002-8311-6935), Di Giambenedetto, Simona (ORCID:0000-0001-6990-5076), and Santangelo, Rosaria (ORCID:0000-0002-8056-218X)

Published
2010

59. Genotypic testing on HIV-1 DNA as a tool to assess HIV-1 co-receptor usage in clinical practice: results from the DIVA study group

Author

Svicher, V., primary, Alteri, C., additional, Montano, M., additional, Nori, A., additional, D’Arrigo, R., additional, Andreoni, M., additional, Angarano, G., additional, Antinori, A., additional, Antonelli, G., additional, Allice, T., additional, Bagnarelli, P., additional, Baldanti, F., additional, Bertoli, A., additional, Borderi, M., additional, Boeri, E., additional, Bon, I., additional, Bruzzone, B., additional, Barresi, R., additional, Calderisi, S., additional, Callegaro, A. P., additional, Capobianchi, M. R., additional, Gargiulo, F., additional, Castelli, F., additional, Cauda, R., additional, Ceccherini-Silberstein, F., additional, Clementi, M., additional, Chirianni, A., additional, Colafigli, M., additional, D’Arminio Monforte, A., additional, De Luca, A., additional, Di Biagio, A., additional, Di Nicuolo, G., additional, Di Perri, G., additional, Di Santo, F., additional, Fadda, G., additional, Galli, M., additional, Gennari, W., additional, Ghisetti, V., additional, Costantini, A., additional, Gori, A., additional, Gulminetti, R., additional, Leoncini, F., additional, Maffongelli, G., additional, Maggiolo, F., additional, Maserati, R., additional, Mazzotta, F., additional, Meini, G., additional, Micheli, V., additional, Monno, L., additional, Mussini, C., additional, Nozza, S., additional, Paolucci, S., additional, Palù, G., additional, Parisi, S., additional, Parruti, G., additional, Pignataro, A. R., additional, Quirino, T., additional, Re, M. C., additional, Rizzardini, G., additional, Sanguinetti, M., additional, Santangelo, R., additional, Scaggiante, R., additional, Sterrantino, G., additional, Turriziani, O., additional, Vatteroni, M. L., additional, Viscoli, C., additional, Vullo, V., additional, Zazzi, M., additional, Lazzarin, A., additional, and Perno, C. F., additional

Published
2013
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65. Changes in host cell molecules acquired by circulating HIV-1 in patients treated with highly active antiretroviral therapy and interleukin-2.

Author

Abbate, I, Dianzani, F, Turriziani, O, Antonelli, G, D'Offizi, G, Galati, V, Pierdominici, M, Pandolfi, Franco, Capobianchi, Mr, Pandolfi, Franco (ORCID:0000-0001-8799-8173), Abbate, I, Dianzani, F, Turriziani, O, Antonelli, G, D'Offizi, G, Galati, V, Pierdominici, M, Pandolfi, Franco, Capobianchi, Mr, and Pandolfi, Franco (ORCID:0000-0001-8799-8173)

Published
2001

69. Impaired 2′,3′-dideoxy-3′-thiacytidine accumulation in T-lymphoblastoid cells as a mechanism of acquired resistance independent of multidrug resistant protein 4 with a possible role for ATP-binding cassette C11

Author

TURRIZIANI, O., primary, SCHUETZ, J.D., additional, FOCHER, F., additional, SCAGNOLARI, C., additional, SAMPATH, J., additional, ADACHI, M., additional, BAMBACIONI, F., additional, RIVA, E., additional, and ANTONELLI, G., additional

Published
2002
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82. Cellular issues relating to the resistance of HIV to antiretroviral agents.

Author

Turriziani O, Scagnolari C, Bellomi F, Solimeo I, Focher F, and Antonelli G

Abstract

It has been proposed that the declining efficiency of antiretroviral agents in human immunodeficiency virus (HIV) infection may also depend on cellular factors at their site of action. Two in particular have been proposed: (i) the defective intracellular metabolism of NRTI in target cells and the altered uptake; and (ii) efflux of nucleoside reverse transcriptase inhibitors (NRTI) and protease inhibitors (PI) by cellular transporter molecules. Several studies have shown that: changes in the activities of various purine and pyrimidine biosynthetic enzymes may occur in lymphocytes of HIV-infected patients; HIV-infected patients on prolonged treatment with nucleoside analogues, e.g. zidovudine, show significantly decreased activity of thymidine kinase (TK) compared with untreated HIV-infected people; and NRTI and PI are substrates for the multidrug membrane transporters. With regard to the latter issue, it is known that the ATP-binding cassette transporter proteins such as the P-glycoprotein (MDR), and the newly discovered family of multidrug resistance-associated proteins (MRP1-6), promote the active extracellular efflux of a wide variety of therapeutics drugs and overexpression of some of them lowers intracellular concentration of PI. In the very near future such mechanisms, also called 'cellular drug resistance', might be taken into account, together with other immunological, virological and behavioural factors, to explain the 'drug failure' and/or the variability of response in HIV patients undergoing antiretroviral treatment. [ABSTRACT FROM AUTHOR]

Published
2003
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88. In vitroSelection of Human Immunodeficiency Virus Type 1 Resistant to Ro 31-8959 Proteinase Inhibitor

Author

Dianzani, F., Antonelli, G., Turriziani, O., Riva, E., Dong, G., and Bellarosa, D.

Abstract

Human immunodeficiency virus type I (HIV-1) resistant to the proteinase inhibitor Ro 31–8959 has been obtained by growing HIV-IIIB in C8166 cell cultures in the presence of inhibitory concentrations of the drug.Ro 31-8959-resistant HIV-1 obtained after five passages in the presence of the drug was capable of replicating, as measured by infectious virus yield, in the presence of concentrations of Ro 31-8959 able to inhibit completely the replication of the parental HIV-IIIB.Cloning of the Ro 31–8959-resistant viral preparations revealed that a number of different strains of HIV-1 with various degrees of sensitivity to Ro 31–8959 emerged during propagation of HIV-IIIB in C8166 cells in the presence of the drug. The most resistant strain shows an ID50value of 21.8 nM that is 13.2 fold higher than the value observed in HIV-IIIB infected C8166 cells.

Published
1993
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90. Performance of genotypic tropism testing in clinical practice using the enhanced sensitivity version of Trofile as reference assay: Results from the OSCAR Study Group

Author

Svicher V, D'Arrigo R, Alteri C, Andreoni M, Angarano G, Antinori A, Antonelli G, Bagnarelli P, Baldanti F, Bertoli A, Boeri E, Bruzzone B, Callegaro AP, Cammarota R, Canducci F, Ceccherini Silberstein F, Clementi M, Monforte AD, De Luca A, Di Biagio A, Di Gianbenedetto S, Di Perri G, Di Pietro M, Fabeni L, Fadda G, Galli M, Gennari W, Ghisetti V, Giacometti A, Gori A, Leoncini F, Maggiolo F, Maserati R, Mazzotta F, Micheli V, Meini G, Monno L, Mussini C, Nozza S, Paolucci S, Parisi S, Pecorari M, Pizzi D, Quirino T, Rizzardini G, Santangelo R, Soria A, Stazi F, Sterrantino G, Turriziani O, Viscoli C, Vullo V, Lazzarin A, Perno CF, OSCAR Study Group, BORDERI, MARCO, BON, ISABELLA, RE, MARIA CARLA, Svicher V, D'Arrigo R, Alteri C, Andreoni M, Angarano G, Antinori A, Antonelli G, Bagnarelli P, Baldanti F, Bertoli A, Borderi M, Boeri E, Bonn I, Bruzzone B, Callegaro AP, Cammarota R, Canducci F, Ceccherini-Silberstein F, Clementi M, Monforte AD, De Luca A, Di Biagio A, Di Gianbenedetto S, Di Perri G, Di Pietro M, Fabeni L, Fadda G, Galli M, Gennari W, Ghisetti V, Giacometti A, Gori A, Leoncini F, Maggiolo F, Maserati R, Mazzotta F, Micheli V, Meini G, Monno L, Mussini C, Nozza S, Paolucci S, Parisi S, Pecorari M, Pizzi D, Quirino T, Re MC, Rizzardini G, Santangelo R, Soria A, Stazi F, Sterrantino G, Turriziani O, Viscoli C, Vullo V, Lazzarin A, Perno CF, and OSCAR Study Group.

Subjects
Male, Protein Structure, trafile, Receptors, CCR5, Genotype, Settore MED/17 - Malattie Infettive, HIV, viral load, AIDS, hiv, HIV Infections, HIV Envelope Protein gp120, v3 loop, Receptors, Humans, Viral Tropism, Protein Structure, Tertiary, HIV-1, Female, Receptors, Virus, tropism, virus diseases, genotypic tropism testing, OSCAR Study Group, Settore MED/07 - Microbiologia e Microbiologia Clinica, Virus, genotypic tropism, CCR5 Receptor Antagonists, CCR5, Tertiary
Abstract

Objective: The goal of the OSCAR programme is to evaluate the performances of genotypic HIV-1 tropism testing in clinical practice using the enhanced sensitivity version of Trafile (ESTA) as reference-assay. Methods: HIV-1 coreceptor-usage was assessed using plasma samples from 406 HIV-1 infected patients by ESTA and by gp120 V3 population-sequencing followed by Geno2pheno (set at a False Positive Rate [FPR] of 10% and 5%). Results: ESTA was successful in 365 (89.9%) samples indicating R5 in 254 (69.6%), and DM/X4 in 111 (30.4% of samples (104 [28.5%] DM and 7 [1.9%] X4). Genotypic-testing successfully assessed viral tropism for all 406 samples, including the 41 with undetermined result by ESTA. Genotypic-tropism testing at a FPR of 5% and 10% was 81.1% and 78.4% concordant with ESTA, respectively. Despite a sensitivity of 48.7% and 55.9% at a FPR of 5% and 10%, respectively, a high concordance (specificity: 95.3% for FPR of 5% and 88.2% for FPR of 10%) between genotypic-tropism testing and ESTA was reached in the detection of R5-tropic viruses. Conclusion: Our results are in line with other European studies, and support the routine use of genotypic tropism testing in clinical-settings for monitoring of HIV-1 infected patients candidate to or failing CCR5-antagonists.

91. Alteration of thymidine kinase activity in cells treated with an antiviral agent

Author

Turriziani, O., Antonelli, G., Verri, G., Tozzi, V., Ferri, F., ELISABETTA RIVA, Bambacioni, E., and Dianzani, E.

Subjects
Anti-HIV Agents, Drug Resistance, hiv, HIV Infections, cellular resistance, In Vitro Techniques, Thymidine Kinase, aids, azt, thymidine kinase, Dideoxynucleosides, Cell Line, Kinetics, Leukocytes, Mononuclear, Humans, Phosphorylation, Zidovudine, Thymidine
Abstract

A lymphoblastoid cell line, CEM, was rendered resistant to zidovudine (AZT) in vitro by exposure to low but gradually increasing concentrations of the drug. This type of cellular resistance seems to be due to a defect of thymidine kinase (TK) activity that is acquired by cells grown in the presence of AZT. In fact, enzymatic studies with extracts from AZT-resistant cells (CEMazt), have shown that the value of the maximum velocity (Vmax) of TK activity measured with AZT and for deoxythymidine (dThd) is decreased as compared to sensitive CEM cells. Furthermore, the enzyme affinity for AZT and dThd is reduced in CEMazt. Further experiments have shown that such cells do not show resistance to other nucleoside analogs, such as ddI, ddC, AraT and D4T, suggesting that the phosphorylation pathways different from those involving TK are unaltered. Ex vivo experiments performed by using peripheral blood mononuclear cells (PBMC) from HIV infected individuals revealed that a prolonged treatment with AZT may modify the affinity of TK for dThd, thus suggesting that the aforementioned phenomenon may occur also in vivo.

92. Treatment with natural IFN of hepatitis C patients with or without antibodies to recombinant IFN

Author

Laura MONNO, Antonelli, G., Santantonio, T., Giannelli, Gianluigi, Currenti, M., Monno, L., Turriziani, O., Pastore, G., and Dianzani, F.

Subjects
Male, Time Factors, Interferon-alpha, Interferon alpha-2, Middle Aged, Antiviral Agents, Hepatitis C, Antibodies, Recombinant Proteins, Neutralization Tests, Case-Control Studies, Antibody Formation, Humans, Female, Hepatitis, Chronic
Abstract

The purpose of this study was to evaluate if a retreatment with natural human interferon alpha could lead to a recovery of the therapeutic responsiveness in non-responder patients with chronic hepatitis C who did (9 cases) or did not (14 cases) develop anti-interferon neutralizing antibodies while on treatment with recombinant interferon alpha2a. During retreatment, no patient developed detectable levels of neutralizing antibodies to natural interferon. At the end of retreatment, 6/9 positive patients showed a complete response to natural interferon therapy, while only 1/14 negative patients had a partial response. These data suggest that a second course of treatment with the natural preparation may be useful in patients who failed to respond to an earlier course of recombinant interferon, particularly the anti-interferon positive patients who showed hepatitis reactivation after an initial response concomitantly with antibody appearance. Conversely, a second course of natural interferon therapy might be useless in cases in which the lack of response is not associated to antibody development.

93. Evaluation of performances of VERSANT HCV RNA 1.0 assay (kPCR) and roche COBAS AmpliPrep/COBAS TaqMan HCV Test v2.0 at low level viremia

Author

Mazzuti, L., Lozzi, M. A., Riva, E., Maida, P., Falasca, F., Guido ANTONELLI, and Turriziani, O.

Subjects
VERSANT HCV RNA, HCV, Humans, RNA, Viral, COBAS TaqMan HCV, Real-time PCR, Viral load, Hepacivirus, Viremia, Hepatitis C, Polymerase Chain Reaction
Abstract

We assess the concordance between low level HCV values obtained using the VERSANT HCV RNA 1.0 Assay (kPCR) and COBAS AmpliPrep/COBAS TaqMan HCV Quantitative Test v2.0. The correlation between the values obtained by the two RT-PCR assays for samples with quantifiable HCV RNA levels revealed that viral load measured by kPCR significantly correlated with that of the CAP/CTM (R=0.644, P0.0001). The results show a good concordance (n=126/144, 87%); discordant results were mainly observed in the assessment of values below the lower limit of detection of the assays. These variations may have an impact on clinical decisions for patients on HCV triple therapy or interferon- free regimens. It is therefore recommended to monitor individual patients with the same test throughout treatment.

94. Identification of an amino acid substitution involved in the reduction of sensitivity of HIV-1 to an inhibitor of viral proteinase

Author

Turriziani, O., Antonelli, G., Jacobsen, H., Mous, J., ELISABETTA RIVA, Pistello, M., and Dianzani, F.

Subjects
Base Sequence, Molecular Sequence Data, Glycine, HIV, Drug Resistance, Microbial, Valine, HIV Protease Inhibitors, Isoquinolines, Viral Proteins, Mutation, HIV-1, Quinolines, Humans, Saquinavir, DNA Primers
Abstract

Clones derived from HIV variants previously characterized as resistant to Ro31-8959, an inhibitor of viral proteinase (PR), were sequenced. Substitution of glycine by valine at position 48 of the PR protein was found. None of the 20 clones derived from wild type HTLV-IIIB contain this mutation. Since such a position is located in a conserved region of PR, it is possible that the substitution can affect the interaction of the enzyme with the inhibitor.

95. Genetic analyses of HIV-1 pol sequences from Zimbabwean patients

Author

Falasca, F., Temperoni, C., Mazzuti, L., Presti, A. L., massimo ciccozzi, Di Filippo, E., Iaiani, G., Taliani, G., and Turriziani, O.

Subjects
Adult, Male, zimbabwe, pol sequence, phylogeny, drug resistance, Adolescent, Molecular Sequence Data, Genetic Variation, HIV Infections, Middle Aged, Young Adult, pol Gene Products, Human Immunodeficiency Virus, Child, Preschool, Mutation, HIV-1, Humans, Female
Abstract

This study analysed the sequence of HIV-1 pol gene derived from Zimbabwean infected patients. Sequence analysis, performed on 8 samples, revealed that sequences were classified as subtype C (n=5), subtype B (n=2) and CRF01_AE (n=1). Two patients, treated with a therapeutic regimen containing NRTI/NNRTI, harboured drug resistance mutations in HIV-1 DNA. Phylogenetic analysis performed on subtype C sequences showed that our strains were aggregated in different clusters depending on the country of origin.

100. Drug combination of AZT and ddI - synergism of action and prevention of appearance of AZT-resistance

Author

Antonelli, G., Dianzani, F., Bellarosa, D., Turriziani, O., Riva, E., and Gentile, A.

Subjects
Zidovudine -- Physiological aspects, Didanosine -- Physiological aspects, Drug resistance in microorganisms -- Prevention, Drug synergism -- Research
Abstract

SOURCE: Antiviral Chemistry & Chemotherapy, January 1994;5(1):51-55. According to the authors' abstract of an article published in Antiviral Chemistry & Chemotherapy, "Both 3'-azido-3'-deoxythymidine (AZT) and 2',3'-dideoxyinosine (ddI) strongly inhibit the [...]

Published
1994

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