-
Valeria Cento,
Simona Landonio,
Francesca De Luca,
Velia Chiara Di Maio, Valeria Micheli,
Carmen Mirabelli,
Fosca Niero,
Carlo Magni,
Giuliano Rizzardini,
Carlo Federico Perno,
Francesca Ceccherini-Silberstein
[show abstract]
[hide abstract]
ABSTRACT: A patient classified as HCV-1a positive by both LiPA Siemens 2.0 and Abbott RealTime HCV-Genotype II, was instead infected with HCV-1g, as determined by phylogenetic-analysis of NS3-sequences. HCV-1g NS3-sequences to date available naturally harbor the resistance-substitution T54S, plus P131S and L135F changes, located in the highly conserved NS3-positions within boceprevir-binding site, as determined by structural modeling. HCV-1g NS3-sequences show some similarities to HCV-4, poorly responsive to interferon/ribavirin and to boceprevir/telaprevir; this patient was also a null-responder to boceprevir-treatment. Baseline genotypic-resistance testing may provide critical information for the management of first-generation protease-inhibitors based regimens, including both HCV genotype/subtype and natural resistance.
Antiviral therapy 02/2013; · 3.16 Impact Factor
-
Valentina Svicher,
Valeria Cento,
Gabriella Rozera,
Isabella Abbate,
Maria Mercedes Santoro,
Daniele Armenia,
Lavinia Fabeni,
Alessandro Bruselles,
Alessandra Latini,
Guido Palamara, Valeria Micheli,
Giuliano Rizzardini,
Caterina Gori,
Federica Forbici,
Giuseppe Ippolito,
Massimo Andreoni,
Andrea Antinori,
Francesca Ceccherini-Silberstein,
Maria Rosaria Capobianchi,
Carlo Federico Perno
[show abstract]
[hide abstract]
ABSTRACT: The false-positive rate (FPR) is a percentage-score provided by Geno2Pheno-algorithm indicating the likelihood that a V3-sequence is falsely predicted as CXCR4-using. We evaluated the correlation between FPR obtained by V3 population-sequencing and the burden of CXCR4-using variants detected by V3 ultra-deep sequencing (UDPS) and Enhanced-Sensitivity Trofile assay (ESTA).
54 HIV-1 B-subtype infected-patients (all maraviroc-naïve), with viremia >10,000copies/ml, were analyzed. HIV-tropism was assessed by V3 population-sequencing, UDPS (considering variants with >0.5% prevalence), and ESTA.
By UDPS, CCR5-using variants were detected in 53/54 patients, irrespective of FPR values, and their intra-patient prevalence progressively increased by increasing the FPR obtained by V3 population-sequencing (rho = 0.75, p = 5.0e-8). Conversely, the intra-patient prevalence of CXCR4-using variants in the 54 patients analyzed progressively decreased by increasing the FPR (rho = -0.61; p = 9.3e-6). Indeed, no CXCR4-using variants were detected in 13/13 patients with FPR>60. They were present in 7/18 (38.8%) patients with FPR 20-60 (intra-patient prevalence range: 2.1%-18.4%), in 5/7 (71.4%) with FPR 10-20, in 4/6 (66.7%) with FPR 5-10, and in 10/10(100%) with FPR<5 (intra-patient prevalence range: 12.1%-98.1%).
FPR by V3 population-sequencing can predict the burden of CXCR4-using variants. This information can be used to optimize the management of tropism determination in clinical practice. Due to its low cost and short turnaround time, V3 population-sequencing may represent the most feasible test for HIV-1 tropism determination. More sensitive methodologies (as UDPS) might be useful when V3 population-sequencing provides a FPR >20 (particularly in the range 20-60), allowing a more careful identification of patients harboring CXCR4-using variants.
PLoS ONE 01/2013; 8(1):e53603. · 4.09 Impact Factor
-
Valeria Cento,
Carmen Mirabelli,
Salvatore Dimonte,
Romina Salpini,
Yue Han,
Pascale Trimoulet,
Ada Bertoli, Valeria Micheli,
Guido Gubertini,
Giuseppina Cappiello, [......],
Roberta Longo,
Martina Bernassola,
Francesco Mazzotta,
Giuseppe Maria De Sanctis,
Xin Xin Zhang,
Jens Verheyen,
Antonella D'Arminio Monforte,
Francesca Ceccherini-Silberstein,
Carlo Federico Perno,
Valentina Svicher
[show abstract]
[hide abstract]
ABSTRACT: How the overlapping between HBV RT- and HBsAg-gene modulates the extent of HBV genetic-variability is still an open question. The rate of nucleotide-conservation (<1% variability) followed an atypical pattern in RT-gene, due to RT/HBsAg-overlapping (69.9% vs. 41.1% non-overlapping; p<0.001), consequent to the lower rate of synonymous-substitution within overlapping-region (median[IQR]dS=3.1[1.5-7.4] vs. 20.1[10.6-30.0]; p=3.249•10-22). The most conserved RT-regions were located within the YMDD-motif and the N-terminal parts of the palm- and finger-domain, critical for RT-functionality. These regions also corresponded to highly conserved HBsAg-domains critical for HBsAg-secretion. Conversely, the genomic-region encoding the HBsAg antigenic-loop (where immune-escape mutations are localized) showed a sharp decrease in the extent of conservation (40.6%), less pronounced in the setting of HIV-driven immune-suppression (48.8% vs. 21.5% in mono-infected;p=0.020). In conclusion, the overlapping reading-frame and the immune-system have shaped the patterns of RT and HBsAg genetic-variability. Highly conserved regions in RT and HBsAg may deserve further attention as novel therapeutic-targets.
Journal of General Virology 10/2012; · 3.36 Impact Factor
-
Marco Franzetti,
Alessia Lai,
Francesco Roberto Simonetti,
Giorgio Bozzi,
Andrea De Luca, Valeria Micheli,
Paola Meraviglia,
Paola Corsi,
Patrizia Bagnarelli,
Paolo Almi,
Alessia Zoncada,
Claudia Balotta
[show abstract]
[hide abstract]
ABSTRACT: Transmitted drug resistance (TDR) is mainly restricted to individuals carrying B subtype, with low prevalence among non-B subtypes when grouped together. Subtype F1 is the most frequent non-B variant found in subjects living in Italy, allowing a specific assessment of TDR associated with this clade.
We analysed pol sequences of HIV-1-positive individuals carrying the F1 variant included in the Antiretroviral Resistance Cohort Analysis database in the 1998-2009 period. Mutations were analysed with the Surveillance Drug Resistance Mutation and the International AIDS Society lists for naive and treated patients, respectively.
Among 343 HIV-1-infected patients carrying an F1 subtype, resistance was evaluated in a subset of 221 patients whose treatment status was known (169 drug naive and 52 drug experienced). The prevalence of TDR was 15.4% (11.8% for nucleoside/nucleotide reverse transcriptase inhibitors, 6.5% for non-nucleoside reverse transcriptase inhibitors and 7.1% for protease inhibitors). Among the 169 naive patients, 75.1%, 10.1% and 7.1% were Italians, South Americans and Romanians, respectively. Heterosexuals were prevalent among Italians and Romanians, while men who have sex with men were predominant among South Americans. The overall frequency of TDR declined from 21.4% to 7.1% in the 1998-2009 period. Although no statistical difference was detected, the frequency of TDR was higher in South Americans (23.5%) compared with Italian and Romanian naive patients (15% and 8.3%, respectively).
Our study shows a remarkable frequency of TDR in the F1 subtype-infected population. The high prevalence of TDR detected in South American subjects is linked to the homosexual route of infection. However, TDR was considerably high also in Italian subjects harbouring the F1 subtype, deserving careful monitoring.
Journal of Antimicrobial Chemotherapy 02/2012; 67(5):1250-3. · 5.07 Impact Factor
-
Daniele Armenia,
Ina Vandenbroucke,
Lavinia Fabeni,
Herwig Van Marck,
Valeria Cento,
Roberta D'Arrigo,
Liesbeth Van Wesenbeeck,
Fernanda Scopelliti, Valeria Micheli,
Bianca Bruzzone,
Sergio Lo Caputo,
Jeroen Aerssens,
Giuliano Rizzardini,
Valerio Tozzi,
Pasquale Narciso,
Andrea Antinori,
Lieven Stuyver,
Carlo Federico Perno,
Francesca Ceccherini-Silberstein
[show abstract]
[hide abstract]
ABSTRACT: The dynamics of raltegravir-resistant variants and their impact on virologic response in 23 HIV-1-infected patients, who started a salvage raltegravir-containing regimen, were investigated.
Integrase population sequencing and Ultra-Deep-454 Pyrosequencing (UDPS) were performed on plasma samples at baseline and at raltegravir failure. All integrase mutations detected at a frequency ≥1% were considered to be reliable for the UDPS analyses. Phylogenetic and phenotypic resistance analyses were also performed.
At baseline, primary resistance mutations were not detected by both population and UDPS genotypic assays; few secondary mutations (T97A-V151I-G163R) were rarely detected and did not show any statistically association either with virologic response at 24-weeks or with the development of resistant variants at failure. At UDPS, not all resistant variants appearing early during treatment evolved as major populations during failure; only specific resistance pathways (Y143R-Q148H/R-N155H) associated with an increased rate of fitness and phenotypic resistance were selected.
Resistance to raltegravir in integrase strand transfer inhibitor-naive patients remains today a rare event, which might be changed by future extensive use of such drugs. In our study, pathways of resistance at failure were not predicted by baseline mutations, suggesting that evolution plus stochastic selection plays a major role in the appearance of integrase-resistance mutations, whereas fitness and resistance are dominant factors acting for the late selection of resistant quasispecies.
The Journal of Infectious Diseases 02/2012; 205(4):557-67. · 6.41 Impact Factor
-
Alessia Lai,
Francesco R Simonetti,
Gianguglielmo Zehender,
Andrea De Luca, Valeria Micheli,
Paola Meraviglia,
Paola Corsi,
Patrizia Bagnarelli,
Paolo Almi,
Alessia Zoncada,
Stefania Paolucci,
Angela Gonnelli,
Grazia Colao,
Danilo Tacconi,
Marco Franzetti,
Massimo Ciccozzi,
Maurizio Zazzi,
Claudia Balotta
[show abstract]
[hide abstract]
ABSTRACT: About 40% of the Italian HIV-1 epidemic due to non-B variants is sustained by F1 clade, which circulates at high prevalence in South America and Eastern Europe. Aim of this study was to define clade F1 origin, population dynamics and epidemiological networks through phylogenetic approaches. We analyzed pol sequences of 343 patients carrying F1 subtype stored in the ARCA database from 1998 to 2009. Citizenship of patients was as follows: 72.6% Italians, 9.3% South Americans and 7.3% Rumanians. Heterosexuals, Homo-bisexuals, Intravenous Drug Users accounted for 58.1%, 24.0% and 8.8% of patients, respectively. Phylogenetic analysis indicated that 70% of sequences clustered in 27 transmission networks. Two distinct groups were identified; the first clade, encompassing 56 sequences, included all Rumanian patients. The second group involved the remaining clusters and included 10 South American Homo-bisexuals in 9 distinct clusters. Heterosexual modality of infection was significantly associated with the probability to be detected in transmission networks. Heterosexuals were prevalent either among Italians (67.2%) or Rumanians (50%); by contrast, Homo-bisexuals accounted for 71.4% of South Americans. Among patients with resistant strains the proportion of clustering sequences was 57.1%, involving 14 clusters (51.8%). Resistance in clusters tended to be higher in South Americans (28.6%) compared to Italian (17.7%) and Rumanian patients (14.3%). A striking proportion of epidemiological networks could be identified in heterosexuals carrying F1 subtype residing in Italy. Italian Heterosexual males predominated within epidemiological clusters while foreign patients were mainly Heterosexual Rumanians, both males and females, and South American Homo-bisexuals. Tree topology suggested that F1 variant from South America gave rise to the Italian F1 epidemic through multiple introduction events. The contact tracing also revealed an unexpected burden of resistance in epidemiological clusters underlying the need of public interventions to limit the spread of non-B subtypes and transmitted drug resistance.
PLoS ONE 01/2012; 7(8):e42223. · 4.09 Impact Factor
-
Valentina Svicher,
Claudia Alteri,
Marco Montano,
Roberta D'Arrigo,
Massimo Andreoni,
Gioacchino Angarano,
Andrea Antinori,
Guido Antonelli,
Tiziano Allice,
Patrizia Bagnarelli, [......],
Renzo Scaggiante,
Gaetana Sterrantino,
Ombretta Turriziani,
Maria Linda Vatteroni,
Laura Vecchi,
Claudio Viscoli,
Vincenzo Vullo,
Maurizio Zazzi,
Adriano Lazzarini,
Carlo Federico Perno
[show abstract]
[hide abstract]
ABSTRACT: The DIVA study is aimed at setting up a standardized genotypic tropism-testing on proviral-DNA for the routine clinical diagnostic-laboratory.
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).
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 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.
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.
The new microbiologica: official journal of the Italian Society for Medical, Odontoiatric, and Clinical Microbiology (SIMMOC) 01/2012; 35(1):17-25. · 1.00 Impact Factor
-
Valentina Svicher,
Valeria Cento,
Martina Bernassola,
Maria Neumann-Fraune,
Formijn Van Hemert,
Mengjie Chen,
Romina Salpini,
Chang Liu,
Roberta Longo,
Michela Visca, [......],
Francesca Ceccherini-Silberstein,
Cesare Sarrecchia,
Massimo Andreoni,
Mario Angelico,
Antonella Ursitti,
Alberto Spanò,
Jing Maria Zhang,
Jens Verheyen,
Giuseppina Cappiello,
Carlo Federico Perno
[show abstract]
[hide abstract]
ABSTRACT: Occult HBV infection (OBI) is a threat for the safety of blood-supply, and has been associated with the onset of HBV-related hepatocellular carcinoma and lymphomagenesis. Nevertheless, genetic markers in HBsAg (particularly in D-genotype, the most common in Europe) significantly associated with OBI in vivo are missing. Thus, the goal of this study is to define: (i) prevalence and clinical profile of OBI among blood-donors; (ii) HBsAg-mutations associated with OBI; (iii) their impact on HBsAg-detection. OBI was searched among 422,278 blood-donors screened by Nucleic-Acid-Testing. Following Taormina-OBI-definition, 26 (0.006%) OBI-patients were identified. Despite viremia <50IU/ml, HBsAg-sequences were obtained for 25/26 patients (24/25 genotype-D). OBI-associated mutations were identified by comparing OBI-HBsAg with that of 82 chronically-infected (genotype-D) patients as control. Twenty HBsAg-mutations significantly correlated for the first time with OBI. By structural analysis, they localized in the major HBV B-cell-epitope, and in HBsAg-capsid interaction region. 14/24 OBI-patients (58.8%) carried in median 3 such mutations (IQR:2.0-6.0) against 0 in chronically-infected patients. By co-variation analysis, correlations were observed for R122P+S167L (phi=0.68, P=0.01), T116N+S143L (phi=0.53, P=0.03), and Y100S+S143L (phi=0.67, p<0.001). Mutants (obtained by site-directed mutagenesis) carrying T116N, T116N+S143L, R122P, R122P+Q101R, or R122P+S167L strongly decreased HBsAg-reactivity (54.9±22.6S/CO, 31.2±12.0S/CO, 6.1±2.4S/CO, 3.0±1.0S/CO and 3.9±1.3S/CO, respectively) compared to wild-type (306.8±64.1S/CO). Even more, Y100S and Y100S+S143L supernatants show no detectable-HBsAg (experiments in quadruplicate). In conclusions, unique HBsAg-mutations in genotype-D, different than those described in genotypes B/C (rarely found in western countries), tightly correlate with OBI, and strongly affect HBsAg-detection. By altering HBV-antigenicity and/or viral-particle maturation, they may affect full-reliability of universal diagnostic-assays for HBsAg-detection.
Antiviral research 11/2011; 93(1):86-93. · 3.61 Impact Factor
-
Alessia Lai,
Michela Violin,
Erika Ebranati,
Marco Franzetti, Valeria Micheli,
Maria Rita Gismondo,
Amedeo Capetti,
Paola Meraviglia,
Francesco Roberto Simonetti,
Giorgio Bozzi,
Masimo Ciccozzi,
Massimo Galli,
Gianguglielmo Zehender,
Claudia Balotta
[show abstract]
[hide abstract]
ABSTRACT: Transmission of HIV-1 and drug resistance continue to occur at a considerable level in Italy, influenced mainly by changes in modality of infection. However, the long period of infectivity makes difficult the interpretation of epidemiological networks, based on epidemiological data only. We studied 510 naive HIV-1-infected individuals, of whom 400 (78.4%) were newly diagnosed patients with an unknown duration of infection (NDs), with the aim of identifying sexual epidemiological networks and transmitted drug resistance (TDR) over a 7-year period. Clusters were identified by Bayesian methods for 412 patients with B subtype; 145 individuals (35.2%) clustered in 34 distinct clades. Within epidemiological networks males were 93.1% (n=135); the same proportion of patients has been infected by the sexual route; 62.1% (n=90) were men having sex with men (MSM) of whom 67.8% (n=61) were NDs. Among heterosexuals (n=44), males were predominant (79.5%, n=35) and 77.3% (n=34) were NDs. TDR in clusters was 11.7 % (n=17), of whom 76.5% (n=13) was found in MSM. TDR was predominantly associated with NRTI resistance in individuals with chronic infection (n=11). A high prevalence of epidemiological networks has been found in the metropolitan area of Milan, indicating a high frequency of transmission events. The cluster analysis of networks suggested that the source of new infections was mainly represented by males and MSM who have long lasting HIV-1 infection. Notably, the prevalence of resistance-conferring mutations was higher in chronically infected patients, carrying mainly resistance to thymidine analogs, the backbone of first antiretroviral (ARV) generation. Intervention strategies of public health are needed to limit HIV-1 transmission and the associated TDR.
AIDS research and human retroviruses 09/2011; 28(8):857-65. · 2.18 Impact Factor
-
Valentina Svicher,
Valeria Cento,
Romina Salpini,
Fabio Mercurio,
Maria Fraune,
Bastian Beggel,
Yue Han,
Caterina Gori,
Linda Wittkop,
Ada Bertoli, [......],
Francesco Mazzotta,
Giuseppe Maria De Sanctis,
Hervè Fleury,
Pascale Trimoulet,
Mario Angelico,
Giuseppina Cappiello,
Xin Xin Zhang,
Jens Verheyen,
Francesca Ceccherini-Silberstein,
Carlo Federico Perno
[show abstract]
[hide abstract]
ABSTRACT: Impact of hepatitis B virus genetic barrier, defined as the number and type of nucleotide substitutions required to overcome drug/immune selective pressure, on drug-resistance/immune-escape development is unknown.
Genetic barrier was calculated according to Van de Vijver (2006) in 3482 hepatitis B virus-reverse transcriptase/HBV surface antigen sequences from 555 drug-naïve patients and 2927 antiviral-treated patients infected with hepatitis B virus genotypes A-G.
Despite high natural variability, genetic barrier for drug-resistance development is identical amongst hepatitis B virus genotypes, but varies according to drug-resistance mutation type. Highest genetic barrier is found for secondary/compensatory mutations (e.g. rtL80I/V-rtL180M-rtV173L), whilst most primary mutations (including rtM204V-rtA181T/V-rtI169T-rtA194T) are associated with low genetic barrier. An exception is rtM204I, which can derive from a transition or a transversion. Genotypes A and G are more prone to develop immune/diagnostic-escape mutations sT114R and sG130N. Vaccine-escape associated sT131N-mutation is a natural polymorphism in both A and G genotypes.
Genetic barrier and reverse transcriptase/HBV surface antigen overlapping can synergistically influence hepatitis B virus drug-resistance/immune-escape development. The different immune-escape potential of specific hepatitis B virus genotypes could have important clinical consequences in terms of disease progression, vaccine strategies and correct HBV surface antigen detection.
Digestive and Liver Disease 08/2011; 43(12):975-83. · 3.05 Impact Factor
-
Mattia C F Prosperi,
Simona Di Giambenedetto,
Iuri Fanti,
Genny Meini,
Bianca Bruzzone,
Annapaola Callegaro,
Giovanni Penco,
Patrizia Bagnarelli, Valeria Micheli,
Elisabetta Paolini,
Antonio Di Biagio,
Valeria Ghisetti,
Massimo Di Pietro,
Maurizio Zazzi,
Andrea De Luca
[show abstract]
[hide abstract]
ABSTRACT: HIV-1 genotypic susceptibility scores (GSSs) were proven to be significant prognostic factors of fixed time-point virologic outcomes after combination antiretroviral therapy (cART) switch/initiation. However, their relative-hazard for the time to virologic failure has not been thoroughly investigated, and an expert system that is able to predict how long a new cART regimen will remain effective has never been designed.
We analyzed patients of the Italian ARCA cohort starting a new cART from 1999 onwards either after virologic failure or as treatment-naïve. The time to virologic failure was the endpoint, from the 90th day after treatment start, defined as the first HIV-1 RNA > 400 copies/ml, censoring at last available HIV-1 RNA before treatment discontinuation. We assessed the relative hazard/importance of GSSs according to distinct interpretation systems (Rega, ANRS and HIVdb) and other covariates by means of Cox regression and random survival forests (RSF). Prediction models were validated via the bootstrap and c-index measure.
The dataset included 2337 regimens from 2182 patients, of which 733 were previously treatment-naïve. We observed 1067 virologic failures over 2820 persons-years. Multivariable analysis revealed that low GSSs of cART were independently associated with the hazard of a virologic failure, along with several other covariates. Evaluation of predictive performance yielded a modest ability of the Cox regression to predict the virologic endpoint (c-index≈0.70), while RSF showed a better performance (c-index≈0.73, p < 0.0001 vs. Cox regression). Variable importance according to RSF was concordant with the Cox hazards.
GSSs of cART and several other covariates were investigated using linear and non-linear survival analysis. RSF models are a promising approach for the development of a reliable system that predicts time to virologic failure better than Cox regression. Such models might represent a significant improvement over the current methods for monitoring and optimization of cART.
BMC Medical Informatics and Decision Making 06/2011; 11:40. · 1.48 Impact Factor
-
Nicola Gianotti,
Laura Galli,
Maurizio Zazzi,
Valeria Ghisetti,
Stefano Bonora, Valeria Micheli,
Paola Meraviglia,
Paola Corsi,
Bianca Bruzzone,
Stefano Menzo,
Simona Di Giambenedetto,
Andrea De Luca,
Gaetano Filice,
Giovanni Penco,
Antonella Castagna
[show abstract]
[hide abstract]
ABSTRACT: An investigation was undertaken to determine whether specific pol mutations hinder long-term immune recovery regardless of virological response. In total, 826 patients with >50 HIV RNA copies/ml, who underwent genotypic resistance testing between 1 January 2000 and 31 December 2003 after >3 years of antiretroviral treatment, and were followed up for >3 years after genotypic resistance testing, were analyzed retrospectively. The outcome of the study was the lack of immune recovery after >3 years of follow-up, defined as a slope by linear regression <0. The viremia detectability ratio was defined as the number of HIV RNA values of >50 copies/ml divided by the number of HIV RNA measurements during follow-up. Logistic regression was used for univariable and multivariable analysis. Median (Q1, Q3) values at baseline were the following: age 40 (37, 45) years, years on antiretroviral therapy 4.45 (3.65, 5.47), HIV RNA 3.91 (3.39, 4.53) log(10) copies/ml, CD4+ T-cell 358 (211, 524)/µl. After 3.13 years of follow-up, 375 patients (45.4%) showed a lack of immune recovery. The risk of lack of immune recovery increased independently with increasing baseline CD4+ counts (OR=1.104 per 50-cell increase, 95% CI=1.069-1.142, P<0.0001), increasing viremia detectability ratio during follow-up (OR=1.145 per 0.1-unit increase, 95% CI=1.093-1.202, P<0.0001), and with earlier calendar years of resistance testing (overall effect: P=0.0007). In conclusion, no pol mutation is associated independently with the lack of immune recovery.
Journal of Medical Virology 03/2011; 83(3):391-8. · 2.82 Impact Factor
-
Andrea De Luca,
Simona Di Giambenedetto,
Renato Maserati,
Nicola Gianotti,
Pasquale Narciso,
Andrea Antinori,
Giovanni Di Perri,
Mattia C F Prosperi,
Fausto Baldanti, Valeria Micheli,
Maurizio Zazzi,
Carlo F Perno,
Maria M Santoro
[show abstract]
[hide abstract]
ABSTRACT: There is not yet consensus on interpretation of genotypic HIV-1 resistance to darunavir (DRV). We validated existing rules and a newly derived score.
Protease inhibitor (PI)-failing patients starting a DRV/ritonavir-based regimen, with available baseline resistance genotypes, were extracted from three Italian databases. Virological response (VR) was analysed between 4 and 32 follow-up weeks, defined as a drop from baseline HIV RNA of ≥2 log(10) or a value <50 copies/ml if the last measurement had been obtained at ≤12 weeks and as HIV RNA<50 copies/ml if it had been obtained at >12 weeks of follow-up. DRV/ritonavir resistance was interpreted by seven algorithms. A new weighted score (DRV-2009) was derived and validated, analysing associations of protease mutations with VR.
A total of 217 patients were analysed, with a mean (±sd) follow-up time of 17 (±9) weeks. At baseline, median HIV RNA was 4.26 log(10) copies/ml (IQR 3.11-5.03); VR was achieved in 135/217 (62%) patients. Adjusting for use of a new drug class, number of previous PIs experienced, CD4(+) T-cell count and HIV RNA, only the Rega DRV/ritonavir interpretation was significantly associated with VR (per increase in susceptibility category, OR 1.94, 95% CI 1.32-2.86; P<0.001). The DRV-2009 score V11I+L33F+R41K+I47V+2*I50V+2*I54M+K55R+D60E+L74P+L76V+N88D+2*L89V-L10I/V-I13V-G16E-G48V-F53I/L-I62V-I66F-V77I (<0 indicating susceptibility, 0-1 intermediate resistance and ≥2 resistance) correlated with VR in the derivation set (n=132, R=0.395; P<0.001). In the validation set (n=85), after adjusting for mutual interpretation and new use of enfuvirtide, DRV-2009 (P=0.017) and Rega (P=0.013) were both independently associated with VR.
In contrast to the other algorithms, both the DRV-2009 score and Rega interpretation showed a robust predictive capacity of VR to DRV/ritonavir-containing regimens.
Antiviral therapy 01/2011; 16(4):489-97. · 3.16 Impact Factor
-
Valentina Svicher,
Claudia Alteri,
Caterina Gori,
Romina Salpini,
Fabbio Marcuccilli,
Ada Bertoli,
Roberta Longo,
Martina Bernassola,
Valentina Gallinaro,
Sara Romano, [......], Valeria Micheli,
Mario Angelico,
Giovanni Cassola,
Giustino Parruti,
Guido Gubertini,
Giuseppe Maria De Sanctis,
Francesca Ceccherini-Silberstein,
Giuseppina Cappiello,
Alberto Spanò,
Carlo Federico Perno
[show abstract]
[hide abstract]
ABSTRACT: To investigate lamivudine (LAM)-resistance profiles of hepatitis B virus (HBV) at the early stages of virological breakthrough (serum HBV-DNA 12-345IU/ml) or when HBV-DNA is undetectable.
Sixty-four HBV-mono-infected patients were enrolled: 25 had virological breakthrough with serum HBV-DNA ranging from 12 to 345IU/ml during first-line LAM-monotherapy; 24 were on LAM-monotherapy, and 15 were on LAM+adefovir dipivoxil (ADV) with undetectable serum HBV-DNA (<12IU/ml).
HBV-reverse transcriptase was successfully sequenced in 22 (88.0%) LAM-treated patients with HBV-DNA between 12 and 345IU/ml, and in 12 (30.8%) patients receiving LAM (±ADV) with HBV-DNA<12IU/ml. Drug-resistance mutations were observed in 17 (77.2%) LAM-treated patients with virological breakthrough: 8 M204V, 7 M204I, 1 M204I/V, and 1 A181T. One or ≥2 compensatory mutations were found in 10 (58.8%) and in 4 (23.5%) patients. Drug-resistance mutations were present also in patients with undetectable serum HBV-DNA: M204I was detected in 2 patients receiving LAM-monotherapy, and V84M in 1 patient receiving LAM+ADV.
Overall findings support the existence of drug-resistance mutations even at very low levels of viral replication. The persistence of low-level HBV replication and consequent drug-resistance emergence should be considered when choosing therapeutic strategies.
Digestive and Liver Disease 12/2010; 42(12):902-7. · 3.05 Impact Factor
-
Valentina Svicher,
Roberta D'Arrigo,
Claudia Alteri,
Massimo Andreoni,
Gioacchino Angarano,
Andrea Antinori,
Guido Antonelli,
Patrizia Bagnarelli,
Fausto Baldanti,
Ada Bertoli, [......],
Giuliano Rizzardini,
Rosaria Santangelo,
Alessandro Soria,
Francesca Stazi,
Gaetana Sterrantino,
Ombretta Turriziani,
Claudio Viscoli,
Vincenzo Vullo,
Adriano Lazzarin,
Carlo Federico Perno
[show abstract]
[hide abstract]
ABSTRACT: 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 Trofile (ESTA) as reference-assay.
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%).
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.
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.
The new microbiologica: official journal of the Italian Society for Medical, Odontoiatric, and Clinical Microbiology (SIMMOC) 07/2010; 33(3):195-206. · 1.00 Impact Factor
-
Annalisa Saracino,
Laura Monno,
Alessandra Tartaglia,
Carmine Tinelli,
Elena Seminari,
Franco Maggiolo,
Stefano Bonora,
Stefano Rusconi, Valeria Micheli,
Sergio Lo Caputo,
Laura Lazzaroni,
Sergio Ferrara,
Nicoletta Ladisa,
Paola Nasta,
Giustino Parruti,
Rita Bellagamba,
Federica Forbici,
Gioacchino Angarano
[show abstract]
[hide abstract]
ABSTRACT: Tipranavir, a non-peptidic protease inhibitor which shows in vitro efficacy against some HIV-1-resistant strains, can be used in salvage therapies for multi-experienced HIV patients due to its peculiar resistance profile including 21 mutations at 16 protease positions according to International AIDS Society (IAS). Other genotypic scores, however, which attribute a different weight to single amino-acid substitutions, have been recently proposed. To validate the clinical utility of four different genotypic scores for selecting tipranavir responders, the baseline resistance pattern of 176 HIV heavily experienced patients was correlated with virological success (HIV-RNA<50 copies/ml) after 24 weeks of a new treatment based on tipranavir/ritonavir. Virological suppression after 24 weeks was reached by 42.5% of patients. With univariate analysis, genotypic scores were all associated with outcome but showed a low accuracy with ROC analysis, with the weighted score (WS) by Scherer et al. demonstrating the best performance with an AUC of 68%. Only 52% of patients classified as susceptible (WS< or =3) responded to the new therapy. The following variables were significantly associated (p<0.05) to failure with multivariate analysis: WS, log peak of HIV-RNA, IAS mutations: L33F, I54AMV, Q58E, and non-IAS mutation: N37DES. On the contrary, the use of T20 in T20-naïve patients and the V82AFSI and F53LY non-IAS mutations were associated with virological success. The study suggests that even if the "weighted" scores are able to interpret correctly the antiretroviral resistance profile of multi-experienced patients, it is difficult to individuate a cut-off which can be easily applied to this population for discriminating responders.
Current HIV research 07/2009; 7(4):425-33. · 1.98 Impact Factor
-
Annalisa Saracino,
Laura Monno,
Alessandra Tartaglia,
Carmine Tinelli,
Elena Seminari,
Franco Maggiolo,
Stefano Bonora,
Stefano Rusconi, Valeria Micheli,
Sergio L. Caputo,
Laura Lazzaroni,
Sergio Ferrara,
Nicoletta Ladisa,
Paola Nasta,
Giustino Parruti,
Rita Bellagamba,
Federica Forbici,
Gioacchino Angarano
[show abstract]
[hide abstract]
ABSTRACT: Tipranavir, a non-peptidic protease inhibitor which shows in vitro efficacy against some HIV-1-resistant strains, can be used in salvage therapies for multi-experienced HIV patients due to its peculiar resistance profile including 21 mutations at 16 protease positions according to International AIDS Society (IAS). Other genotypic scores, however, which attribute a different weight to single amino-acid substitutions, have been recently proposed. To validate the clinical utility of four different genotypic scores for selecting tipranavir responders, the baseline resistance pattern of 176 HIV heavily experienced patients was correlated with virological success (HIV-RNA<50 copies/ml) after 24 weeks of a new treatment based on tipranavir/ritonavir. Virological suppression after 24 weeks was reached by 42.5% of patients. With univariate analysis, genotypic scores were all associated with outcome but showed a low accuracy with ROC analysis, with the weighted score (WS) by Scherer et al. demonstrating the best performance with an AUC of 68%. Only 52% of patients classified as susceptible (WS®3) responded to the new therapy. The following variables were significantly associated (p<0.05) to failure with multivariate analysis: WS, log peak of HIV-RNA, IAS mutations: L33F, I54AMV, Q58E, and non-IAS mutation: N37DES. On the contrary, the use of T20 in T20-naïve patients and the V82AFSI and F53LY non-IAS mutations were associated with virological success. The study suggests that even if the “weighted” scores are able to interpret correctly the antiretroviral resistance profile of multi-experienced patients, it is difficult to individuate a cut-off which can be easily applied to this population for discriminating responders.
Current HIV Research 06/2009; 7(4):425-433. · 1.75 Impact Factor
-
Valentina Svicher,
Caterina Gori,
Maria Trignetti,
Michela Visca, Valeria Micheli,
Martina Bernassola,
Romina Salpini,
Guido Gubertini,
Roberta Longo,
Fosca Niero,
Francesca Ceccherini-Silberstein,
Giuseppe Maria De Sanctis,
Alberto Spanò,
Giuseppina Cappiello,
Carlo Federico Perno
[show abstract]
[hide abstract]
ABSTRACT: To investigate the different clusters of mutations associated with lamivudine resistance in HBV genotypes D and A.
HBV reverse transcriptase sequences of 89 HBV-infected patients failing lamivudine treatment were analyzed. The association of mutations with HBV genotypes was assessed by Chi-Squared test and multivariate logistic regression analysis. Covariate analysis was based on hierarchical clustering.
In genotype A, the rtM204V (prevalence: 68.2%) was the main sign of lamivudine failure. Multivariate analysis confirmed that genotype A is the only predictor for rtM204V emergence (OR: 14.5 [95% CI: 1.3-158], P=0.02). Covariate analysis showed that rtM204V clusters with rtL180M, rtL229V (corresponding to sF220L in the HBsAg), and, interestingly, with HBsAg mutation sS207N (bootstrap=0.95). Both sF220L and sS207N co-localized in the fourth transmembrane HBsAg domain. In contrast, in genotype D the primary mutations rtM204V and rtM204I occurred with similar prevalence (39.1% versus 45.3%, P=0.47), and showed a distinct pattern of compensatory mutations. rtM204V clusters with mutations localized in the RT-B domain (rtV173L, rtL180M, and rtT184A/S) (bootstrap=0.94), while rtM204I clusters with mutations localized in the RT-A domain (rtS53N, rtT54Y, and rtL80I/V) (bootstrap=0.96) (without associations with HBsAg specific mutations).
HBV genotype plays an important role in driving RT evolution under lamivudine treatment, and thus can be relevant for therapeutic sequencing, immunological response and disease progression.
Journal of Hepatology 11/2008; 50(3):461-70. · 9.26 Impact Factor
-
Valeria Micheli,
Mario Regazzi,
Laura Dickinson,
Paola Meraviglia,
Paola Villani,
Saye H Khoo,
Paolo Viganò,
Laura Cordier,
Maria Cusato,
Piergiorgio Duca,
Giovanna Orlando,
Giuliano Rizzardini,
David J Back,
Antonietta Cargnel
[show abstract]
[hide abstract]
ABSTRACT: Liver disease may alter the pharmacokinetics of antiretrovirals and produce changes in plasma protein binding. The aim was to evaluate the pharmacokinetics of total and unbound lopinavir (LPV) in HIV-infected patients with and without hepatitis C virus (HCV) coinfection. Fifty-six HIV+ patients receiving lopinavir/ritonavir (LPV/r) (group I = 24 controls; II = 23 HIV/HCV-coinfected; III = 9 cirrhotic HIV/HCV-coinfected) were included. Total (n = 56) and unbound (n = 36) LPV pharmacokinetic parameters were determined at steady-state using validated high-performance liquid chromatography with ultraviolet detection and high-performance liquid chromatography-tandem mass spectrometry methods, respectively. Pharmacokinetic parameters (plasma concentration just before drug administration, peak concentrations in plasma, times to maximum plasma concentration, areas under the plasma concentration-time curve from 0 to 12 hours, and CL/F/kg) of both total and unbound LPV were calculated by standard noncompartmental methods and differences among groups evaluated (Kruskal-Wallis test).LPV apparent oral clearance normalized to body weight (median, interquartile range) was 55 (40-68), 59 (44-69), and 71 (53-78) mL/h/kg for groups I, II, and III, respectively (II vs. I, P = 0.52; III vs. I, P = 0.16). The areas under the plasma concentration-time curve from 0 to 12 hours were 110.4 (80.9-135.2), 103.4 (85.5-131.3), and 92.8 (87.4-116.3) microg h/mL for groups I, II, and III, respectively (II vs. I, P = 0.68; III vs. I, P = 0.71). Chronic liver impairment produced a slight, although not significant, decrease in plasma protein binding. The free-fraction of LPV increased ( approximately 21%) from 0.97% (0.80-1.06) in HIV+/HCV- patients to 1.18% (0.89-1.65) in HIV/HCV+ cirrhotic patients. The apparent oral clearance of unbound LPV (CLu/F/kg) in cirrhotic patients did not change significantly, supporting the concept that the clearance of unbound LPV in liver disease is not affected after being inhibited by low-dose ritonavir co-administration.LPV total and unbound pharmacokinetics were not affected by hepatic impairment, suggesting that no adjustment of LPV/r dose is required for HIV/HCV-coinfected patients with and without cirrhosis and moderate impairment of liver function.
Therapeutic Drug Monitoring 06/2008; 30(3):306-13. · 2.49 Impact Factor
-
Stefano Rusconi,
Nicola Gianotti,
Fulvio Adorni,
Enzo Boeri,
Stefano Menzo,
Angela Gonnelli, Valeria Micheli,
Paola Meraviglia,
Michele Trezzi,
Elisabetta Paolini,
Andrea Giacometti,
Paola Corsi,
Massimo Di Pietro,
Laura Monno,
Grazia Punzi,
Maurizio Zazzi
JAIDS Journal of Acquired Immune Deficiency Syndromes 12/2007; 46(3):373-5. · 4.43 Impact Factor
