bluuebunny/crossref_metadata_2025 huggingface dataset:
Selection of Hepatitis B Virus Polymerase Mutations in HIV-Coinfected Patients Treated with Tenofovir
Authors: Sheldon Julie , Camino Nuria , Rodés Berta , Bartholomeusz Angeline , Kuiper Michael , Tacke Frank , Núñez Marina , Mauss Stefan , Lutz Thomas , Klausen Gerd , Locarnini Stephen , Soriano Vincent
Published: April 2022

Background

Tenofovir (TDF) is an adenosine nucleotide analogue that has been approved for the treatment of HIV-1 infection. It also shows activity against hepatitis B virus (HBV) in patients with or without lamivudine (LAM)-associated mutations. Development of clinical or virological HBV breakthrough during TDF therapy has not been reported so far. The aim of this study was to analyse the HBV polymerase (pol) from HIV/HBV-coinfected patients with detectable serum levels of HBV DNA during treatment with TDF for longer than 6 months.

Methods

The HBV pol was sequenced from 43 patient's serum before and during TDF therapy. Phenotypic analyses were performed using HBV replication-competent plasmids carrying unique mutations selected under TDF therapy.

Results

Mean exposure to LAM was 35.3 ±27.5 months and to TDF 11.2 ±6.7 months. Genotypic analyses from 21 of the patients revealed LAM-associated mutations, and a further two patients developed a novel mutation, rtA194T, along with LAM-resistance-associated mutations. Phenotypic analyses revealed that constructs harbouring rtA194T combined with rtL180M and rtM204V displayed an over 10-fold increase in the IC50 for TDF compared with the wild type.

Conclusion

The selection of HBV mutations in HBV/HIV-coinfected patients failing TDF therapy is an unlikely event within the first 12 months of treatment. However, HBV from two of the 43 patients treated with TDF for more than 12 months was found to contain one novel mutation located distal to the catalytic site of the HBV pol. In vitro, rtA194T conferred a reduced susceptibility to TDF in the presence of LAM-associated mutations.
Design and Characterization of Human Respiratory Syncytial Virus Entry Inhibitors
Authors: Ni Ling , Zhao Linqing , Qian Yuan , Zhu Jieqing , Jin Zhibo , Chen Yu Wai , Tien Po , Gao George F
Published: April 2022

Human respiratory syncytial virus (hRSV) is a pathogen of worldwide health concern. The crucial membrane fusion event during viral entry into host cells involves a ‘trimer-of-hairpins’ structure that brings the amino (N)- and carboxy (C)-terminal regions of the viral fusion glycoprotein (F protein) into close proximity. Two heptad repeat regions that are highly conserved in the F protein - HR1 (N-terminal) and HR2 (C-terminal) - have an important role in this process. It has been shown that both HR1-and HR2-based peptides can inhibit viral entry. However, these proteins, and the HR1 peptides in particular, are liable to aggregation. We designed three peptides containing multiple copies of alternating HR1 and HR2 sequences denoted 5-Helix, HR121 and HR212, respectively. The 5-Helix, HR121 and HR212 proteins were functionally analogous to single HR1, HR1 and HR2 sequences, respectively. All three proteins were expressed in soluble form and biophysical analysis showed that they exhibited α-helical secondary structures. The three proteins were potent fusion inhibitors in vitro, at the micromolar scale, with the HR1 analogues being approximately two times more effective than the HR2 analogue. Our results suggest that these rationally designed protein inhibitors could serve as a new class of anti-hRSV agents.
Efficacy and Tolerability of the Oral Neuraminidase Inhibitor Peramivir in Experimental Human Influenza: Randomized, Controlled Trials for Prophylaxis and Treatment
Authors: Barroso Luis , Treanor John , Gubareva Larisa , Hayden Frederick G
Published: April 2022

Objective

Oseltamivir is the only oral neuraminidase inhibitor currently available; we determined the tolerability and antiviral efficacy of oral peramivir for treatment and prophylaxis of experimental human influenza A and B.

Participants

288 susceptible, healthy volunteers (ages 18–45) were inoculated intranasally with A/Texas/36/ 91/H1N1 or B/Yamagata/16/88 virus in four randomized, double-blind, placebo-controlled trials. Interventions: For treatment dosing was initiated at 24 h after inoculation with peramivir doses ranging from 100–800 mg/day for 5 days. For prophylaxis dosing was initiated 24 h before inoculation and continued for 4 days with peramivir doses ranging from 50–800 mg/day.

Outcomes

The primary outcome measure for treatment was quantitative viral detection defined by the area under the curve (AUC) for nasal wash viral titres. For prophylaxis the primary outcome measure was the incidence of virus recovery.

Results

In influenza A treatment, peramivir 400 mg q24h and 200mg q12h, but not lower doses, resulted in significant reductions in viral titre AUC. In influenza B treatment, both 400 and 800/400 mg once daily dose groups reduced AUC values. In influenza A prophylaxis, the percentage of individuals with nasal viral shedding did not differ significantly in the placebo (58%), 50 mg (61%), 200 mg (37%) and 400 mg (31%) dose groups. In influenza B prophylaxis, shedding frequencies were similar in placebo (55%), 200 mg (41%), 400 mg (35%) and 800 mg (47%) dose groups. The drug was well tolerated in all four studies, with nausea and headache being the most common side effects. No drug-resistant variants were detected.

Conclusion

Early treatment with peramivir was associated with significant antiviral effects in experimentally induced influenza in humans. Prophylaxis did not significantly reduce viral shedding. The relatively low blood peramivir concentrations observed may explain the lack of more robust antiviral effects, and parenteral dosing should be studied.
Methodological Considerations in Human Studies of Gene Expression in HIV-Associated Lipodystrophy
Authors: Mallon Patrick WG , Sedwell Rebecca , Unemori Patrick , Kelleher Anthony , Cooper David A , Carr Andrew
Published: April 2022

In the majority of cases, HIV-associated lipodystrophy, lipoatrophy in particular, becomes clinically apparent only after months or years of continuous exposure to antiretroviral medications and, once developed, is difficult to reverse. Many lipid-related side effects of antiretroviral medications result from drug-induced changes in gene expression. As our understanding of the pathogenic mechanisms underlying HIV-associated lipodystrophy improves, it is important to be able to explore changes at a molecular level in order to fully elucidate the mechanisms whereby antiretroviral drugs exert their toxicities. Monitoring changes in gene expression in vivo may enable physicians to identify, predict or prevent drug toxicities early, before irreversible changes in body composition occur. However, monitoring changes in gene expression at a population level presents many methodological challenges that need to be addressed, over and above the considerable intra- and inter-individual variability inherent in the cellular expression of any gene. Careful collection and processing of adequate biological samples, robust laboratory processes and assays, and appropriate study design can help overcome many of these difficulties.
Uridine Abrogates the Adverse Effects of Antiretroviral Pyrimidine Analogues on Adipose Cell Functions
Authors: Walker Ulrich A , Auclair Martine , Lebrecht Dirk , Kornprobst Michel , Capeau Jacqueline , Caron Martine
Published: April 2022

Objectives

Side effects of antiretroviral treatment such as lipoatrophy have been mainly attributed to mitochondrial toxicity of nucleoside reverse transcriptase inhibitors (NRTIs). We assessed whether uridine can abrogate the adverse effects of NRTIs on adipocyte functions.

Methods

3T3-F442A preadipocytes were exposed to stavudine (d4T; 10 μM), zidovudine (ZDV; 1 μM), zalcitabine (ddC; 0.2 μM) or didanosine (ddI; 10 μM) in the absence or presence of uridine 21 days prior to and 7 days after induction of differentiation. Then, lipid accumulation (oil red staining), apoptosis (flow cytometry, PARP-cleavage), mitochondrial mass (Mitotracker) and DNA (mtDNA), cytochrome c oxidase (COX) subunits and mitochondrial membrane potential (JC-1) were quantified.

Results

Whereas ddI had no effects, d4T, ZDV and ddC significantly decreased cellular lipid accumulation (by 32%, 46% and 24%, respectively), increased apoptosis and induced mitochondrial depolarization. d4T, ZDV and ddC decreased adipocyte mtDNA (by 64%, 53% and 46%, respectively) and reduced the mtDNA encoded COX II subunit. Uridine (200 μM) had no intrinsic effect, but prevented all adverse effects of d4T, ZDV and ddC on adipocyte morphology, lipid staining, apoptosis, mtDNA depletion (partial prevention with ZDV), mitochondrial mass and membrane potential. The effects of uridine were concentration-dependent. Uridine also fully reverted established d4T toxicities despite continued d4T exposure.

Conclusions

Uridine supplementation protects adipocytes from the adverse effects of d4T, ZDV and ddC on lipid accumulation, cell survival and mitochondrial functions, suggesting that the toxic effects could be linked to intracellular depletion of uridine or its metabolites. Uridine is an interesting candidate in the prevention of NRTI-induced lipoatrophy in vivo.
Novel Approaches to New Therapies for Hepatitis B Virus Infection
Authors: Loomba Rohit , Liang T Jake
Published: April 2022

Hepatitis B is one of the most prevalent viral diseases in the world. It leads to chronic liver disease in 10% of infected individuals, putting them at an increased risk for liver-related morbidity and mortality from complications of cirrhosis and hepatocellular carcinoma. Despite the success of universal hepatitis B vaccination in many countries, this disease remains a major public health problem, resulting in more than 500,000 deaths per year. Although the current therapy for chronic hepatitis B (CHB) is effective, it is not optimal; novel approaches to the management of CHB are needed. An improved understanding of virus-host interactions, advances in gene therapy, the development of molecular therapies targeted at different stages of the hepatitis B virus life cycle, and new insights into various approaches of immune modulation will lead to the development of better therapeutic agents for the management of CHB. These advances herald a new era of combination therapy. In this review, we will discuss emerging therapies and potential mechanisms, and highlight the promises and pitfalls of these new treatment strategies.
Salvage Therapy with Atazanavir/Ritonavir Combined to Tenofovir in HIV-Infected Patients with Multiple Treatment Failures: Randomized Anrs 107 Trial
Authors: Piketty Christophe , Gérard Laurence , Chazallon Corine , Marcelin Anne-Geneviève , Clavel François , Taburet Anne-Marie , Calvez Vincent , Madelaine-Chambrin Isabelle , Molina Jean-Michel , Aboulker Jean-Pierre , Girard Pierre-Marie
Published: April 2022

Background

Ritonavir (RTV)-boosted atazanavir (ATV) and tenofovir disoproxil fumarate (TDF-DF) are promising in highly experienced patients because of their pharmacokinetic profile, activity, safety and resistance properties.

Methods

A 26-week study of the safety and efficacy of RTV-boosted ATV plus TDF-DF was conducted in 53 HIV-infected patients who were failing their current highly active antiretroviral therapy (HAART) regimen. Patients with history of failure to at least two protease inhibitors (PIs) and one non-nucleoside reverse transcriptase inhibitor (NNRTI) were randomized to either continue their current regimen (group 1) or replace the PI by ATV (300 mg once daily) boosted by RTV (100 mg; group 2) for 2 weeks. Then, all patients received the same combination of ATV, RTV and TDF-DF (300 mg) plus optimized NRTIs regimen.

Results

At baseline, median CD4+ T-cell count was 206/mm3, median viral load (VL) 5.0 log10/ml and median numbers of NRTI, NNRTI and PI resistance mutations were 7, 1 and 8, respectively. At week 2, median VL remained unchanged from baseline in group 2 as compared with group 1 (-0.1 vs -0.1 log10/ml). At week 26, a mild decrease in median VL from baseline of 0.2 log10/ml was observed, with 16 (31%) and 9 (17%) patients exhibiting a decrease in viral load of at least 0.5 and 1.0 log10/ml, respectively. Baseline phenotypic and genotypic resistance to ATV were the most predictive independent factors of virological response. The regimen was well tolerated.

Conclusion

In these very advanced patients failing highly HAART, the combination of boosted ATV plus TDF-DF yielded low antiretroviral activity.
The Role of Viruses in the Aetiology of Community-Acquired Pneumonia in Adults
Authors: Marcos Maria Angeles , Camps Marta , Pumarola Tomás , Martinez José Antonio , Martinez Esteban , Mensa Josep , Garcia Elisa , Peñarroja Georgina , Dambrava Póvilas , Casas Inmaculada , de Anta Maria Teresa Jiménez , Torres Antoni
Published: April 2022

Background

The role of viruses in community-acquired pneumonia may have been previously underestimated. We aimed to study the incidence and clinical characteristics of community-acquired pneumonia (CAP) due to respiratory viruses in adults adding PCR to routine conventional laboratory tests.

Methods

Consecutive adult patients diagnosed of CAP from January 2003 to March 2004 were included. Conventional tests including cultures of blood, sputum, urine antigen detection of Streptococcus pneumoniae and Legionella pneumophila, and paired serologies were routinely performed. Nasopharyngeal swabs were processed for study of respiratory viruses through antigen detection by indirect immunofluorescence assay, isolation of viruses in cell culture and detection of nucleic acids by two independent multiplex RT-PCR assays. According to the aetiology, patients were categorized in 4 groups: group 1, only virus detected; group 2, only bacteria detected; group 3, viral and bacterial; and group 4, unkown aetiology.

Results

Of 340 patients diagnosed with CAP, 198 had nasopharyngeal swabs available and were included in this study. Aetiology was established in 112 (57%) patients: group 1, n=26 (13%); group 2, n=66 (33%); group 3, n=20 (10%). The most common aetiological agent was S. neumoniae (58 patients, 29%), followed by respiratory viruses (46 patients, 23%). Forty-eight respiratory viruses were identified: influenza virus A ( n=16), respiratory syncytial virus A ( n=5), adenovirus ( n=8), parainfluenza viruses ( n=5), enteroviruses ( n=1), rhinoviruses ( n=8) and coronavirus ( n=5). There were two patients coinfected by two respiratory viruses. Serology detected 6 viruses, immunofluorescence 8, viral culture 12, and PCR 45. For the viruses that could be diagnosed with conventional methods, the sensitivity and specificity of RT-PCR was 85% and 92%, respectively. The only clinical characteristic that significantly distinguished viral from bacterial aetiology was a lower number of leukocytes ( P=0.004).

Conclusion

PCR revealed that viruses represent a common aetiology of CAP. There is an urgent need to reconsider routine laboratory tests for an adequate diagnosis of respiratory viruses, as clinical characteristics are unable to reliably distinguish viral from bacterial aetiology.
The Safety and Tolerability of Z-100 in Patients Infected with HIV-1
Authors: Borucki Michael , Holodniy Mark , Pierone Gerald , Ruane Peter , Steinhart Corklin , Williams Sally , Slom Trevor , Palella Frank , Hare C Bradley , Yoshida Yasuhisa , Li Xiao-Dong , Pollard Richard B
Published: April 2022

Z-100 is an extract of the Mycobacterium tuberculosis strain Aoyama B, which contains various polysaccharides. Aoyama B has previously been shown to induce a T helper 1-type cytokine response in various murine oncological models and has also demonstrated inhibitory activity against HIV-1 in vitro. This multicentre study primarily determined the safety of Z-100 in early HIV-1-infected patients who were treatment naive; were treatment experienced, but had elected to discontinue highly active antiretroviral therapy (HAART) 8 weeks or longer before the study; or were stable on their first or second HAART regimen for at least 12 weeks before the study. Thirty-two individuals participated in this study and self-injected either placebo, 20 μg or 40 μg Z-100 twice a week for 8 weeks. Z-100 was well tolerated and the safety profiles of the Z-100 treatment groups were not meaningfully different compared with the placebo group. Plasma levels of HIV-1 RNA were not statistically significantly different in any treatment group at the end of the treatment period. There were no statistically significant differences among the treatment groups in the change from baseline to week 8 for any of the biological endpoints including plasma levels of HIV-1 RNA; CD4+ and CD8+ T-cell counts; levels of macrophage inflammatory protein 1; soluble tumour necrosis factor receptor 1; C-reactive protein; interleukin-6; and granulocyte colony stimulating factor. Consequently, this trial demonstrates the safety of Z-100 in HIV-1 infected patients without evidence of any activity at the doses administered.
Access to HIV Prophylaxis for Survivors of Sexual Assault: The Tip of the Iceberg
Authors: Patrizia Carrieri Maria , Karim Bendiane Marc , Paul Moatti Jean , Dominique Rey
Published: April 2022

Background

In France non-occupational post-exposure prophylaxis (nPEP) is recommended in case of sexual assault and can be prescribed, after risk evaluation, in emergency or AIDS care units.

Methods

A survey was carried out on retrospective data of individuals being consulted for nPEP in southeastern France.

Results

Of the 915 consultations for sexual exposures, 94 were sexual assaults concerning adults. Most were prescribed nPEP (91.5%), but half were lost to follow-up.

Conclusions

During the study period throughout the same region, 623 survivors reported sexual assault to the police. The comparison of these figures shows that the number of victims who have no access to medical consultation for nPEP may be alarmingly high and strongly suggests the urgency to develop strategies for guaranteeing prompt HIV medical assessment for victims of sexual assault and assure adherence to medical follow-up.
Results of a Study of Prolonging Treatment with Pegylated Interferon-α2A plus Ribavirin in HIV/HCV-Coinfected Patients with No Early Virological Response
Authors: Fuster Daniel , Planas Ramon , Gonzalez Juan , Force Lluís , Cervantes Manel , Vilaró Josep , Roget Mercè , García Isabel , Pedrol Enric , Tor Jordi , Ballesteros Angel L , Salas Anna , Sirera Guillem , Videla Sebastià , Clotet Bonaventura , Tural Cristina
Published: April 2022

Objective

To assess the efficacy and safety of an extended treatment period in HIV/hepatitis C virus (HCV)-coinfected patients without early virological response (EVR).

Methods

Patients received pegylated interferon (peg-INF)-α2a 180 μg/week plus ribavirin 800 mg/d for 12 weeks. Patients achieving EVR at week 12 continued under therapy for an additional 12 or 36 weeks depending on genotype. Patients without EVR were randomized to complete the standard treatment or treatment lasting 72 weeks (extension arm).

Results

One hundred and ten patients were included (mean age 38.7 years, mean weight 68 kg, 74% males, 74% on highly active antiretroviral therapy, mean CD4+ T-cell count 564 cells/mm3). Fifty-one patients harboured genotype 1, 44 genotype 2/3, and 15 genotype 4. Fifty-three had an HCV load >800,000 IU/ml. Premature interruptions occurred in 32.7%. EVR was achieved in 63.6% (51% in genotype 1, 88.6% in genotype 2/3, 33.3% in genotype 4). End-of-treatment response was 52.7% (47.2% in genotype 1, 68.2% in genotype 2/3, 26.7% in genotype 4). Sustained virological response (SVR) was achieved in 41.8% (37.3% in genotype 1, 54.6% in genotype 2/3, 20% in genotype 4). Only one patient allocated to the extended arm achieved SVR. The rate of drop-outs in the extension arm was 68%. The negative predictive value of EVR was 97.5%.

Conclusions

This study shows no benefit of extending therapy in patients without EVR at week 12. Measures to improve adherence to HCV antiviral therapy should be considered when new approaches based on extended periods of treatment are investigated.
Investigation of Baseline Susceptibility to Protease Inhibitors in HIV-1 Subtypes C, F, G and Crf02_Ag
Authors: Abecasis Ana B , Deforche Koen , Bacheler Lee T , McKenna Paula , Carvalho Ana Patrícia , Gomes Perpétua , Vandamme Anne-Mieke , Camacho Ricardo Jorge
Published: April 2022

Objective

To compare baseline susceptibility to protease inhibitors among HIV-1 isolates of subtypes C, F, G and CRF02_AG, and to identify polymorphisms that determine the differences in susceptibility.

Methods

A total of 42 samples of drug-naive patients infected with subtypes G ( n=19), CRF02_AG ( n=10), F ( n=6) and C ( n=7) were phenotyped and genotyped with the Antivirogram and the ViroSeq 2.0 genotyping system, respectively. A Bayesian network approach was used for a preliminary analysis of the collected data and the dependencies indicated by the network were statistically confirmed.

Results

CRF02_AG samples were found to be more susceptible to nelfinavir and ritonavir than other subtypes. Hypersusceptibility to these drugs was associated with the 70R polymorphism. 37D/S/T was associated with reduced susceptibility to indinavir and 89M with reduced susceptibility to lopinavir. Susceptibility to tipranavir was the lowest among the subtype F samples and the highest for subtype G samples, with samples carrying 57R being more susceptible than samples carrying 57K.

Conclusions

Our study suggests that there are baseline susceptibility differences between subtypes and these differences are due to naturally occurring polymorphisms in these subtypes. The predictive value for phenotype of these polymorphisms was even valid in subtypes where these polymorphisms are less prevalent. Taking into account such polymorphisms should improve current algorithms for interpretation of genotyping results in a subtype-independent way.
Protease Inhibitor Effects on Triglyceride Synthesis and Adipokine Secretion in Human Omental and Subcutaneous Adipose Tissue
Authors: Cianflone Katherine , Zakarian Robert , Stanculescu Cristina , Germinario Ralph
Published: April 2022

Objective

Significant advances in the treatment of the morbidity and mortality associated with AIDS are also associated with undesirable side-effects in fat redistribution (lipodystrophy), insulin resistance and cardiovascular risk, which is directly linked to protease inhibitor (PI) treatment.

Methods

The effects of four different PIs on triglyceride (TG) storage and adipokine production (leptin, adiponectin, and acylation stimulating protein [ASP]) in omental (OM) and subcutaneous (SC) adipose tissues were examined.

Results

Initial results demonstrated that saquinivir (SQV) and ritonivir (RTV) had little observed effect on de novo TG synthesis ([3H]glucose incorporation into TG) or fatty acid re-esterification ([14C]oleate incorporation into TG), whereas amprenivir (APV) and indinivir (IDV) reduced TG synthesis, especially in SC tissue up to 30 ±5.8% P<0.05 and 46 ±7.8% P<0.001, at 20 μM, respectively. There was no observed effect on phospholipid synthesis, tissue protein or toxicity. Only APV and IDV decreased leptin and adiponectin secretion in SC tissue, in a time- and concentration-dependent manner: at 18 h, leptin was inhibited by 54 ±3.1% ( P<0.001) and 44 ±6.4% ( P<0.001) by APV and IDV (40 μM), respectively, and adiponectin was inhibited by 35 ±5.6%( P<0.001) and 25 ±12.3% ( P<0.05) by APV and IDV (40 μM), respectively. By contrast, both IDV and APV decreased ASP secretion in OM tissues by a maximum of 53 ±7.8% ( P<0.001) and 59 ±5.9% ( P<0.001), respectively, and by a maximum of 86 ±1.6% ( P<0.001) and 72 ±4% ( P<0.001), respectively, in SC tissues.

Conclusion

PI have a direct effect on human adipose tissue which are site, PI and adipokine specific; these effects may contribute to the overall adipose imbalance and development of lipodystrophy, and metabolic syndrome in HIV-positive individuals.
Prevalence, Incidence and Predictors of Severe Anaemia with Zidovudine-Containing Regimens in African Adults with HIV Infection within the Dart Trial
Authors: Ssali Francis , Stöhr Wolfgang , Munderi Paula , Reid Andrew , Walker A Sarah , Gibb Diana M , Mugyenyi Peter , Kityo Cissy , Grosskurth Heiner , Hakim James , Byakwaga Helen , Katabira Elly , Darbyshire Janet H , Gilks Charles F
Published: April 2022

Objective

To describe the prevalence, incidence and predictors of severe anaemia in previously untreated symptomatic HIV-infected adults with CD4+ T-cells <200 cells/mm3 initiating zidovudine-containing regimens in Africa.

Design

DART is a randomized trial comparing two strategies for HIV/AIDS management in Uganda and Zimbabwe.

Methods

We analysed the occurrence of anaemia at weeks 4 and 12, and then every 12 weeks. We also evaluated sex, age, WHO stage, body mass index (BMI), baseline laboratory measurements and first regimen as predictors of developing grade 4 anaemia (<6.5 mg/dl) by week 48 using logistic regression.

Results

To May 2005, 3,314 participants (65% women, 23% at WHO stage 4, median age=37 years, baseline CD4+ T-cell=86 cells/mm3 and median baseline haemo-globin=11.4 g/dl) had a median 72 weeks follow-up. Prevalence of grade 4 anaemia was 0.7%, 2.0%, 0.5% and <0.5% at weeks 4, 12, 24 and ≥36, respectively. Overall, 219 (6.6%) participants developed grade 4 anaemia by week 48; women and those with lower haemoglobin, CD4+ T-cell count and BMI at baseline were at significantly higher risk ( P<0.05), but not those with lower neutrophils or receiving cotrimoxazole at baseline.

Conclusions

We observed a higher incidence of grade 4 anaemia than in studies from industrialized countries, which is likely to be due in part to population characteristics and in part to a higher rate of concurrent HIV-related clinical events. Clinical vigilance and haemoglobin measurements 4, 8 and 12 weeks after starting zidovudine could help to manage serious anaemia.
Antiretroviral Efficacy and Virological Profile of a Zidovudine/Lamivudine/Tenofovir Disoproxil Fumarate Combination Therapy in Antiretroviral-Naive Patients
Authors: Masquelier Bernard , Neau Didier , Boucher Sébastien , Lavignolle-Aurillac Valérie , Schrive Marie-Hélène , Recordon-Pinson Patricia , Ragnaud Jean-Marie , Fleury Hervé
Published: April 2022

Objective

To study the antiviral efficacy and the mutations selected by a triple therapy with zidovudine (AZT), lamivudine (3TC) and tenofovir disoproxil fumarate (TDF).

Methods

Antiretroviral-naive patients received 300 mg AZT/150 mg 3TC twice a day plus 300 mg TDF once a day in an open pilot study. Follow-up was assessed at baseline therapy (M0) and at months 1, 3, 6, 9 and 12. Reverse transcriptase (RT) genotypic resistance analysis and in selected cases, a recombinant drug susceptibility and replication capacity assay were performed from plasma RNA at baseline and in case of virological failure (VF); that is, rebound of viral load >50 copies/ml on therapy.

Results

Twenty-four patients were included. At baseline, the median CD4+ T-cell count was 443 cells/μl and the median plasma viral load (VL) was 4.38 log10 copies/ml. RT resistance mutations were observed at M0 in 4 patients. At M12, the proportion of patients with a VL <50 copies/ml reached 88% using an on-treatment analysis and 67% with an intent-to-treat analysis. The median increase in CD4+ T cells at M12 was 94 cells/μl. Four patients had a VF on therapy: two with wild-type viruses, one with selection of M184V and thymidine analogue mutations (TAMs) on a background of TAMs, and one with selection of K65R and M184V, with a replication capacity at 2.4%.

Conclusion

The virological response in our study demonstrates the antiviral efficacy of the AZT/3TC/TDF combination therapy, which needs further evaluation. The moderate frequency of selection of K65R could be due to the presence of AZT in the regimen.
Structurally Driven Selection of Human Hepatitis C Virus Mimotopes
Authors: Spiga Ottavia , Padula Maria G , Scarselli Maria , Ciutti Arianna , Bernini Andrea , Venditti Vincenzo , Prischi Filippo , Falciani Chiara , Lozzi Luisa , Bracci Luisa , Valensin Piero E , Caudai Cinzia , Niccolai Neri
Published: April 2022

A structural genomics approach is proposed for the development of new diagnostic kits. It combines molecular modelling, peptide synthesis and immunological tests. The preliminary step is the development of a reliable three-dimensional structure of an immunodominant protein of the target pathogenic organism using the various bioinformatic strategies that are now available to structural biologists. Once the protein structure is obtained, the most surface-exposed fragments with minimal sequence variability among the different strains reported in the genomic data bank are reproduced synthetically as linear peptides. These peptides are then tested for immunoreactivity with the plasma of infected patients to determine whether the synthetic molecules have antigenic activity and can therefore be used to detect infecting agents. This structurally driven selection of mimotopes was successfully performed for the human hepatitis C virus, as five peptides that specifically interact with the plasma of HCV-infected patients were identified solely on the basis of the three-dimensional structure predicted for the E2 homodimer of the 1a viral subtype. A similar approach could easily be extended to a large variety of immunogenic proteins from other pathogenic organisms.
Modulation of Interferon-Specific Gene Expression by Albumin-Interferon-α in Interferon-α-Experienced Patients with Chronic Hepatitis C
Authors: Balan Vijayan , Nelson David R , Sulkowski Mark S , Everson Gregory T , Lambiase Louis R , Wiesner Rusell H , Dickson Rolland C , Garcia Andy , Moore Paul A , Yu Ren , Subramanian G Mani
Published: April 2022

Albumin-interferon-α (alb-IFN) is a novel recombinant protein derived from IFN-α2b genetically fused to human albumin. The resulting single polypeptide combines in one molecule the antiviral properties of IFN-α with the long serum half-life of albumin. IFN-mediated biological responses stem from the engagement of IFN-α with its target receptor and subsequent modulation of IFN-specific gene (ISG) expression. To evaluate the pharmacodynamics of alb-IFN during the Phase I/II study conducted in patients with chronic hepatitis C (CHC) who had previously failed IFN-α-containing regimens, ISG induction was evaluated in peripheral blood and compared with antiviral response. Whole blood was obtained at day 0, day 7 and day 28 from 21 patients enrolled in the higher dose (500–900 μg) alb-IFN cohort, who received two injections on day 0 and day 14. Taqman real-time PCR was used to assess candidate ISG expression. There was sustained induction on day 7 and day 28 of the ISG's OAS1, IRF-7, IFI44 and IFI27. Although all patients showed a molecular response to alb-IFN, individual variability in pretreatment gene expression levels and fold of modulation during treatment was observed. At day 28, induction of OAS1, IFI44 and IRF7 showed pairwise correlation in individual patients ( P<0.05). Moreover, the induction of expression at day 28, and pretreatment levels of OAS1 and IFI44 correlated with hepatitis C virus RNA reduction at day 28 ( P<0.05). In conclusion, alb-IFN demonstrated robust induction of ISG that was consistent with the response associated with an IFN-α.
Antiretroviral Therapy at Conception in Pregnant Women with HIV in Italy: Wide Range of Variability and Frequent Exposure to Contraindicated Drugs
Authors:   on behalf of The Italian Group on Surveillance on Antiretroviral Treatment in Pregnancy, Floridia Marco , Tamburrini Enrica , Ravizza Marina , Anzidei Gianfranco , Tibaldi Cecilia , Bucceri Anna , Maccabruni Anna , Guaraldi Giovanni , Meloni Alessandra , Probizer Maria Fernanda Ravagni , Guerra Brunella , Martinelli Pasquale
Published: April 2022

Methods

Data from a large national surveillance study was used to describe antiretroviral regimens in pregnant women with HIV, with particular reference to the presence at conception of antiretroviral treatments contraindicated in pregnancy. Therapeutic changes during pregnancy were also analysed.

Results

Among 334 women on antiretroviral treatment at conception, less than half (42.4%) reported current pregnancy as planned. A large number of different regimens (80) was observed. All the regimens included at least one nucleoside or nucleotide reverse transcriptase inhibitor. Non-nucleoside reverse transcriptase inhibitors and protease inhibitors were present in similar proportions (39.2% and 40.7%, respectively). The most commonly used drugs were lamivudine (83.2% of regimens), zidovudine (50.0%), stavudine (d4T; 38.0%), nevirapine (25.7%), didanosine (ddI; 17.7%) and nelfinavir (17.7%). Treament with efavirenz (13.5% of regimens) and ddI+d4T (9.6%) was markedly frequent. Use of efavirenz at conception was associated with a subsequent treatment change during pregnancy (odds ratio [OR]: 13.2.; 95% confidence interval [CI]: 3.2–53.8, P<0.001). A similar but less strong association was found for ddI (OR: 1.8; 95% CI: 1.03–3.25, P=0.033), whereas being on nevirapine was associated with a lower risk (OR: 0.58; 95% CI: 0.38–0.81, P=0.013).

Conclusions

Our data show that treatment at conception frequently represents the regimen previously selected for the treatment of the non-pregnant woman. The observed rates of exposure to contraindicated treatment should lead prescribing physicians to consider in HIV-positive women therapeutic choices that take into account the likelihood of an unplanned pregnancy. Such an approach is likely to reduce not only unintended exposures to contraindicated drugs, but also therapeutic changes during pregnancy.
Virological Profiles in Hepatitis B Virus/Hepatitis C Virus Coinfected Patients under Interferon plus Ribavirin Therapy
Authors: Saitta Carlo , Pontisso Patrizia , Brunetto Maurizia R , Fargion Silvia , Gaeta Giovanni B , Niro Grazia A , Picciotto Antonio , Smedile Antonina , Squadrito Giovanni , Raimondo Giovanni
Published: April 2022

Background

Hepatitis C virus (HCV) is believed to exert a suppressive effect on hepatitis B virus (HBV) in most HBV/HCV-coinfected patients; once HCV is cured by interferon-based therapy, these patients may show HBV reactivation. However, recent evidence revealed that the virological status in HBV/HCV-untreated individuals may vary over time and may show fluctuating profiles.

Methods

To evaluate the behaviour of apparently inactive HBV infection in patients under treatment for a concurrent HCV infection, we performed a prospective study that evaluated nine consecutive patients (eight males with a median age of 45.9 years, and one female aged 62 years) longitudinally followed-up with bi-monthly evaluation of HBV/HCV viraemia levels and liver biochemistry during a 1-year treatment with interferon plus ribavirin.

Results

In seven cases the HBV infection maintained its inactive status independently of the HCV response to therapy. By contrast, two non-responder cases with persistently high HCV RNA levels showed HBV DNA flairs during the follow-up, indicating a status of active HBV infection with fluctuating virological profiles.

Conclusions

This study suggests that the HBV behaviour may be independent of the HCV activity during anti-HCV therapy in HBV/HCV-coinfected patients, and that the HBV virological profile should be monitored to recognize possible reactivations that might lead to more proper therapeutic choices or adjustments.
Exploiting Information Inherent in Binding Sites of Virus-Specific Antibodies: Design of An HCV Vaccine Candidate Cross-Reactive with Multiple Genotypes
Authors: Grollo Lara , Torresi Joseph , Drummer Heidi , Zeng Weiguang , Williamson Nicholas , Jackson David C
Published: April 2022

Background/Aims

The role of antibody in hepatitis C virus (HCV) infection remains unclear although many reports attest to its role in viral clearance. Here we describe epitopes that are recognized by antibody present in the serum of infected patients and show that such epitopes can induce neutralizing antibodies.

Methods

Human serum containing hyperimmune anti-HCV IgG was used to extract epitopes from a library of synthetic peptides that encompassed the sequences of the E1 and E2 proteins of HCV genotype 1a H77. Peptides that were bound by IgG were identified by mass spectrometry. Assembly of these epitopes with a helper T cell determinant was then carried out in order to construct candidate epitope-based vaccines.

Results

Three distinct antigenic sites were defined in the E1E2 glycoproteins by epitopes identified by antibody present in infected individuals. Four of the peptide epitopes identified are conserved in at least three HCV genotypes and are bound by antibody present in the sera of chronically infected and convalescent individuals. Synthetic vaccines based on these epitopes elicited antibodies that are capable of (i) capturing HCV virions from the serum of viraemic patients and (ii) inhibiting HCV pseudovirus particle entry into Huh7 cells.

Conclusions

This approach exploits the information inherent in the binding sites of virus-specific antibodies and represents a novel method for the design of synthetic epitope-based vaccines.
The Development of Artificial Neural Networks to Predict Virological response to Combination HIV Therapy
Authors: Larder Brendan , Wang Dechao , Revell Andrew , Montaner Julio , Harrigan Richard , De Wolf Frank , Lange Joep , Wegner Scott , Ruiz Lidia , Pérez-Elías María Jésus , Emery Sean , Gatell Jose , Monforte Antonella D‘Arminio , Torti Carlo , Zazzi Maurizio , Lane Clifford
Published: April 2022

Introduction

When used in combination, antiretroviral drugs are highly effective for suppressing HIV replication. Nevertheless, treatment failure commonly occurs and is generally associated with viral drug resistance. The choice of an alternative regimen may be guided by a drug-resistance test. However, interpretation of resistance from genotypic data poses a major challenge.

Methods

As an alternative to current interpretation systems, we have developed artificial neural network (ANN) models to predict virological response to combination therapy from HIV genotype and other clinical information.

Results

ANN models trained with genotype, baseline viral load and time to follow-up viral load (1,154 treatment change episodes from multiple clinics), produced predictions of virological response that were highly significantly correlated with actual responses (r2=0.53; P<0.00001) using independent test data from clinics that contributed training data. Augmented models, trained with the additional variables of baseline CD4+ T-cell count and four treatment history variables, were more accurate, explaining 69% of the variance in virological response. Models trained with the full input dataset, but only those data involving highly active antiretroviral therapy (three or more full-dose antiretroviral drugs in combination), performed at an intermediate level, explaining 61% of the variance. The augmented models performed less well when tested with data from unfamiliar clinics that had not contributed data to the training dataset, explaining 46% of the variance in response.

Conclusion

These data indicate that ANN models can be quite accurate predictors of virological response to HIV therapy even for patients from unfamiliar clinics. ANN models therefore warrant further development as a potential tool to aid treatment selection.
HIV-1 drug resistance surveillance using dried whole blood spots
Authors: Bertagnolio Silvia , Soto-Ramirez Luis , Pilon Richard , Rodriguez Roberto , Viveros Monica , Fuentes Luis , Harrigan P Richard , Mo Theresa , Sutherland Don , Sandstrom Paul
Published: April 2022

Background

Field-friendly methods for HIV drug resistance (HIVDR) surveillance in resource-limited regions are urgently needed. Despite evidence that dried blood spots (DBS) are suitable for HIV serology, viral load and CD4+ T-cell enumeration, no study has evaluated DBS for HIVDR genotyping. We assessed the feasibility of genotyping HIV-1 from field-collected DBS stored under challenging environmental conditions.

Methods

We prospectively collected specimens from newly diagnosed, treatment-naive HIV-positive subjects in Mexico. Whole blood was spotted onto filter cards, air dried at ambient temperature and stored with desiccant at 37°C and 85% humidity for 3 months. Genotypes obtained from DBS-extracted nucleic acids using an in-house nested reverse transcription-PCR method were compared to genotypes derived from matched plasma.

Results

Genotypes from 103 phylogenetically matched plasma and DBS were compared. In total, 90.1% of all DBS specimens could be amplified in either the region of HIV protease or the region of reverse transcriptase. Failure to amplify from DBS did not correlate with low plasma viral loads. Between paired specimens, the median nucleotide similarity was 99.95%. In the nine specimens with drug resistance mutations, all differences between pairs were partial discordances. Mutations identified in plasma were found in the majority of replicate DBS amplifications.

Conclusion

The results suggest that genotypes obtained from DBS are equivalent to those from plasma. DBS are a promising public health tool for HIVDR surveillance of treatment-naive subjects, especially in regions where specimens might be exposed to severe environmental conditions and where logistical difficulties could prevent timely specimen processing. More studies are needed to validate DBS for patient monitoring.
Assessing Theoretical Risk and Benefit suggested by Genetic Association Studies of CCR5: Experience in a Drug Development Programme for Maraviroc
Authors: Wheeler Jeremy , McHale Mary , Jackson Vicky , Penny Michelle
Published: April 2022

The proliferation of published gene association studies of the CCR5A32 mutation is of relevance to drug development of a CCR5 antagonist for HIV, in highlighting potential safety concerns. We conducted an initial review of all non-HIV gene association studies of CCR5-Δ32, followed by detailed meta-analyses in the three disease areas most commonly reported. Our review indicated no consistent evidence of increased risk of susceptibility to hepatitis C virus infection or multiple sclerosis among individuals with CCR5-Δ32 mutation, and suggested treatment with a CCR5 inhibitor is unlikely to have related adverse effects. There was, however, evidence to suggest rheumatoid arthritis as a potential therapeutic target for a CCR5 antagonist. Clinical evidence would be required to confirm these findings.
Molecular Assessment of the Potential for Renal Drug Interactions between Tenofovir and HIV Protease Inhibitors
Authors: Cihlar Tomas , Ray Adrian S , Laflamme Genevieve , Vela Jennifer E , Tong Leah , Fuller Michael D , Roy Anupma , Rhodes Gerald R
Published: April 2022

Background

Active renal secretion of tenofovir (TFV) across proximal tubules occurs via uptake by human organic anion transporters 1 and 3 (hOAT1 and hOAT3) coupled with efflux by multidrug resistance protein 4 (MRP4). Co-administration of some HIV protease inhibitors (PIs) with tenofovir disoproxil fumarate (TDF), an oral prodrug of TFV, has been shown to increase systemic levels of TFV, leading to a hypothesis that PIs may affect tubular secretion of TFV and potentially alter the renal safety of TDF.

Methods

The effect of PIs on the transport of TFV by hOAT1, hOAT3 and MRP4 was assessed using in vitro cell-based transport models.

Results

At concentrations equal to their therapeutic peak plasma levels (Cmax) all PIs showed <20% inhibition of TFV transport by hOAT1. hOAT3 was more sensitive to PIs with ritonavir (RTV) and lopinavir being the most potent inhibitors of TFV transport (62% and 37% inhibition, respectively, at their Cmax). In the absence of human serum, RTV at concentrations exceeding its therapeutic Cmax also exhibited a minor effect on the cellular efflux of TFV by MRP4 (<30% inhibition at 20 μM). However, no effects of PIs on hOAT1, hOAT3 or MRP4 were detected in the presence of human serum with the exception of RTV that inhibited hOAT3 by approximately 35% at its Cmax. In addition, PIs did not affect the cytotoxicity of TFV or TDF in MRP4- or MRP2-overexpressing cells.

Conclusion

These data indicate a low potential of PIs to interfere with the active tubular secretion of TFV and to alter the clinical renal safety profile of TDF.
Risk of Discontinuation of Nevirapine due to Toxicities in Antiretroviral-Naive and -Experienced HIV-Infected patients with High and Low CD4+ T-cell Counts
Authors: Mocroft Amanda , Staszewski Schlomo , Weber Rainer , Gatell José , Rockstroh Jurgen , Gasiorowski Jacek , Panos George , Monforte Antonella d'Arminio , Rakhmanova Aza , Phillips Andrew N , Lundgren Jens D
Published: April 2022

Introduction

It is unknown whether the increased risk of toxicities in antiretroviral-naive HIV-infected patients initiating nevirapine-based (NVPc) combination antiretroviral therapy (cART) with high CD4+ T-cell counts is also observed when NVPc is initiated in cART-experienced patients.

Patients and methods

1,571 EuroSIDA patients started NVPc after 1/1/1999, with CD4+ T-cell counts and viral load measured in the 6 months before starting treatment, and were stratified into four groups based on CD4+ T-cell counts at initiation of NVPc (high [H], >400/mm3 or >250/mm3 for male or female, respectively, or low [L], ≤400/mm3 or ≤250/mm3 for male or female) and prior antiretroviral experience (antiretroviral-naive [N] or -experienced [E]). Cox proportional hazards models compared the risks of discontinuation of nevirapine due to toxicities or patient/physician choice (TOXPC).

Results

After adjustment, there was a significantly lower risk of discontinuation of nevirapine due to TOXPC in the HE group ( n=588; proportion discontinued by 3/12 months: 10/17%, respectively) than in HN (n=62; 21/32% respectively; overall relative hazard [RH]: 0.56; 95% confidence interval [CI]: 0.34–0.94; P=0.027). This difference was most pronounced during the first 3 months of NVPc (RH: 0.44; 95% CI: 0.23–0.87; P=0.017). There were no deaths in the 6 months after starting NVPc resulting from exposure to <3 months of NVPc exposure within the HE group (incidence: 0; per 1,000 person-years follow up; 95% CI: 0–6.9). After adjustment, there were no differences between the HE and HN groups in discontinuation due to TOXPC in patients starting efavirenz-based cART (RH: 0.91; 95% CI: 0.60–1.38; P=0.66) or protease-inhibitor-based cART (RH: 1.13; 95% CI: 0.77–1.66; P=0.52).

Conclusions

Results from this non-randomized study suggest that NVPc might be safer to initiate in antiretro-viral-experienced than in antiretroviral-naive patients with high CD4+T-cell counts.