bluuebunny/crossref_metadata_2025 huggingface dataset:
Activation of Dendritic Cells and Induction of T Cell Responses by Hpv 16 L1/E7 Chimeric Virus-Like Particles are Enhanced by Cpg ODN or Sorbitol
Authors: Freyschmidt Eva-Jasmin , Alonso Angel , Hartmann Gunther , Gissmann Lutz
Published: April 2022

Chimeric human papillomavirus-like particles, consisting of human papillomavirus (HPV) 16 L1-E7 fusion proteins [HPV 16 L1/E7 chimeric virus-like particles (CVLP)], are a vaccine candidate for treatment and prevention of cervical cancer. Although in preclinical studies CVLPs were shown to induce neutralizing antibodies and L1-and E7-specific T cell responses, the results of a recent clinical trial emphasized the need of improved immunogenicity of CVLPs. Here we studied the interaction of HPV 16 L1/E7 CVLPs with mouse bone marrow-derived dendritic cells (BMDCs) activated with different immune adjuvants. We found that lipopolysaccharides (LPS), unmethylated CpG motifs (CpG ODN) and sorbitol enhanced CVLP-induced stimulation of C57BL/6 mouse BMDCs as revealed by increased levels of CD40, CD80, MHC II and CD54 at the cell surface. CpG ODN and sorbitol also enhanced the presentation of Db-restricted cytotoxic T lymphocyte epitopes to HPV 16 L1- or E7-specific T lymphocytes after loading of CVLPs onto BMDCs. Treatment of BMDCs with CpG ODN in combination with CVLPs improved in vitro priming of naive T lymphocytes by CVLP-loaded BMDCs. In vivo, CVLP-loaded BMDCs were more immunogenic as compared with injection of CVLPs alone. CpG ODN and sorbitol further enhanced priming of antigen-specific T cell responses. Our data demonstrate that CpG ODN- or sorbitol-activated BMDCs substantially increase the immunogenicity of CVLPs. Implementing our results in clinical trial protocols may lead to improved activity of therapeutic HPV vaccines for the treatment of HPV-induced cancer.
The Management of Hepatitis B Virus/HIV-1 Co-Infected Patients Starting Their First Haart Regimen. Treating Two Infections for the Price of One Drug?
Authors:   the HepaI.CoNA,   the ICoNA Study Group, Puoti Massimo , Cozzi-Lepri Alessandro , Ancarani Fausto , Bruno Raffaele , Ambu Silvia , Ferraro Teresa , Tundo Paolo , Santantonio Teresa , Toti Mario , Bonasso Marino , Monforte Antonella d'Arminio , Ancarani F , Antonucci G , Bonasso M , Bruno R , Cozzi-Lepri A , Monforte A d'Arminio , Luca A De , Galli M , Gennero L , Girardi E , Lipani F , Marino N , Milazzo L , Morsica G , Narciso P , Pizzaferri P , Puoti M , Santantonio T , Verucchi G , Montroni M , Scalise G , Zoli A , Prete MS Del , Tirelli U , Di Gennaro G , Pastore G , Ladisa N , Minafra G , Suter F , Arici C , Chiodo F , Colangeli V , Fiorini C , Coronado O , Carosi G , Cadeo GP , Castelli F , Minardi C , Vangi D , Rizzardini G , Migliorino G , Manconi PE , Piano P , Ferraro T , Scerbo A , Pizzigallo E , D'Alessandro M , Santoro D , Pusterla L , Carnevale G , Galloni D , Viganò P , Mena M , Ghinelli F , Sighinolfi L , Leoncini F , Mazzotta F , Pozzi M , Lo Caputo S , Angarano G , Grisorio B , Ferrara S , Grima P , Tundo P , Pagano G , Piersantelli N , Alessandrini A , Piscopo R , Toti M , Chigiotti S , Soscia F , Tacconi L , Orani A , Perini P , Scasso A , Vincenti A , Chiodera F , Castelli P , Scalzini A , Fibbia G , Moroni M , Lazzarin A , Cargnel A , Vigevani GM , Caggese L , d'Arminio Monforte A , Repetto D , Novati R , Galli A , Merli S , Pastecchia C , Moioli MC , Esposito R , Mussini C , Abrescia N , Chirianni A , Izzo C , Piazza M , De Marco M , Montesarchio V , Manzillo E , Graf M , Colomba A , Abbadessa V , Prestileo T , Mancuso S , Ferrari C , Pizzaferri P , Filice G , Minoli L , Bruno R , Novati S , Balzelli F , Loso K , Petrelli E , Cioppi A , Alberici F , Ruggieri A , Menichetti F , Martinelli C , De Stefano C , La Gala A , Ballardini G , Briganti E , Magnani G , Ursitti MA , Arlotti M , Ortolani P , Ortona L , Dianzani F , Ippolito G , Antinori A , Antonucci G , D'Elia S , Narciso P , Petrosillo N , Vullo V , De Luca A , Del Forno L , Zaccarelli M , Acinapura R , De Longis P , Ciardi M , D'Offizi G , Trotta MP , Noto P , Lichtner M , Capobianchi MR , Girardi E , Pezzotti P , Rezza G , Mura MS , Mannazzu M , Caramello P , Sinicco A , Soranzo ML , Gennero L , Sciandra M , Bonasso M , Grossi PA , Basilico C , Poggio A , Bottari G , Raise E , Pasquinucci S , De Lalla F , Tositti G , Resta F , Chimienti A , Cozzi-Lepri A
Published: April 2022

We examined the impact of a lamivudine-containing highly active antiretroviral therapy (HAART) regimen on 164 hepatitis B virus/HIV co-infected individuals starting their first HAART. Lamivudine-treated patients (accounting for 73% of the study population) showed a significantly lower level of alanine aminotransferase over follow-up [–81.1 mU/ml mean difference; 95% confidence intervals (95% CI): –30.3; –131.7, P=0.003] and a significantly reduced risk of liver-related morbidity/mortality [Relative hazard (RH)=0.07; 95% CI: 0.01–0.38, P=0.002] than those starting a lamivudine sparing-regimen.
Hpv Proteins as Targets for Therapeutic Intervention
Authors: Grm Helena Sterlinko , Banks Lawrence
Published: April 2022

Human papillomaviruses (HPV) are the aetiological agents of several types of anogenital tumours, particularly cervical carcinoma. Recent evidence also suggests a role for HPV in the development of squamous cell carcinomas of the skin, especially in immunocompromised individuals. HPV infection also produces a number of non-malignant, but nonetheless cosmetically unpleasant lesions. Therefore, any effective therapeutic treatment for HPV-induced diseases would be extremely beneficial both on humanitarian grounds as well as being economically very attractive. In this review, we will discuss the functions of the viral proteins that appear to be the most appropriate for the development of therapeutics aimed at the treatment of viral infection and virus-induced cancers.
Alternation of Antiretroviral Drug Regimens for HIV Infection. Efficacy, Safety and Tolerability at Week 96 of the Swatch Study
Authors:   on behalf of the Swatch Study Team, Negredo Eugenia , Paredes Roger , Peraire Joaquim , Pedrol Enric , Côté Helene , Gel Silvia , Fumaz Carmina R , Ruiz Lidia , Abril Vicente , de Castro Eduardo Rodriguez , Ochoa Claudia , Martinez-Picado Javier , Montaner Julio , Rey-Joly Celestino , Clotet Bonaventura , Clotet B , Ruiz L , Martinez-Picado J , Gel S , Fumaz CR , Muñoz-Moreno JA , Bonjoch A , Martínez JC , Miranda J , Puig J , Arisa ER , Tuldrà A , Bonjoch A , Jou A , Tural C , Sirera G , Romeu J , Negredo E , Zala C , Ochoa C , Cahn Pedro , Torres O , Domingo P , Vilaró J , Llibre JM , Peraire J , Vidal F , Richart C , Viladés C , Martín L , Rodríguez R , Mata R , Viciana P , Abril V , Rubio R , Torralba M , Cervantes M , Gatell JM , Lonca M , Ruiz I , Azuaje C , Pedrol E , Rodríguez de Castro E
Published: April 2022

Introduction

Alternation of antiretroviral drug regimens has been proposed as a novel treatment strategy for HIV infection. However, some concerns persist regarding antiviral efficacy, adherence, toxicity and resistance evolution in the long term.

Methods

A total of 161 antiretroviral-naive HIV-1-infected patients were randomized to receive stavudine/ didanosine/efavirenz (group A) or zidovudine/lamivudine/ nelfinavir (group B) or to alternate between the two regimens every 3 months starting with regimen A (group C). Antiviral efficacy, adherence, safety and tolerability were analysed every 12 weeks.

Results

After 96 weeks, time to virological failure was significantly delayed in the alternating regimen compared with the standards of care regimens. Virological suppression was seen in 46%, 48% and 58% of patients in groups A, B and C, respectively, in the intention-to-treat analysis and in 75%, 76% and 97% in the on-treatment analysis (A vs C: P=0.014; B vs C: P=0.016; A vs B: P=0.849). At the end of the study, 94% of patients in group A and 92% in groups B and C reported an adherence greater than 95%. Alternating therapy was associated with a similar impact on CD4+ counts in comparison with the standards of care regimens, as well as a lower mitochondrial DNA/nuclear DNA (mtDNA/nDNA) ratio decrease in the mitochondrial substudy performed on 37 patients. The frequency and intensity of adverse events in the alternating group decreased during subsequent cycles.

Discussion

Our results favour the hypothesis that proactive therapy switching may delay the accumulation of resistance mutations. Moreover, the alternating regimen was well tolerated and adherence remained comparably high in all treatment groups. The lower mtDNA/nDNA ratio decrease observed in this group may imply a lower impact on mitochondrial toxicity than in standard regimens.
Depressive Symptoms after Initiation of Interferon Therapy in Human Immunodeficiency Virus-Infected Patients with Chronic Hepatitis C
Authors: Laguno Montserrat , Blanch Jordi , Murillas Javier , Blanco Jose Luis , León Agathe , Lonca Montserrat , Larrousse Maria , Biglia Alejandra , Martinez Esteban , García Felipe , Miró Jose M , de Pablo Joan , Gatell Josep M , Mallolas Josep
Published: April 2022

Background

Interferon alpha (IFN-α) therapy for chronic hepatitis C (CHC) infection is commonly associated with neuropsychiatric side effects including depressive symptomatology. In this study, we evaluated the incidence and management of depressive symptoms during IFN-α therapy in HIV-infected patients with CHC.

Methods

HIV-infected patients with CHC who began IFN-α and ribavirin therapy during the recruitment period April 2001 to April 2003 were included in the study. Patients with a history of major depressive disorder were excluded.

Results

Of 113 co-infected patients who started IFN-α therapy during the recruitment period, 45 (40%) developed symptoms of depression (sadness, tiredness and apathy). Twenty of them (44%) were treated with citalopram, a selective serotonin re-uptake inhibitor, resulting in a significant improvement in their symptoms. Most of the patients (60%) showed depressive side effects in the first 3 months after initiation of IFN-α. In addition, during the study, three patients developed psychotic symptoms and one committed suicide.

Conclusions

The incidence of depressive symptoms in patients with HIV/HCV co-infection treated with IFN-α is high. Most of the depressive symptoms were not severe and improved with antidepressant therapy, without reduction or cessation of IFN-α therapy. During the first weeks after initiating IFN-α therapy for HIV/HCV co-infection, close assessment of psychiatric symptoms is recommended. Early treatment of these side effects with antidepressants would help avoid early dropouts from interferon therapy.
A Randomized Controlled Trial to Enhance Antiretroviral Therapy Adherence in Patients with a History of Alcohol Problems
Authors: Samet Jeffrey H , Horton Nicholas J , Meli Seville , Dukes Kim , Tripp Tara , Sullivan Lisa , Freedberg Kenneth A
Published: April 2022

Objective

To assess the effectiveness of an individualized multicomponent intervention to promote adherence to antiretroviral therapy (ART) in a cohort of HIV-infected individuals with a history of alcohol problems.

Design

We conducted a randomized controlled trial to compare the usual medical follow-up with an adherence intervention.

Setting

The principal enrolment site was Boston Medical Center, a private, not-for-profit, academic medical institution.

Subjects

HIV-infected patients with a history of alcohol problems on ART. A total of 151 were enrolled and 141 (93%) were assessed at follow-up.

Intervention

A nurse, trained in motivational interviewing, completed the following over 3 months in four encounters: addressed alcohol problems; provided a watch with a programmable timer to facilitate pill taking; enhanced perception of treatment efficacy; and delivered individually tailored assistance to facilitate medication use.

Main outcome measures

Prior 30-day adherence ≥95%, prior 3-day adherence of 100%, CD4 cell count, HIV RNA and alcohol consumption, each at both short- and long-term follow-up.

Results

At follow-up, no significant differences in medication adherence, CD4 cell count, HIV RNA or alcohol consumption were found (all P values >0.25). Conclusions: A multicomponent intervention to enhance adherence among HIV-infected individuals with a history of alcohol problems was not associated with changes in medication adherence, alcohol consumption or markers of HIV disease progression. The failure to change adherence in a group at high risk for poor adherence, despite utilizing an intensive individual-focused patient intervention, supports the idea of addressing medication adherence with supervised medication delivery or markedly simplified dosing regimens.
Similar Reduction of Cytomegalovirus DNA Load by Oral Valganciclovir and Intravenous Ganciclovir on Pre-Emptive Therapy after Renal and Renal–Pancreas Transplantation
Authors: Kalpoe Jayant S , Schippers Emile F , Eling Yoav , Sijpkens Yvo W , de Fijter Johan W , Kroes Aloys CM
Published: April 2022

Background

Pre-emptive treatment of CMV infection in transplant recipients aims at prevention of clinical disease by early detection. However, current treatment requires the intravenous (iv) administration of ganciclovir for 2 weeks, which is a considerable burden for the patient. In this observational study, the efficacy of the new oral prodrug valganciclovir was compared with iv ganciclovir.

Methods

To facilitate the introduction of valganciclovir, a therapeutic guideline was developed to use this drug under controlled conditions with regard to safety in renal/renal–pancreas transplant recipients requiring CMV therapy. Subsequently, a group of 57 consecutive transplant recipients was evaluated. Onset and treatment of CMV infections were followed by frequent monitoring of CMV DNA in plasma by quantitative real-time PCR. Details of antiviral therapy were documented.

Results

In 15 out of 57 transplant recipients, a total of 27 anti-CMV treatment episodes were recorded: 18 with valganciclovir (900 mg twice daily) and nine with iv ganciclovir (5 mg/kg twice daily) as initial treatment. Median CMV DNA load reduction during treatment was 0.12 log10/day in the valganciclovir group and 0.09 log10/day in the ganciclovir group. There were no haematological side effects in any group and no patient developed signs of clinical CMV disease.

Conclusion

Similar reduction of CMV DNA load was observed during pre-emptive treatment with oral valganciclovir and iv ganciclovir in transplant recipients. Oral valganciclovir would provide an attractive and safe alternative for pre-emptive CMV treatment in renal/renal–pancreas transplant patients, however, confirmation in larger randomized studies would be desirable.
Influence of Rosiglitazone on Flow-Mediated Dilation and Other Markers of Cardiovascular Risk in HIV-Infected Patients with Lipoatrophy
Authors: Kovacic Jason C , Martin Allison , Carey Dianne , Wand Handan , Mallon Patrick WG , Feneley Michael P , Emery Sean , Cooper David A , Carr Andrew
Published: April 2022

Background

Antiretroviral therapy for HIV infection is commonly complicated by lipoatrophy, insulin resistance and dyslipidaemia. In HIV-uninfected adults with insulin resistance or type 2 diabetes, thiazolidinediones can lower blood pressure and improve both insulin sensitivity and endothelial function. This study sought to investigate the effects of rosiglitazone on endothelial function and other markers of cardiovascular risk in patients with HIV-related lipoatrophy.

Methods

HIV-infected, lipoatrophic adults receiving anti-retroviral therapy were randomized to receive either rosiglitazone 4 mg or matched placebo, twice daily. Percentage flow-mediated forearm arterial dilation (FMD%) was measured at weeks 0, 12, 24 and 48, together with other markers of vascular risk (blood pressure, lipids, glycaemic parameters, adiponectin and leptin).

Results

Out of 64 enrolled adults, 44 (69%) attended all visits (23 rosiglitazone, 21 placebo). Relative to placebo, at week 48, rosiglitazone decreased systolic blood pressure (8 mmHg, P=0.03), insulin (3 μIU/ml, P=0.02), insulin resistance ( P=0.03) and leptin (0.6 ng/ml, P=0.02), whilst adiponectin was increased (3.3 μg/ml, P<0.0001). However, rosiglitazone increased total cholesterol (49.1 mg/dl, P=0.001), low-density lipoprotein cholesterol (23.5 mg/dl, P=0.01) and triglycerides (146 mg/dl, P=0.06). Mean baseline FMD% for the entire cohort was moderately impaired (4.5%). Compared with baseline, mean on-treatment FMD% increased by 0.8% with rosiglitazone and decreased by 0.3% with placebo (mean difference 1.1%, 95% CI -0.2 to 2.5, P=0.09).

Conclusions

Rosiglitazone has minimal effect on flow-mediated dilation in HIV-infected lipoatrophic adults. However, despite worsening of the lipid profile, the overall effect of rosiglitazone on the cardiovascular risk profile in these subjects was positive.
A Study to Investigate the Impact of the Initiation of Highly Active Antiretroviral Therapy on the Hepatitis C Virus Viral Load in HIV/HCV-Coinfected Patients
Authors: Mijch Anne , Sasadeusz Joseph , Hellard Margaret , Dunne Mandy , McCaw Rhonda , Bowden Scott , Gowans Eric J
Published: April 2022

Changes in the hepatitis C virus (HCV) viral load (VL) were assessed in a retrospective study of 50 HIV/HCV-coinfected patients who initiated highly active antiretroviral therapy (HAART). Most patients responded to HAART [during the first 6 months, plasma HIV VL fell by a mean 1.39 log10, becoming undetectable (<400 copies/ml) in 22% and CD4+ T cells increased by a mean of 100 cells/μl], but surprisingly, 27 (54%) showed some rise and 25 (50%) showed a significant increase in the HCV VL. This figure was considered to be a minimum estimate. A majority of the patients showed an increase of less than 1 log10 that was associated with a rapid decrease in the HIV VL, whereas an increase in the HCV VL of greater than 1 log10, noted in eight patients, was associated with a baseline CD4+ cell count of less than 200 cells/μl. The increase in the HCV VL was not associated with hepatitis as determined by raised alanine transferase.
Mitochondrial Toxicity of Nucleoside Analogues in Primary Human Lymphocytes
Authors: Setzer Bernhard , Schlesier Michael , Thomas Anna K , Walker Ulrich A
Published: April 2022

Objective

To evaluate if nucleoside analogue reverse transcriptase inhibitors (NRTIs) and polymerase-gamma inhibitors deplete mitochondrial DNA (mtDNA) in cultured primary lymphocytes and if such depletion might be associated with functional defects.

Methods

Primary peripheral blood CD4 and CD8 lymphocytes were purified from six healthy humans (three male and three female), stimulated mitotically (CD3/CD28) and cultured for 10 days in the presence or absence of NRTIs. Lymphocyte proliferation, mtDNA content, the expression of mtDNA-encoded cytochrome c-oxidase II (COXII) and lactate production were assessed.

Results

In CD4 lymphocytes, 10-day exposure to zalcitabine (1.77 μM), didanosine (118 μM) and stavudine (36 μM) induced a time-dependent decline of mtDNA. Compared with controls, residual mtDNA levels were 25%, 21% and 40%, respectively. COXII was reduced to 55%, 35% and 70% of control values. Lactic acid production was increased (by 214%, 294% and 175%, respectively). At day 10, lymphocyte counts were reduced (to 60%, 51%, and 41%, respectively). Zidovudine (71 μM) also reduced lymphocyte counts to 34% and increased lactic acid production by 170%, but did not induce mtDNA and COXII depletion. All these changes were highly significant. Lower NRTI concentrations (0.177 μM of zalcitabine, 11.8 μM of didanosine, 3.6 μM of stavudine and 7.1 μM of zidovudine) had effects at the border of significance. Similar observations were made in CD8 lymphocytes.

Conclusions

In human lymphocytes, zalcitabine, didanosine and stavudine induce dose- and time-dependent mtDNA depletion, which is associated with decreased cell proliferation and increased lactate production. Zidovudine impairs lymphocyte division without inducing mtDNA depletion.
Clinical Characteristics of Iris Syndrome in Patients with HIV and Tuberculosis
Authors: Michailidis Christos , Pozniak Anton L , Mandalia Sundhiya , Basnayake Sheena , Nelson Mark R , Gazzard Brian G
Published: April 2022

Background

Some patients with HIV/tuberculosis (TB) coinfection who are on anti-TB treatment and highly active antiretroviral therapy (HAART) will develop an exacerbation of symptoms, signs or radiological manifestations of TB that are not due to relapse or recurrence of their TB. The aetiology of these immune reconstitution inflammatory syndrome (IRIS) reactions is unknown but it is presumed that they occur, at least in part, as a consequence of HAART-related reconstitution of immunity.

Methods

Patients who were diagnosed with their first episode of definitive or presumed TB between January 2001 and July 2003 were identified from the Chelsea and Westminster TB/HIV database. The patients were classified into those who developed IRIS and those who did not using a set definition of the syndrome. Demographic, clinical and laboratory data relating to both HIV and TB were compared between the two groups.

Results

A total of 55 cases of TB were identified, of which 45 cases were confirmed on culture or gene probe and 10 were presumed cases. Fourteen cases (25.5%) developed IRIS with a median (range) duration of 2.53 (0.53–14.97) months. The median baseline CD4 [interquartile range (IQR)] for the IRIS group was significantly lower at 80 (33–117) cells/mm3 ( P=0.05) than the non-IRIS group at 139 (77–284) cells/mm3. A significantly greater proportion of patients in the IRIS group [11/14 (78.6%), P=0.011] had baseline CD4 <100 cells/mm3 compared with the non-IRIS group [16/41 (39.0%)]. There was no significant difference between the two groups when comparing the log10 baseline viral load (VL). Eight (57.0%) patients in the IRIS group had disseminated TB at baseline compared with seven (17.0%) in the non-IRIS group ( P=0.006). In those who had a detectable VL at baseline, the median fold change (IQR) in CD4 from baseline to 3 months was significantly higher in the IRIS group patients, 1.5 (0.6–5.6), compared with 0.7 (-0.2 to 1.0) for those in the non-IRIS group ( P=0.046).

Conclusions

Patients who develop IRIS are more likely to present with disseminated TB, have a CD4 count <100 cells/mm3 and have a prompt rise in CD4 count in the initial 3 months of HAART.
Inhibition of Replication and Infection of Severe Acute Respiratory Syndrome-Associated Coronavirus with Plasmid-Mediated Interference RNA
Authors: Ni Bing , Shi Xinfu , Li Yang , Gao Wenda , Wang Xiliang , Wu Yuzhang
Published: April 2022

Severe acute respiratory syndrome (SARS) is a newly emerged infectious disease caused by a novel coronavirus (SARS-CoV), which spread to over 30 countries in early 2003. Until recently, no specific vaccines and effective drugs have been available to protect patients from infection by this virus. To exploit a new strategy to fight this disease, we investigated the effect of interference RNA (RNAi) on the virus infection and replication with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT), plaque-forming, Western-blot and real-time PCR assays. Results showed that the plasmid-derived siRNAs targeting the non-structural protein 1 (NSP1) sequence of the SARS-CoV genome could specifically inhibit the expression of the NSP1 sequence and effectively suppress the replication and propagation of SARS-CoV in cultured Vero E6 cell lines. The expression of the Spike and Nucleoprotein genes of SARS-CoV at mRNA and protein levels in small interfering (si)RNA-expressing cells was significantly less than that in controls when analysed with PCR and Western-blot assays, 3 days post SARS-CoV infection. Our study provides strong evidence that the NSP1 sequence in the SARS-CoV genome is a valid target for RNAi and the effect of the siRNAs probably mainly resulted from effects on global reduction of subgenome synthesis and subsequent protein expression of SARS-CoV.
The Clinical Utility of Using Catrimox-14-Treated Whole Blood in Detecting Hepatitis C Virus RNA
Authors: Patel Keyur , Albrecht Jeff , Owens Erin , Dev Anouk , Heaton Shanon , Pianko Stephen , Pockros Paul J , Conrad Andrew , Blatt Lawrence M , McHutchison John G
Published: April 2022

Background

Measuring hepatitis C virus (HCV) RNA in serum or plasma may underestimate the true HCV burden. Extracting viral RNA from whole blood (WB) with a cationic surfactant (Catrimox-14) has resulted in HCV RNA concentrations up to 1000-fold higher than from serum or plasma in some studies, but not others. We compared the Catrimox-14 WB assay with a standard serum assay.

Methods

Seventy-two chronic HCV patients received 48 weeks of standard or pegylated interferon alpha-2b and ribavirin therapy. Catrimox-14-treated WB and corresponding serum samples were obtained at baseline and weeks 12, 24, 48 and 72. HCV RNA concentrations from WB and serum were quantified by a previously validated RT-PCR assay.

Results

Overall mean (±SD) baseline serum log10 HCV RNA concentration was 6.5 ((±0.58) copies/ml. Out of 72 patients, 33 had no detectable virus at 72 weeks. Neither assay detected virus in these patients at 12 weeks and neither WB nor serum assays detected virus at end-of-treatment in the 10 patients who subsequently relapsed at 72 weeks. HCV RNA concentrations from WB and serum assays were linearly correlated (R2=0.73; P<0.001), although mean serum HCV RNA concentrations were 0.5 log10 copies/ml higher in serum than in WB [6.0 (±0.82) vs 5.5 ((±0.84), respectively, P=0.12].

Conclusions

Catrimox-14-treated WB assays detect changes in HCV RNA, but do not offer clinical advantage over a conventional serum RT-PCR based assay.
Reverse Transcriptase Inhibitors Alter Uncoupling Protein-1 and Mitochondrial Biogenesis in Brown Adipocytes
Authors: Rodríguez de la Concepción M Luisa , Yubero Pilar , Domingo Joan C , Iglesias Roser , Domingo Pere , Villarroya Francesc , Giralt Marta
Published: April 2022

Objective

Human adipose depots contain remnant brown adipocytes interspersed among white adipocytes, and disturbances of brown with respect to white adipocyte biology have been implicated in highly active antiretroviral therapy (HAART)-induced lipo matosis. Brown adipocytes express the uncoupling protein-1 (UCP1) and contain a large number of mitochondria, potential targets of HAART toxicity. The aim of this study was to evaluate the effects of reverse transcriptase inhibitors (RTIs) on primary brown adipocytes differentiated in culture.

Design and methods

We analysed the effects of RTIs, nucleoside analogues (NRTIs: stavudine, zidovudine, didanosine and lamivudine) and non-nucleoside analogues (NNRTIs: nevirapine and efavirenz), on differentiation, mitochondrial biogenesis and gene expression in brown adipocytes.

Results

None of the NRTIs altered brown adipocyte differentiation whereas NNTRIs had differing effects. Efavirenz blocked lipid deposition and expression of adipose marker genes but nevirapine induced lipid accumulation and adipose gene expression, promoted mitochondrial biogenesis and increased UCP1. Stavudine, zidovudine and didanosine reduced mitochondrial DNA (mtDNA) content. However, mitochondrial genome expression was only impaired in didanosine-treated adipocytes. Stavudine, but not zidovudine, induced expression of the mitochondrial transcription factors and this may explain compensatory mechanisms for the depletion of mtDNA by up-regulating mtDNA transcription. Stavudine caused a specific induction of UCP1 gene expression through direct interaction with a retinoic acid-dependent pathway.

Conclusions

Specific disturbances in brown adipocytes in adipose depots may contribute to HAART-induced lipomatosis. Mitochondrial depletion does not appear to be the only mechanism explaining adverse effects in brown adipocytes because there is evidence of compensatory mechanisms that maintain mtDNA expression, and the expression of the UCP1 gene is specifically altered.
Mitochondrial Dna Depletion in HIV-Infected Patients is More Pronounced with Chronic Hepatitis C and Enhanced following Treatment with Pegylated Interferon plus Ribavirin
Authors: de Mendoza Carmen , Sánchez-Conde Matilde , Timmermans Eveline , Buitelaar Marije , de Baar Michel P , Gonzalez-Lahoz Juan , Soriano Vincent
Published: April 2022

Background

Mitochondrial DNA (mtDNA) damage seems to be responsible for many of the toxicities associated with the long-term use of nucleoside analogues in HIV-infected patients. These adverse effects, mainly lipoatrophy, seem to be even more pronounced in subjects with hepatitis C virus (HCV) co-infection. However, there is no information about a possible additive effect of HCV on mtDNA depletion nor about the impact of ribavirin use in HIV/HCV-coinfected individuals.

Patients and methods

mtDNA was measured in peripheral blood mononuclear cells (PBMC) collected from 192 individuals classified into 4 groups: HIV-neg/HCV-neg (control group, n=11), HIV-pos/HCV-neg (56), HIV-neg/HCV-pos (18) and HIV-pos/HCV-pos (107). A duplex real-time NASBA assay was used to quantify mtDNA on maximal platelet-depleted specimens and all experiments were run in duplicate. The mtDNA copy number per cell was estimated taking as reference the nuclear DNA copy number.

Results

The mean mtDNA values in the control group was 757 copies/cell, while it was 428, 349 and 296 for HIV-pos, HCV-pos and HIV/HCV-coinfected individuals, respectively ( P<0.001 for all groups relative to the control group). No significant differences were observed when comparing patients with HIV or HCV infections alone, but coinfected individuals showed a lower mtDNA copy number than patients infected with HIV ( P<0.001) or with HCV ( P=0.089). In a subset of 18 patients with HIV/HCV-coinfection, treatment with pegylated interferon plus ribavirin produced a further reduction in mtDNA (mean value, 189 copies/cell; P=0.009).

Conclusions

HIV and HCV may independently cause mtDNA depletion in PBMC. Coinfection may result in more pronounced mtDNA depletion. The administration of interferon plus ribavirin may further enhance mtDNA depletion. These findings may explain the greater risk of lipoatrophy of antiretroviral therapy in HIV-infected patients with HCV coinfection and why anti-HCV therapy may aggravate this effect.
Facial Fat Volume in HIV-Infected Patients with Lipoatrophy
Authors: Yang Yong , Sitoh Yih-Yian , Tha Naing Oo , Paton Nicholas I
Published: April 2022

Background

The volume and anatomical distribution of facial fat depletion in HIV related lipoatrophy have not been properly quantified. We aimed to determine the extent and distribution of facial fat loss in HIV-infected patients with lipoatrophy and whether this differs from wasting.

Design

We studied HIV-infected patients with clinically defined moderate to severe lipoatrophy without wasting ( n=15), clinically defined wasting (>10% weight loss and recent opportunistic infection) with no previous reported lipodystrophy ( n=15), and controls without lipodystrophy or wasting ( n=15). Facial MRI scans were performed, and areas of fat were manually traced bilaterally in all individual image slices and summed to calculate volume in anatomically defined regions of interest.

Results

Patients with lipoatrophy had lower fat volumes in the temporal region (8.9 ±9.4 vs 20.5 ±7.2ml; P<0.001), cheek region (25.6 ±24.9 vs 55.5 ±15.0 ml; P<0.001), periorbital region (1.9 ±1.0 vs 2.7 ±1.0 ml; P=0.09) and buccal fat pad (13.4 ±9.4 vs 21.8 ±9.8 ml; P=0.030) compared with controls. Patients with wasting had temporal, cheek, periorbital and buccal fat pad volumes (10.4 ±6.7 ml, 34.0 ±14.8 ml, 1.4 ±1.1 ml and 13.1 ±4.6 ml respectively) that were lower than controls (all P<0.01) but similar to lipoatrophy patients (all P>0.5).

Conclusions

Facial fat depletion in lipoatrophy is substantial (approximately 50% volume loss) and involves superficial and deep fat (buccal fat pad). The distribution and volume of fat change is similar to that seen in wasting. Given the extent of the changes, complete surgical correction is unlikely to be possible and hence emphasis should be placed on prevention of
The HIV Protease Inhibitors Saquinavir, Ritonavir, and Nelfinavir Induce Apoptosis and Decrease Barrier Function in Human Intestinal Epithelial Cells
Authors: Bode Hagen , Lenzner Luzie , Kraemer Oliver Holger , Kroesen Anton Josef , Bendfeldt Kerstin , Schulzke Jörg Dieter , Fromm Michael , Stoltenburg-Didinger Gisela , Zeitz Martin , Ullrich Reiner
Published: April 2022

Background and aims

Diarrhoea is a frequent adverse effect of HIV protease inhibitors (PIs) which may be due to intestinal barrier disruption. We investigated whether tight junction dysregulation, apoptosis or necrosis are responsible for this epithelial damage.

Methods

Saquinavir, nelfinavir, and ritonavir were added to the mucosal or serosal side of HT-29/B6 colon cell monolayers. Transepithelial resistance was monitored for 72 h to assess epithelial barrier function. Apoptosis and necrosis were investigated by light and electron microscopy and quantified by nucleosome ELISA and LDH measurement, respectively. Tight junction components were analysed by Western blots of occludin and zonula occludens. Apoptosis induction in normal human intestinal epithelium was examined by measurement of poly(ADP-ribose) polymerase (PARP) cleavage in Western blots of mucosal tissue explants cultured with PIs for 24 h.

Results

HIV PIs decreased transepithelial resistance by more than 44% in HT-29/B6 monolayers. Histology revealed massive apoptotic body formation but no evidence for necrosis after PI treatment. Correspondingly, LDH release was lower than 0.2%/h of total LDH, independent of PI treatment, and nucleosomes were increased up to 22-fold after drug treatment versus control. Occludin and zonula occludens-1 expression in the membrane were not diminished. PARP cleavage increased in normal human intestinal tissue treated with PIs.

Conclusions

PI-induced barrier disruption in intestinal epithelial cells is not due to necrosis or tight junction alterations, but to induction of massive apoptosis which may lead to leak-flux diarrhoea in vivo. Our findings suggest that induction of apoptosis by PIs could have potential for antitumour therapy.
Treating Hepatitis C in HIV-Infected Patients: Where are We Now?
Authors: Perronne Christian
Published: April 2022

The 1st International Workshop on HIV and Hepatitis Co-infection was held in Noordwijk, the Netherlands, 2–4 December 2004. This report outlines issues raised at the workshop concerning the treatment of hepatitis C in HIV-coinfected patients. Clinical trials using pegylated interferon in combination with ribavirin have provided positive outcomes, although clinicians still face a number of challenges in learning how best to use these drugs.
Long-Term Lamivudine Monotherapy in Renal-Transplant Recipients with Hepatitis-B-Related Cirrhosis
Authors: Viganò Mauro , Colombo Massimo , Aroldi Adriana , Lunghi Giovanna , Manenti Elena , Ponticelli Claudio , Lampertico Pietro
Published: April 2022

Background

Chronic hepatitis B virus (HBV) infection is an important cause of morbidity and mortality in renal-transplant recipients. The aim of the study was to assess the efficacy and safety of long-term lamivudine monotherapy in renal-transplant recipients with HBV-related cirrhosis.

Methods

Seventeen such patients [median age: 45 years; 7 with hepatitis B e antigen (HBeAg)] received daily oral doses of 75–150 mg lamivudine for a median of 48 (range 11–81) months. All patients had baseline serum levels of HBV DNA of over 6 log copies per ml and of 10 alanine transaminase (ALT) of over 1.5 times the upper normal limit (UNL). Clinical lamivudine resistance was defined as a rebound of serum HBV DNA above 5.3 log 10 copies per ml and of serum ALT of over 1.5 times the UNL in patients who initially responded with HBV DNA levels of less than 5.3 log copies per ml and normal ALT 10 values. Controls were 14 renal-transplant patients (median age 44 years; 3 with HBeAg) with HBV-related cirrhosis, naive to any anti-HBV therapy, followed for 58 months (4–135).

Results

Thirteen (77%) treated patients had a persistent response throughout the study period, including three (18%) who developed genotypic resistance, compared with none of the untreated controls (77% versus 0%, P<0.0001). Four (23%) developed clinical resistance. Two of three patients with initially decompensated cirrhosis had a durable response and clinical improvement compared with the transient responder, whose liver function worsened following lamivudine resistance. Two responders developed chronic rejection requiring chronic haemodialysis. Overall, one treated patient developed liver-related complications, compared with eight untreated controls (6% versus 57%, P<0.01).

Conclusions

Most renal-transplant patients treated with lamivudine achieved a rapid and durable suppression of HBV, which substantially lowered the risk of liver decompensation and death.
In vitro Phenotypic Susceptibility to Nucleoside Reverse Transcriptase Inhibitors of HIV-2 Isolates with the Q151M Mutation in the Reverse Transcriptase Gene
Authors: Damond Florence , Collin Gilles , Matheron Sophie , Peytavin Gilles , Campa Pauline , Delarue Séverine , Taieb Audrey , Bénard Antoine , Chêne Geneviève , Brun-Vézinet Françoise , Descamps Diane ,   for the French ANRS HIV-2 Cohort (ANRS CO 5 VIH-2)
Published: April 2022

In HIV-2 infection, many studies have reported a high frequency of selection of the Q151M mutation, but its impact on phenotypic susceptibility of HIV-2 isolates remains unclear. Four HIV-2 infected patients from the French ANRS HIV-2 cohort, with evidence of Q151M mutation in both plasma and available peripheral blood mononuclear cells (PBMCs) co-cultivated supernatants, were selected. In vitro phenotypic susceptibilities to different nucleoside reverse transcriptase inhibitors (NRTIs) were determined using a PBMC assay. In HIV-2 isolates, the Q151M mutation alone impacts only the phenotypic susceptibility to stavudine and abacavir. A decrease in susceptibility to all NRTIs was observed when Q151M was selected with V111I, a mutation of unknown impact on HIV-1 resistance. Clinical relevance of these phenotypic susceptibility results needs to be evaluated in HIV-2 treated patients.
Simplification Therapy with Once-Daily Didanosine, Tenofovir and Efavirenz in HIV-1-Infected Adults with Viral Suppression Receiving a More Complex Antiretroviral Regimen: Final Results of the EFADITE Trial
Authors: Barrios Ana , Negredo Eugenia , Domingo Pere , Estrada Vicente , Labarga Pablo , Asensi Víctor , Morales Dolores , Santos Jesús , Clotet Bonaventura , Soriano Vincent ,   for the EFADITE Study Group
Published: April 2022

Background

High pill burden and side effects often impact on the long-term success of highly active anti-retroviral therapy (HAART), which has led clinicians to search for more convenient regimens.

Patients and methods

A prospective, multicentre, open, comparative study in which HIV-1-infected patients on HAART and with plasma HIV-1 RNA <50 copies/ml for longer than 6 months were switched to tenofovir, didanosine and efavirenz (QD arm) or remained on the same treatment regimen (control arm). Patients with grade 4 toxicities or plasma HIV-1 RNA values repeatedly >1000 copies/ml discontinued the study.

Results

A total of 390 patients were included in the trial (309 in the QD arm and 81 in the control arm). The main baseline characteristics were well balanced between groups. In the QD arm, 41% of patients received high (standard) didanosine doses and 59% received reduced doses. At 12 months, plasma HIV-1 RNA <400 copies/ml was attained in 66% of QD patients and 73% of controls in the intent-to-treat (ITT) analysis ( P=NS). However, the number of individuals with HIV-1 RNA <400 copies/ml in the QD arm was 56% versus 71% when comparing the use of high versus low didanosine doses ( P=0.007). Treatment discontinuation occurred in 87 QD cases (28%) and 17 controls (21%). Twenty QD individuals (6.5%) and 2 controls (2.5%) discontinued because of virological failure ( P=NS). The median CD4+ cell count change at 12 months was -26 and +27 cells/μl in QD patients and controls, respectively ( P=0.001). In individuals who attained HIV-1 RNA <400 copies/ml, CD4+ cell changes were -25 and +15 cells/μl in QD patients and controls, respectively ( P=0.001). Moreover, CD4+ cell declines in the QD arm were significantly greater in patients taking high versus low didanosine doses (-59 versus -15 cells/μl; P=0.04). The lipid profile improved significantly in the QD arm, particularly in patients who were on protease inhibitors prior to simplification.

Conclusions

Simplification to didanosine–tenofovir–efavirenz provides a virological suppression rate at 12 months similar to that seen in patients who do not change therapy, as long as low didanosine doses are administered. Decreases in CD4+ cell levels in patients in the QD arm (especially decreases seen with high didanosine doses) and dyslipidaemias along with less convenient pill burden and schedules in controls were the main long-term concerns for each option.
Arterial Stiffness in HIV-Infected Patients Receiving Highly Active Antiretroviral Therapy
Authors: Sevastianova Ksenia , Sutinen Jussi , Westerbacka Jukka , Ristola Matti , Yki-Järvinen Hannele
Published: April 2022

HIV-infected patients receiving highly active antiretroviral therapy (HAART) are at increased risk of cardiovascular events. Reported non-invasive techniques for assessment of blood pressure in this population have been limited to sphygmomanometry. The present cross-sectional study investigated the impact of antiretroviral therapy and the HAART-associated lipodystrophy on aortic blood pressure conditions and arterial stiffness in HAART-treated lipodystrophic ( n=42) and non-lipodys-trophic ( n=17) patients. Pulse wave analysis, novel to this population, was used to evaluate measures of arterial stiffness, including the heart rate corrected augmentation index, AgIHR. Results indicated no significant difference between the study groups in peripheral or aortic blood pressure and AgIHR. Significant correlates of AgIHR included age ( P=0.003), duration of antiretroviral therapy ( P=0.020), lamivudine therapy ( P=0.015) and ritonavir therapy ( P=0.016) as well as cumulative exposure to protease inhibitors ( P=0.030). Time since HIV diagnosis, severity of immunodeficiency or presence of HAART-associated lipodystrophy bore no relationship to AgIHR. In multivariate analysis, duration of antiretroviral therapy ( P=0.046), cumulative exposure to nucleoside reverse transcriptase inhibitors ( P=0.032) and to protease inhibitors ( P=0.011) were identified as independent factors predicting AgIHR. Prolonged antiretroviral treatment, thus, delineates as a risk factor for systemic arterial stiffness and the associated cardiovascular mortality.
HIV Protease Inhibitors Prevent Mitochondrial Hyperpolarization and Redox Imbalance and Decrease Endogenous Uncoupler Protein-2 Expression in Gp120-Activated Human T Lymphocytes
Authors: Matarrese Paola , Tinari Antonella , Gambardella Lucrezia , Mormone Elisabetta , Narilli Piero , Pierdominici Marina , Cauda Roberto , Malorni Walter
Published: April 2022

It has been demonstrated that HIV protease inhibitors (PIs) are able to inhibit apoptosis of both infected and uninfected T cells. It was hypothesized that the mechanisms underlying this effect are associated with a specific activity of these drugs against mitochondrial modifications occurring in the execution phase of apoptosis. In this work, we investigated the activity of PIs towards the early changes occurring in mitochondrial membrane potential in freshly isolated uninfected human T lymphocytes sensitized to CD95/Fas-induced physiological apoptosis via pre-exposure to HIV envelope protein gp120. The results obtained clearly indicate that PIs are capable of hindering early morphogenetic changes bolstering T cell apoptosis, that is, cell polarization and mitochondrial hyperpolarization. The target effect on mitochondria appeared to be characterized by a specific activity of PIs in the maintenance of their homeostasis either in intact cells or in cell-free systems, that is, isolated mitochondria. PIs seem to act as boosters of mitochondrial defense mechanisms, including modulation of endogenous uncouplers. These results add new insights in the field of PI mitochondrial toxicity mechanisms and pharmacological perspectives for the use of these drugs in the control of immune system homeostasis.
Uridine in the Prevention and Treatment of Nrti-Related Mitochondrial Toxicity
Authors: Walker Ulrich A , Venhoff Nils
Published: April 2022

Long-term side effects of antiretroviral therapy are attributed to the mitochondrial (mt) toxicity of nucleoside analogue reverse transcriptase inhibitors (NRTIs) and their ability to deplete mtDNA. Studies in hepatocytes suggest that uridine is able to prevent and treat mtDNA depletion by pyrimidine NRTIs [zalcitabine (ddC) and stavudine (d4T)] and to fully abrogate hepatocyte death, elevated lactate production and intracellular steatosis. Uridine was also found to improve the liver and haematopoietic toxicities of zidovudine (AZT), which are unrelated to mtDNA depletion, and to prevent neuronal cell death induced by ddC. Most recently, uridine was found to prevent the onset of a lipoatrophic phenotype (reduced intracellular lipids, increased apoptosis, mtDNA depletion and mt depolarization) in adipocytes incubated long-term with d4T and AZT. Various steps of mt nucleoside utilization may be involved in the protective effect, but competition of uridine metabolites with NRTIs at polymerase y or other enzymes is a plausible explanation. Pharmacokinetic studies suggest that uridine serum levels can be safely increased in humans to achieve concentrations which are protective in vitro (50–200 μM). Uridine was not found to interfere with the antiretroviral activity of NRTIs. Mitocnol, a sugar cane extract which effectively increases uridine in human serum, was beneficial in individual HIV patients with mt toxicity and is now being tested in placebo-controlled randomized trials. Until these data become available, the risk-benefit calculation of using uridine should be individualized. The current safety data justify the closely monitored use of uridine in individuals who suffer from mt toxicity but who cannot be switched to less toxic NRTIs.
Increased Antiretroviral Potency by the Addition of Enfuvirtide to a Four-Drug Regimen in Antiretroviral-Naive, HIV-Infected Patients
Authors: Moltó José , Ruiz Lidia , Valle Marta , Martinez-Picado Javier , Bonjoch Ana , Bravo Isabel , Negredo Eugenia , Heilek-Sneider Gabrielle M , Clotet Bonaventura
Published: April 2022

Objective

To assess if enfuvirtide (ENF) increases antiviral activity of a highly active four-drug antiretroviral (ARV) regimen containing lopinavir/ritonavir, efavirenz, lamivudine and tenofovir in ARV-naive, HIV-infected patients.

Methods

Pilot study in ARV-naive, HIV-infected patients with viral load (VL) >10,000 copies/ml and no documented resistance to any of the study drugs. Patients were randomized to receive ENF (ENF Group) or not (Control Group) in combination with lopinavir/ritonavir, efavirenz, lamivudine and tenofovir as a backbone. The primary endpoint was to assess differences in the HIV-1 RNA decay rate during the first phase of viral decay. VL and treatment adherence were measured at baseline, every 6 h during the first 3 days, and once daily from day 3 to 6. Individual HIV-1 RNA decay rates were obtained using a non-linear least squares regression model.

Results

Eight subjects were included in each study group. Mean (sd) baseline VL was 4.98 (0.38) log10 copies/ml in the ENF Group and 5.10 (0.49) log10 copies/ml in the Control Group ( P=0.607). Baseline CD4+ cell count was 463 (306) and 362 (225) cells/mm3 in the ENF and the Control Group, respectively ( P=0.484). First phase HIV-1 RNA decay rate was 0.802 (0.127) d-1 in the ENF Group and 0.624 (0.182) d-1 in the Control Group ( P=0.045). By day 6, VL was 3.55 (0.40) and 3.92 (0.36) log10 copies/ml in the ENF and the Control Group, respectively ( P=0.079).

Conclusion

The addition of ENF increased the antiviral potency of a highly active four-drug ARV regimen by 28.5% in ARV-naive, HIV-infected patients. The clinical impact of this finding should be assessed.