Studio longitudinale dell'evoluzione del tropismo per X4/R5, dei livelli di HIV-1 DNA cellulare, dei parametri immunologici e della viremia residua in una popolazione di pazienti naive trattati con successo

Background: Nowadays HIV-1 RNA levels and CD4+ T lymphocyte counts are the standard markers used in clinical practice for the management of HIV infection. However the evolution of HIV infection can be monitored also by measuring HIV-DNA and this measurement can be determined in PBMCs, even during powerful and prolonged antiretroviral therapy. In successfull treated HIV-1 patients, viral load is undetectable and the strategies for managing long-term side effects may involve a new class of antiretroviral-like CCR5 antagonists. Moreover the dynamics and the influence of viral tropism on the course of HIV-1 infection in subjects exposed to antiretroviral therapy are not fully understood. Then the evolution and determination of HIV-1 tropism based on cellular DNA sequence could be useful for patients with a successfully suppressed plasma viral load. Aims: In this study we aimed to determine whether the HIV-1 tropism for CXCR4 or CCR5 correlates with residual viraemia, cellular HIV-1 DNA load and CD4+ count; moreover, we evaluated if exist a correlation between baseline and follow-up HIV-1 DNA levels with residual viraemia, baseline plasma HIV-1 RNA, and the condition of primary or chronic HIV infection at the start of antiretroviral therapy. Methods: In the CAVeAT, that is a prospective cohort of HIV-infected patients enrolled starting from 2004 in five infectious diseases units in Northeastern Italy (Veneto region), we retrospectively selected two subgroup of patients (cohort I and cohort II); they were a subset of subjects achieving virological suppression within 6 months after initiation of first-line therapy and maintaining plasma HIV RNA levels < 50 copies/ml, without virological failures, until evaluation at the follow-up time points. In order to be included in the our study, the patients needed to be naïve and treated with effective antiretroviral therapy. None of the patients were treated with CCR5 antagonists. The cohort I consisted on 219 patients with median follow-up time of 3 years (T0, T1, T2) while the cohort II was represented of 181 patients with median follow-up times of 4 years (T0, T1, T2, T3). Genotypic analysis of viral tropism was performed on PBMCs throught the sequencing of V3 loop of gp120; the generated sequences were interpreted using the bioinformatic tool Geno2pheno coreceptor while proviral DNA was quantified by Real-Time PCR using TaqMan probes. Results: In the cohort I, HIV-1 DNA, CD4+ count and plasma viraemia were available from all 219 patients at T0 and T1, and in 86 subjects at T2, while tropism determinations were available from 109 subjects at T0, 219 at T1, and from 86 subjects at T2. The results showed that achieving a residual viraemia < 2.5 cp/ml at T1 correlated with having the same condition at T2 and that there was a positive correlation between To and T1 -T2 tropism. X4 tropism at T1 negatively correlated with the possibility of achieving viraemia < 2.5 cp/ml at T2 while a positive correlation between viremic suppression and R5 coreceptor affinity was found. In 181 patients of the cohort II, viroimmunological data were collected at baseline (T0) and at two follow-up time points (T1, T2); in a subgroup of 70 subjects, we evaluated also a third follow-up time point (T3). We observed that high baseline plasma HIV-1 RNA values positive correlated with high levels of HIV-1 DNA at T0, T1, T2, T3 and negative correlated with residual viraemia at T1, T2, T3; having high levels of HIV-1 DNA at T0 positive correlated with high values at T1, T2, T3 and negatively correlated with achieving residual viraemia. Primary infection was associated with lower probability of having high HIV-1 DNA levels at T1, T2, T3 and with a higher probability of achieving residual viraemia at T1 and T3, with respect to chronic infection. Conclusions: The tropism of archived virus was stable during an effective treatment, although a low percentage of patients switched over time. R5 tropism and its stability were related to achieving and maintaining viraemia < 2.5 copies/ml, in treatment responder patients, suggesting a relation among viral tropism and response to treatment in the long term. Moreover, we demonstrated a strong and long-lasting correlation between viral load and cellular HIV-1 DNA before and after the start of HAART and that cellular HIV-1 DNA is closely related to residual viraemia over long-term follow-up of ART responders, particularly when treated during primary infection

Prevalence, molecular epidemiology and intra-hospital acquisition of Klebsiella pneumoniae strains producing carbapenemases in an Italian teaching hospital from January 2015 to September 2016.

Objectives: We described Klebsiella pneumoniae producing carbapenemase (CPKP) spread from 01/01/2015 to 13/09/16 in a tertiary level hospital. Methods: The first positive surveillance rectal swab (SRS) or clinical sample (CS) collected in the medical department (MD), surgical department (SD) and intensive care department (ICD) were included in the study. A validated in-house Real-Time PCR method was used to detect carbapenemases; multilocus sequence typing (MLST) was used for further characterization of the strains. Results: 21535 patients were included: 213 CPKP strains from surveillance rectal swab (SRS) and 98 from clinical samples (CS) were collected. The percentage of CPKP detected in SRS with respect to CS increased in the medical MD from 2015 to 2016 (p = 0.01) and in ICD from 2012 to 2015 (p = 0.0001), while it decreased in SD from 2014 to 2016 (p = 0.003); 68.5% of the positive SRS had a previous negative SRS; CPKP was more frequently identified in CS than in SRS in MD. Twelve strains harboured more than one carbapenemase gene. Many other species harbouring a carbapenemase gene were collected. Conclusions: MDs need more inclusive surveillance criteria. The late detection of positive SRS underlined the risk of colonization during hospitalization

Viral infections of the central nervous system in elderly patients: a retrospective study

Summary Objectives Very few data exist on viral meningitis and encephalitis in elderly patients (>65 years old). Methods This study investigated the detection of herpes simplex virus (HSV), varicella zoster virus (VZV), human herpes virus 6 (HHV-6), HHV-7, HHV-8, cytomegalovirus (CMV), Epstein–Barr virus (EBV), enterovirus (EV), human adenovirus (HAdV), human parechoviruses (HPeVs), and tick-borne encephalitis virus (TBEV) through real-time PCR (RT-PCR) in patients >65 years old who had cerebrospinal fluid (CSF) tested for a suspected central nervous system infection. Results A total of 2868 RT-PCRs were performed on 502 CSF samples. Overall, 65 positive RT-PCRs were found: 23 for HSV (35.4% of positives), 15 for EV (23.1% of positives), 14 for EBV (21.5% of positives), 12 for VZV (18.5% of positives), and one for CMV (1.5% of positives). A positive RT-PCR in CSF was detected in 24 (17.4%) patients aged ≥80 years and in 35 (9.6%) patients aged 65–79 years ( p =0.02). VZV was more frequently detected in the oldest subjects (5.9% vs. 1.6%, p =0.03). Conclusions HSV was the most common viral aetiology identified in the study, with VZV infection being recognized more frequently in those patients aged ≥80 years

A stable CC-chemokine receptor (CCR)-5 tropic virus is correlated with the persistence of HIV RNA at less than 2.5 copies in successfully treated naïve subjects

BACKGROUND: To determine if tropism for CXCR4 or CCR5 correlates with cellular HIV DNA load, residual viraemia and CD4 count in 219 successfully treated naive subjects with HIV infection enrolled in five infectious diseases units in Northeastern Italy. METHODS: A subset of subjects, achieving plasma HIV RNA level <50 copies/ml after initiation of first-line therapy and maintaining it until follow-up time points, was retrospectively selected from a prospective cohort. Blood samples were collected before the beginning of therapy (T0), at the first follow-up time (T1) and, when available, at a second (T2) follow-up time. RESULTS: HIV DNA, CD4 count and plasma viraemia were available from all 219 patients at T0 and T1, and in 86 subjects at T2, while tropism determinations were available from 109 subjects at T0, 219 at T1, and from 86 subjects at T2. Achieving residual viraemia <2.5 copies/ml at T1 correlated with having the same condition at T2 (p = 0.0007). X4 tropism at T1 was negatively correlated with the possibility of achieving viraemia<2.5 copies/ml at T2 (p = 0.0076). T1-T2 tropism stability was significant (p <0.0001). T0 tropism correlated with T1 and T2 tropism (p < 0.001); therefore the stability of the tropism over the two follow-up periods was significant (p = 0.0003). An effective viremic suppression (viraemia<2.5 copies/ml) correlated with R5 coreceptor affinity (p= 0.047). CONCLUSIONS: The tropism of archived virus was stable during an effective treatment, with 15-18% of subjects switching over time, despite a viraemia<50 copies/ml. R5 tropism and its stability were related to achieving and maintaining viraemia<2.5 copies/ml

Daclatasvir plasma level and resistance selection in HIV patients with hepatitis C virus cirrhosis treated with daclatasvir, sofosbuvir, and ribavirin

ObjectivesEffective treatment with direct-acting antiviral drugs against hepatitis C virus (HCV) is a medical need in cirrhotic HIV–HCV co-infected patients.MethodsThis study investigated the plasma levels of daclatasvir (DCV) and ribavirin (RBV) in HIV–HCV co-infected subjects treated with DCV, sofosbuvir, and RBV. Drug concentrations were quantified using validated high-performance liquid chromatography methods with ultraviolet detection. The HCV non-structural protein 5A and non-structural protein 5B coding regions were analyzed by population-based sequencing.ResultsDCV was dosed at week 4 and at week 8 of treatment, and RBV at week 8. One patient had the lowest DCV level, corresponding to 32.7% of the overall median value of the other patients at week 4 and about 40% at week 8. The Y93H variant was detected in this subject at weeks 8, 16, and 20 of treatment, but not before treatment or at day 2, and the patient experienced virological failure. Another subject with the Y93H variant at baseline and appropriate DCV levels had HCV RNA <12 IU/ml at week 12 and undetectable at week 16.ConclusionsSub-optimal DCV drug levels allow the selection of resistance-associated variants and fail to contribute to antiviral activity. No definite reason for the low DCV level was found. Quantifying the drug is suggested in difficult-to-treat patients

Liver stiffness is not associated with short- and long-term plasma HIV RNA replication in immunocompetent patients with HIV infection and with HIV/HCV coinfection

Background:Human immunodeficiency virus (HIV) may be directly responsible for liver damage but there are contrasting data regarding the influence of detectable plasma viremia. We analyzed the influence of plasma HIV RNA (pHIV) detectability and of other clinical and viro-immunological variables on liver stiffness (LS) measurement in adult immunocompetent HIV-monoinfected patients and in patients coinfected with hepatitis C virus (HCV). Methods: Logistic regression analysis was performed using the value of LS>7.1 kPa as the dependent variable. A linear regression model was applied using LS measurement after log 10 transformation (lkpa) as the dependent variable and we analyzed the predicted values versus the observed lkpa values; pHIV was classified as detectable or undetectable in the 12- and 36-month study periods before LS measurement. Results: We studied 251 patients (178 with HIV monoinfection), most of whom were on antiviral treatment; 36-month study time was available for 154 subjects. The mean CD4+ cell count was 634 cells/mm3 in HIV-monoinfected patients and 606 cells/mm3 in coinfected patients. No difference in LS was found between patients with detectable or undetectable pHIV in either the 12- or the 36-month study period before transient elastography. The mean LS was higher in HIV/HCV coinfected patients (P<0.0001) than in the HIV-monoinfected subjects; lkpa was positively correlated with HCV coinfection (P<0.0001) and aspartate aminotransferase levels (P<0.0001). Detectable pHIV failed to reach significance. Eight HIV-monoinfected patients had a predicted LS measurement lower than the observed one, while eight patients had the opposite result. Conclusion: LS was not correlated with ongoing HIV replication during the 12- and 36-month study periods in immunocompetent HIV-monoinfected and HIV/HCV-coinfected patients

Role of pretreatment variables on plasma HIV RNA value at the sixth month of antiretroviral therapy including all first line drugs in HIV na\uefve patients: A path analysis approach

We investigated the conditioning roles of viral tropism and other variables on plasma HIV RNA levels after 6 months of combination antiretroviral therapy (cART) in an HIV-infected Italian naïve population using regression tree, random forest regression, and path analysis (PA). Patients in this multicenter observational study were treated with all antiviral drugs that are currently recommended as first-line therapies

Anal and oral human papillomavirus (HPV) infection in HIV-infected subjects in northern Italy: a longitudinal cohort study among men who have sex with men

<p>Abstract</p> <p>Background</p> <p>A study including 166 subjects was performed to investigate the frequency and persistence over a 6-month interval of concurrent oral and anal Human Papillomavirus (HPV) infections in Human Immunodeficiency Virus (HIV)-infected men who have sex with men (MSM).</p> <p>Methods</p> <p>Patients with no previously documented HPV-related anogenital lesion/disease were recruited to participate in a longitudinal study. Polymerase chain reaction (PCR) was performed to detect HPV from oral and anal swabs and to detect Human Herpes Virus 8 (HHV-8) DNA in saliva on 2 separate specimen series, one collected at baseline and the other collected 6 months later. A multivariate logistic analysis was performed using anal HPV infection as the dependent variable versus a set of covariates: age, HIV plasma viral load, CD4+ count, hepatitis B virus (HBV) serology, hepatitis C virus (HCV) serology, syphilis serology and HHV-8 viral shedding. A stepwise elimination of covariates with a p-value > 0.1 was performed.</p> <p>Results</p> <p>The overall prevalence of HPV did not vary significantly between the baseline and the follow-up, either in the oral (20.1 and 21.3%, respectively) or the anal specimens (88.6 and 86.3%). The prevalence of high-risk (HR) genotypes among the HPV-positive specimens was similar in the oral and anal infections (mean values 24.3% and 20.9%). Among 68 patients with either a HR, low-risk (LR) or undetermined genotype at baseline, 75% had persistent HPV and the persistence rates were 71.4% in HR infections and 76.7% in LR infections. There was a lack of genotype concordance between oral and anal HPV samples. The prevalence of HR HPV in anus appeared to be higher in the younger patients, peaking (> 25%) in the 43-50 years age group. A decrease of the high level of anal prevalence of all genotypes of HPV in the patients > 50 years was evident. HHV-8 oral shedding was positively related to HPV anal infection (p = 0.0046). A significant correlation was found between the persistence of HHV-8 shedding and HIV viral load by logistic bivariate analysis (Odds Ratio of HHV-8 persistence for 1-log increase of HIV viral load = 1.725 ± 0.397, p = 0.018).</p> <p>Conclusions</p> <p>A high prevalence of HPV infection was found in our cohort of HIV-infected MSM, with a negative correlation between anal HPV infection and CD4 cell count.</p

Studio longitudinale dell'evoluzione del tropismo per X4/R5, dei livelli di HIV-1 DNA cellulare, dei parametri immunologici e della viremia residua in una popolazione di pazienti naive trattati con successo

Background: Nowadays HIV-1 RNA levels and CD4+ T lymphocyte counts are the standard markers used in clinical practice for the management of HIV infection. However the evolution of HIV infection can be monitored also by measuring HIV-DNA and this measurement can be determined in PBMCs, even during powerful and prolonged antiretroviral therapy. In successfull treated HIV-1 patients, viral load is undetectable and the strategies for managing long-term side effects may involve a new class of antiretroviral-like CCR5 antagonists. Moreover the dynamics and the influence of viral tropism on the course of HIV-1 infection in subjects exposed to antiretroviral therapy are not fully understood. Then the evolution and determination of HIV-1 tropism based on cellular DNA sequence could be useful for patients with a successfully suppressed plasma viral load. Aims: In this study we aimed to determine whether the HIV-1 tropism for CXCR4 or CCR5 correlates with residual viraemia, cellular HIV-1 DNA load and CD4+ count; moreover, we evaluated if exist a correlation between baseline and follow-up HIV-1 DNA levels with residual viraemia, baseline plasma HIV-1 RNA, and the condition of primary or chronic HIV infection at the start of antiretroviral therapy. Methods: In the CAVeAT, that is a prospective cohort of HIV-infected patients enrolled starting from 2004 in five infectious diseases units in Northeastern Italy (Veneto region), we retrospectively selected two subgroup of patients (cohort I and cohort II); they were a subset of subjects achieving virological suppression within 6 months after initiation of first-line therapy and maintaining plasma HIV RNA levels < 50 copies/ml, without virological failures, until evaluation at the follow-up time points. In order to be included in the our study, the patients needed to be naïve and treated with effective antiretroviral therapy. None of the patients were treated with CCR5 antagonists. The cohort I consisted on 219 patients with median follow-up time of 3 years (T0, T1, T2) while the cohort II was represented of 181 patients with median follow-up times of 4 years (T0, T1, T2, T3). Genotypic analysis of viral tropism was performed on PBMCs throught the sequencing of V3 loop of gp120; the generated sequences were interpreted using the bioinformatic tool Geno2pheno coreceptor while proviral DNA was quantified by Real-Time PCR using TaqMan probes. Results: In the cohort I, HIV-1 DNA, CD4+ count and plasma viraemia were available from all 219 patients at T0 and T1, and in 86 subjects at T2, while tropism determinations were available from 109 subjects at T0, 219 at T1, and from 86 subjects at T2. The results showed that achieving a residual viraemia < 2.5 cp/ml at T1 correlated with having the same condition at T2 and that there was a positive correlation between To and T1 -T2 tropism. X4 tropism at T1 negatively correlated with the possibility of achieving viraemia < 2.5 cp/ml at T2 while a positive correlation between viremic suppression and R5 coreceptor affinity was found. In 181 patients of the cohort II, viroimmunological data were collected at baseline (T0) and at two follow-up time points (T1, T2); in a subgroup of 70 subjects, we evaluated also a third follow-up time point (T3). We observed that high baseline plasma HIV-1 RNA values positive correlated with high levels of HIV-1 DNA at T0, T1, T2, T3 and negative correlated with residual viraemia at T1, T2, T3; having high levels of HIV-1 DNA at T0 positive correlated with high values at T1, T2, T3 and negatively correlated with achieving residual viraemia. Primary infection was associated with lower probability of having high HIV-1 DNA levels at T1, T2, T3 and with a higher probability of achieving residual viraemia at T1 and T3, with respect to chronic infection. Conclusions: The tropism of archived virus was stable during an effective treatment, although a low percentage of patients switched over time. R5 tropism and its stability were related to achieving and maintaining viraemia < 2.5 copies/ml, in treatment responder patients, suggesting a relation among viral tropism and response to treatment in the long term. Moreover, we demonstrated a strong and long-lasting correlation between viral load and cellular HIV-1 DNA before and after the start of HAART and that cellular HIV-1 DNA is closely related to residual viraemia over long-term follow-up of ART responders, particularly when treated during primary infection.Background: Fino ad oggi nella pratica clinica la gestione dell'infezione da HIV-1 ha utilizzato come marcatori prognostici i livelli di RNA plasmatico e il numero dei linfociti CD4+. Tuttavia nei pazienti trattati, l'evoluzione dell'infezione da HIV può essere valutata anche misurando l'HIV-1 DNA nei reservoirs. In pazienti HIV-1 trattati con terapia di successo, la carica virale è non rilevabile e le strategie per la gestione degli effetti collaterali a lungo termine potrebbero considerare l'utilizzo di una nuova classe di antiretrovirali antagonisti del CCR5. Inoltre non è ancora stato del tutto chiarito come il tropismo virale e la sua evoluzione influenzino il decorso dell'infezione da HIV-1 in soggetti in terapia antiretrovirale. Quindi per valutare come gestire i pazienti trattati con successo nei quali la carica virale è soppressa, potrebbe essere utile determinare l'evoluzione del tropismo di HIV-1 sul DNA cellulare. Scopo: In questo studio di coorte costituita da pazienti naive trattati con successo abbiamo valutato se il tropismo per CXCR4 o CCR5 di HIV-1 correla con la viremia residua; inoltre abbiamo determinato se esiste una correlazione tra i livelli di HIV-1 DNA cellulare (al baseline e ai follow-up) con la viremia residua, l'HIV-1 RNA al baseline, e con il fatto di avere un'infezione acuta all'inizio della terapia antiretrovirale. Materiali e metodi: All'interno della coorte CAVeAT, che è una coorte prospettica di pazienti HIV positivi arruolati a partire dal 2004 da cinque Unità di Malattie Infettive della regione Veneto, sono stati retrospettivamente selezionati due sottogruppi di pazienti (coorte I e coorte II); sono tutti pazienti che hanno raggiunto una soppressione virologica entro i 6 mesi dopo il primo trattamento e hanno mantenuto livelli di HIV-1 RNA al di sotto di 50 copie/ml senza fallimento virologico per tutti i tempi di follow-up. Per essere eleggibili, i pazienti dovevano essere naive e poi essere trattati con una terapia antiretrovirale di successo. Nessuno dei pazienti doveva essere in trattamento con gli antagonisti del CCR5. La coorte I era costituita da 219 pazienti con una mediana di follow-up di 3 anni (T0, T1, T2) mentre la coorte II era rappresentata da 181 pazienti con una media di follow up di 4 anni (T0, T1, T2, T3). Le analisi genotipiche per il tropismo virale sono state eseguite su PBMCs mediante sequenziamento del loop V3 di gp120; le sequenze ottenute sono state interpretate utilizzando l'algoritmo bioinformatico Geno2pheno coreceptor mentre il DNA provirale è stato quantificato mediante Real-Time PCR utilizzando sonde TaqMan. Risultati: Nella coorte I, la valutazione dell'HIV-1 DNA, della conta dei CD4+ e della viremia è stata effettuata su tutti i 219 pazienti al T0 e T1, e su 86 soggetti al T2; il tropismo è stato determinato solo in 109 soggetti al T0, su tutti i 219 al T1, e in 86 pazienti al T2. I risultati hanno mostrato che il raggiungere una viremia residua al di sotto di 2.5 cp/ml al T1 correlava con il suo mantenimento al T2 e che c'era una correlazione positiva tra il tropismo al To e quello ai follow-up ( T1-T2 ). Avere un tropismo X4 al T1 correla negativamente con la possibilità di raggiungere una viremia residua al di sotto di 2.5 cp/ml al T2 mentre è stata trovata una correlazione positiva tra soppressione virologica e tropismo R5. Nei 181 pazienti della coorte II, i dati viro-immunologici sono stati eseguiti per tutti i soggetti al baseline (T0) e ai due tempi di follow-up (T1, T2); in un sottogruppo di 70 pazienti, abbiamo preso in considerazione anche un terzo tempo di follow-up (T3). Quindi abbiamo osservato che alti livelli di HIV-1 RNA al baseline correlavano positivamente con alti livelli di HIV-1 DNA al T0, T1, T2, T3 e negativamente con la viremia residua al T1, T2, T3; avere alti livelli di HIV-1 DNA al T0 correlava positivamente con alti livelli anche al T1, T2, T3 e negativamente con il raggiungimento della viremia residua al di sotto delle 2.5 copie/ml. Avere un'infezione acuta, piuttosto che cronica, all'inizio della terapia era associato con una più bassa probabilità di avere alti livelli di HIV-1 DNA al T1, T2, T3 e con una più alta probabilità di raggiungere una viremia residua al T1 e T3. Conclusioni: Il tropismo del virus archiviato è rimasto stabile durante tutto il periodo della terapia, sebbene in una bassa percentuale di pazienti sia cambiato nel tempo. In pazienti trattati con una terapia di successo, il tropismo R5 e la sua stabilità sono stati confrontati con il fatto di raggiungere e mantenere una viremia residua al di sotto di 2.5 copie/ml, suggerendo una relazione tra il tropismo virale e la risposta ad una terapia a lungo termine. Inoltre, è stata dimostrata una forte correlazione tra la carica virale e l'HIV-1 DNA prima e dopo l'inizio della terapia antiretrovirale (ART) e che l'HIV-1 DNA cellulare è strettamente connesso alla viremia residua nel follow-up a lungo termine nei soggetti rispondenti alla terapia, in modo particolare se trattati durante l'infezione acuta, ovvero il prima possibile