Absence of XMRV in Peripheral Blood Mononuclear Cells of ARV-Treatment Naïve HIV-1 Infected and HIV-1/HCV Coinfected Individuals and Blood Donors

<div><h3>Background</h3><p>Xenotropic murine leukemia virus-related virus (XMRV) has been found in the prostatic tissue of prostate cancer patients and in the blood of chronic fatigue syndrome patients. However, numerous studies have found little to no trace of XMRV in different human cohorts. Based on evidence suggesting common transmission routes between XMRV and HIV-1, HIV-1 infected individuals may represent a high-risk group for XMRV infection and spread.</p> <h3>Methodology/Principal Findings</h3><p>DNA was isolated from the peripheral blood mononuclear cells (PBMCs) of 179 HIV-1 infected treatment naïve patients, 86 of which were coinfected with HCV, and 54 healthy blood donors. DNA was screened for XMRV provirus with two sensitive, published PCR assays targeting XMRV <em>gag</em> and <em>env</em> and one sensitive, published nested PCR assay targeting <em>env</em>. Detection of XMRV was confirmed by DNA sequencing. One of the 179 HIV-1 infected patients tested positive for <em>gag</em> by non-nested PCR whereas the two other assays did not detect XMRV in any specimen. All healthy blood donors were negative for XMRV proviral sequences. Sera from 23 HIV-1 infected patients (15 HCV⁺) and 12 healthy donors were screened for the presence of XMRV-reactive antibodies by Western blot. Thirteen sera (57%) from HIV-1⁺ patients and 6 sera (50%) from healthy donors showed reactivity to XMRV-infected cell lysate.</p> <h3>Conclusions/Significance</h3><p>The virtual absence of XMRV in PBMCs suggests that XMRV is not associated with HIV-1 infected or HIV-1/HCV coinfected patients, or blood donors. Although we noted isolated incidents of serum reactivity to XMRV, we are unable to verify the antibodies as XMRV specific.</p> </div

Time to Switch to Second-line Antiretroviral Therapy in Children With Human Immunodeficiency Virus in Europe and Thailand.

Background: Data on durability of first-line antiretroviral therapy (ART) in children with human immunodeficiency virus (HIV) are limited. We assessed time to switch to second-line therapy in 16 European countries and Thailand. Methods: Children aged <18 years initiating combination ART (≥2 nucleoside reverse transcriptase inhibitors [NRTIs] plus nonnucleoside reverse transcriptase inhibitor [NNRTI] or boosted protease inhibitor [PI]) were included. Switch to second-line was defined as (i) change across drug class (PI to NNRTI or vice versa) or within PI class plus change of ≥1 NRTI; (ii) change from single to dual PI; or (iii) addition of a new drug class. Cumulative incidence of switch was calculated with death and loss to follow-up as competing risks. Results: Of 3668 children included, median age at ART initiation was 6.1 (interquartile range (IQR), 1.7-10.5) years. Initial regimens were 32% PI based, 34% nevirapine (NVP) based, and 33% efavirenz based. Median duration of follow-up was 5.4 (IQR, 2.9-8.3) years. Cumulative incidence of switch at 5 years was 21% (95% confidence interval, 20%-23%), with significant regional variations. Median time to switch was 30 (IQR, 16-58) months; two-thirds of switches were related to treatment failure. In multivariable analysis, older age, severe immunosuppression and higher viral load (VL) at ART start, and NVP-based initial regimens were associated with increased risk of switch. Conclusions: One in 5 children switched to a second-line regimen by 5 years of ART, with two-thirds failure related. Advanced HIV, older age, and NVP-based regimens were associated with increased risk of switch

Engagement in care among youth living with parenterally-acquired HIV infection in Romania

Transition from adolescent to adult care can be challenging for youth living with HIV. We describe the level of engagement in care and its impact on HIV outcomes in a group of patients infected in early childhood and followed-up through adolescence (15-19 years) and young adulthood (20-24 years) by the same medical team. We conducted a cohort study of youth born between 1985- 1993 and infected with HIV parenterally, followed at a single tertiary care centre. Individuals were followed from age 15 years or first clinic visit (whichever came last) until age 25 years or 30 Nov 2016 (whichever came first). A longitudinal continuum-of-care was constructed, categorizing individuals' status for each month between the ages of 15-25 years as: engaged in care (EIC); not in care (NIC: no clinic visits within past year); lost-to-follow-up (LTFU: NIC and did not return to clinic); or died. Those EIC were further subdivided by current CD4 count and viral load (VL). 545 individuals (52% male) were followed for 4775 person-years. 64 (12%) became LTFU and 27 (5%) died. At age 15, 92% were EIC, decreasing to 84% at age 20 and 74% at age 25. Of those EIC, HIV markers improved with age: 79% and 52% had a CD4≥200 cells/µl and VL<400 cps/ml at age 15; increasing to 86% and 73% at age 20 and 87% and 80% at age 25. 277 (55.5%) spent their entire adolescence EIC; this decreased to 202 (37%) for the years of young adulthood (p=0.0001). There were no observed demographic differences between those with continuous and intermittent engagement in care. We conclude that youth infected during early childhood tended to disengage from care, even when followed by the same medical team for a lengthy period of time. For those that did engage in care, HIV-related outcomes improved from adolescence through to adulthood

Screening for XMRV in patient PBMCs by PCR.

<p>PCR products were analyzed on agarose gels containing ethidium bromide. (A) Representative gels for non-nested <i>env</i> (top panel) and non-nested <i>gag</i> (bottom panel) PCRs are shown containing a set of three replicates for each of 5 HIV-1⁺ patient samples. A yellow arrow indicates the sole PCR band, from patient 103219, found to be comprised of XMRV DNA by sequencing. (B) A representative gel for nested <i>env</i> PCR is shown for the same 5 HIV-1⁺ patient samples depicted in (A). Vertical black arrows in (A) and (B) indicate lanes from patient 103219 containing either (A, bottom panel) a band comprised of XMRV sequence or (B) a band of the expected mobility for the target sequence. (m) 100 base pair molecular weight marker, (1∶10⁴) DNA from one infected cell diluted in DNA from 10⁴ uninfected cells used as template for positive control.</p

Detecting murine DNA by IAP PCR.

<p>PCR products were analyzed on 1.5% agarose gels containing ethidium bromide. (A) Sensitivity of the IAP PCR assay was determined by performing PCRs on titrations of EL4 murine cell line DNA in a background of 200 ng LNCaP DNA. One murine cell equivalent (1 eq) indicates 6 pg of EL4 DNA. XMRV-infected LNCaP (iLNCaP) and uninfected LNCaP (uLNCaP) were included as controls. (B) Screening results for 17 HIV-1⁺ patient samples. Arrow points to sample 103219, which tested positive for XMRV by non-nested gag PCR. (m) 100 base pair molecular weight marker, (EL4) 6 pg of murine EL4 cell line DNA without a background of human DNA.</p

Summary of XMRV screening results.

a<p>Fractions are: number of subjects scoring positive/total number of subjects screened.</p>b<p>Ab, antibody.</p

Sensitivity analysis of XMRV PCR assays.

<p>PCR products were analyzed on agarose gels containing ethidium bromide. (A) Non-nested PCR assays targeting the XMRV <i>env</i> gene (top panel) and the <i>gag</i> gene (bottom panel), and (B) a nested PCR assay targeting the XMRV <i>env</i> gene were evaluated for their ability to detect either (A) provirus in XMRV-infected PNT1A cell DNA or (B) provirus in XMRV-infected LNCaP cell DNA diluted in uninfected cell DNA. Dilutions of infected cells in uninfected cells are indicated by ratios, i.e. 1∶10⁴ indicates one infected cell diluted in 10⁴ uninfected cells. (m) 100 base pair molecular weight marker, (H₂O) water used in place of DNA template as a negative control, (u) uninfected PNT1A DNA used as template for negative control.</p

Primers used for screening PBMC DNA specimens for XMRV.

a<p>Location of 5′ end of forward primer target site to 3′ end of reverse primer target site on XMRV VP62 reference genome (accession no. DQ399707.1).</p

Birth Defects After Exposure to Efavirenz-Based Antiretroviral Therapy at Conception/First Trimester of Pregnancy: A Multicohort Analysis

Background: To investigate the association between efavirenz (EFV) use during conception or first trimester (T1) of pregnancy and the occurrence of birth defects. Setting: Seven observational studies of pregnant HIV-positive women across 13 European countries and Thailand. Methods: Individual-level data were pooled on singleton pregnancies included in participating cohorts in 2002–2015. Birth defects were coded according to ICD-10 and the EUROCAT classification. We performed mixed-effects logistic regression models to assess the association between EFV exposure in utero and likelihood of birth defects. Results: We included 24,963 live births from 21,093 women. At conception, 30.2% (7537) women were on a non–EFV-based regimen, 4.8% (1200) on EFV, and 65% (16,226) were unexposed to antiretroviral therapy (ART). There were 412 infants with ≥1 birth defect, a prevalence of 1.65% (95% confidence interval: 1.50 to 1.82). Limb/musculoskeletal and congenital heart defects were the most common defects reported. Birth defects were present in 2.4%, 1.6%, and 1.3% of infants exposed to non-EFV, EFV, and unexposed to ART during conception/T1 (P = 0.135), respectively. The association between exposure to ART during conception/T1 and birth defects remained nonsignificant in adjusted analyses, as did exposure to EFV versus non-EFV (adjusted odds ratio 0.61; 95% confidence interval: 0.36 to 1.03, P = 0.067). Among the 21 birth defects in 19 infants on EFV, no neural tube defects were reported. Conclusions: Prevalence of birth defects after exposure to EFV-based compared with non–EFV-based ART in conception/T1 was not statistically different in this multicohort study, and even lower. EFV is at least as safe as other ART drugs currently recommended for antenatal use