A Double-Blind Randomized Phase I Clinical Trial Targeting ALVAC-HIV Vaccine to Human Dendritic Cells

BACKGROUND: We conducted a novel pilot study comparing different delivery routes of ALVAC-HIV (vCP205), a canarypox vaccine containing HIV gene inserts: env, gag and pol. We explored the concept that direct ex vivo targeting of human dendritic cells (DC) would enhance the immune response compared to either conventional intramuscular or intradermal injections of the vaccine alone. METHODOLOGY/PRINCIPAL FINDINGS: Healthy HIV-1 uninfected volunteers were administered ALVAC-HIV or placebo by intramuscular injection (i.m.), intradermal injection (i.d.) or subcutaneous injection (s.q.) of autologous ex vivo transfected DC at months 0, 1, 3 and 6. All vaccine delivery routes were well tolerated. Binding antibodies were observed to both the ALVAC vector and HIV-1 gp160 proteins. Modest cellular responses were observed in 2/7 individuals in the DC arm and 1/8 in the i.m. arm as determined by IFN-γ ELISPOT. Proliferative responses were most frequent in the DC arm where 4/7 individuals had measurable responses to multiple HIV-1 antigens. Loading DC after maturation resulted in lower gene expression, but overall better responses to both HIV-1 and control antigens, and were associated with better IL-2, TNF-α and IFN-γ production. CONCLUSIONS/SIGNIFICANCE: ALVAC-HIV delivered i.m., i.d. or s.q. with autologous ex vivo transfected DC proved to be safe. The DC arm was most immunogenic. Proliferative immune responses were readily detected with only modest cytotoxic CD8 T cell responses. Loading mature DC with the live viral vaccine induced stronger immune responses than loading immature DC, despite increased transgene expression with the latter approach. Volunteers who received the autologous vaccine loaded mature DC developed a broader and durable immune response compared to those vaccinated by conventional routes. TRIAL REGISTRATION: ClinicalTrials.gov NCT00013572

The malleability of stigmatizing attitudes: Combining imagined social contact with implicit attitude feedback

© 2016, Copyright © Taylor & Francis Group, LLC. Research is reported that examines whether imagined social contact combined with implicit attitude feedback may be an effective intervention for inducing changes in attitudes toward mental ill health. The Implicit Relational Assessment Procedure (IRAP) captured participants’ implicit attitudes toward individuals with a mental illness and provided a measure of attitude bias. Forty-eight participants (17 male, 95.8% White British) were randomly assigned to one of four experimental conditions: (1) Imagined social contact with implicit attitude feedback, (2) imagined social contact without feedback, (3) control with feedback, and (4) control without feedback. This resulted in a data set detailing 12,288 implicit responses, with each participant completing 256 IRAP trials. Participants then completed an attitude change assessment 24 hours later. Results revealed that imagined social contact was successful in changing implicit attitudes, with the addition of implicit attitude feedback further strengthening participants’ positive attitudes toward mental ill health. Explicit attitudes remained unaffected. These findings are the first to highlight the practical importance of combining imagined social contact with implicit attitude feedback to improve attitudes toward out-groups who are stigmatized

A Highly Intensified ART Regimen Induces Long-Term Viral Suppression and Restriction of the Viral Reservoir in a Simian AIDS Model

Stably suppressed viremia during ART is essential for establishing reliable simian models for HIV/AIDS. We tested the efficacy of a multidrug ART (highly intensified ART) in a wide range of viremic conditions (103–107 viral RNA copies/mL) in SIVmac251-infected rhesus macaques, and its impact on the viral reservoir. Eleven macaques in the pre-AIDS stage of the disease were treated with a multidrug combination (highly intensified ART) consisting of two nucleosidic/nucleotidic reverse transcriptase inhibitors (emtricitabine and tenofovir), an integrase inhibitor (raltegravir), a protease inhibitor (ritonavir-boosted darunavir) and the CCR5 blocker maraviroc. All animals stably displayed viral loads below the limit of detection of the assay (i.e. <40 RNA copies/mL) after starting highly intensified ART. By increasing the sensitivity of the assay to 3 RNA copies/mL, viral load was still below the limit of detection in all subjects tested. Importantly, viral DNA resulted below the assay detection limit (<2 copies of DNA/5*105 cells) in PBMCs and rectal biopsies of all animals at the end of the follow-up, and in lymph node biopsies from the majority of the study subjects. Moreover, highly intensified ART decreased central/transitional memory, effector memory and activated (HLA-DR+) effector memory CD4+ T-cells in vivo, in line with the role of these subsets as the main cell subpopulations harbouring the virus. Finally, treatment with highly intensified ART at viral load rebound following suspension of a previous anti-reservoir therapy eventually improved the spontaneous containment of viral load following suspension of the second therapeutic cycle, thus leading to a persistent suppression of viremia in the absence of ART. In conclusion, we show, for the first time, complete suppression of viral load by highly intensified ART and a likely associated restriction of the viral reservoir in the macaque AIDS model, making it a useful platform for testing potential cures for AIDS

Recommendations for Enhancing Psychosocial Support of NICU Parents through Staff Education and Support

Providing psychosocial support to parents whose infants are hospitalized in the neonatal intensive care unit (NICU) can improve parents’ functioning as well as their relationships with their babies. Yet, few NICUs offer staff education that teaches optimal methods of communication with parents in distress. Limited staff education in how to best provide psychosocial support to families is one factor that may render those who work in the NICU at risk for burnout, compassion fatigue and secondary traumatic stress syndrome. Staff who develop burnout may have further reduced ability to provide effective support to parents and babies. Recommendations for providing NICU staff with education and support are discussed. The goal is to deliver care that exemplifies the belief that providing psychosocial care and support to the family is equal in importance to providing medical care and developmental support to the baby

T-cell exhaustion, co-stimulation and clinical outcome in autoimmunity and infection.

The clinical course of autoimmune and infectious disease varies greatly, even between individuals with the same condition. An understanding of the molecular basis for this heterogeneity could lead to significant improvements in both monitoring and treatment. During chronic infection the process of T-cell exhaustion inhibits the immune response, facilitating viral persistence. Here we show that a transcriptional signature reflecting CD8 T-cell exhaustion is associated with poor clearance of chronic viral infection, but conversely predicts better prognosis in multiple autoimmune diseases. The development of CD8 T-cell exhaustion during chronic infection is driven both by persistence of antigen and by a lack of accessory 'help' signals. In autoimmunity, we find that where evidence of CD4 T-cell co-stimulation is pronounced, that of CD8 T-cell exhaustion is reduced. We can reproduce the exhaustion signature by modifying the balance of persistent stimulation of T-cell antigen receptors and specific CD2-induced co-stimulation provided to human CD8 T cells in vitro, suggesting that each process plays a role in dictating outcome in autoimmune disease. The 'non-exhausted' T-cell state driven by CD2-induced co-stimulation is reduced by signals through the exhaustion-associated inhibitory receptor PD-1, suggesting that induction of exhaustion may be a therapeutic strategy in autoimmune and inflammatory disease. Using expression of optimal surrogate markers of co-stimulation/exhaustion signatures in independent data sets, we confirm an association with good clinical outcome or response to therapy in infection (hepatitis C virus) and vaccination (yellow fever, malaria, influenza), but poor outcome in autoimmune and inflammatory disease (type 1 diabetes, anti-neutrophil cytoplasmic antibody-associated vasculitis, systemic lupus erythematosus, idiopathic pulmonary fibrosis and dengue haemorrhagic fever). Thus, T-cell exhaustion plays a central role in determining outcome in autoimmune disease and targeted manipulation of this process could lead to new therapeutic opportunities

Determinations of Levels of Human Immunodeficiency Virus Type 1 RNA in Plasma: Reassessment of Parameters Affecting Assay Outcome

Appropriate interpretation of HIV-1 RNA levels requires an understanding of differences in test results due to multiple factors, which include assay and biological variation as well as specimen-handling conditions. Multiple investigations with diverse patient populations and assays have suggested that the contributions of technical and biological variations to RNA levels were quite consistent and predictable and in the range of 0.3 to 0.6 log10 RNA copies/ml. To date, all of the studies that have assessed variations in the levels of HIV-1 RNA measured have been limited primarily to isolates of the B clade; thus, what is lacking is knowledge of the degree to which the clade subtype influences assay variation and whether the biological variation observed with the clade B subtype is consistent for other clades. The major finding from the workshop was the unexpected stability of the HIV-1 RNA collected and stored under a variety of specimen handling conditions. HIV-1 RNA was shown to be relatively stable in whole blood, plasma, and serum, with the greatest stability being in plasma. Separated plasma was found to have stable titers even after storage at room temperature for 24 to 48 h and repeated freeze-thaw cycles. Within the constraints of the studies described here, the potential differences in RNA levels due to various specimen- handling conditions were not large (10 to 20% due to the anticoagulant type used in the collection tube [30 to 80% if serum rather than plasma is used], 10 to 30% due to time at RT prior to processing within 24 h, 30 to 80% due to the use of a storage temperature of -20 or -80°C). Thus, the anticipated RNA levels for nonideally collected and processed plasma specimens may be only about 130% (0.11 log10) less than those for plasma specimens collected and processed ideally (assuming that these differences are additive). This 130% difference is relatively small compared to the potential total average standard deviation of up to about 400% or 0.6 log10 RNA copies/ml due to intra- and interassay (both 0.1 to 0.2 log10) and biological (0.1 to 0.2 log10) RNA copies/ml factors. On the basis of these findings, workshop participants concluded that retrospective studies, including those which have used sera or heparinized samples, should show biological comparability to studies performed under ideal conditions, and thus both retrospective and prospective studies are useful in providing an understanding of the role of HIV-1 RNA levels in blood in transmission and disease progression. However, for prospectively designed studies, workshop participants recommended that blood for quantitative HIV-1 RNA testing ideally be collected in tubes containing EDTA, processed within 6 h of collection (but up to 24 h is still acceptable), and then stored at -80°C until assayed. Novel methodological approaches which could be useful in diagnosing and quantitating viral load in developing countries were also described, i.e., the use of DPSs, or in other body fluids such as cervical-vaginal secretions, i.e., sno-strip wicks. Finally, workshop participants determined what laboratory evaluations, including assays of HIV-1 RNA levels, with blood samples should be a priority in pediatric cohort studies while acknowledging that this ultimately depends on the study question being asked. Recommendations concerning specimen handling were then developed for international and domestic studies that use assays for detection of HIV-1 RNA. The findings reported herein underscore the continued need for the exchange of information among investigation and industry with the aim of elucidating the technological parameters that influence the assays used to evaluate HIV-1 disease and therapeutic interventions. Only by understanding the factors that affect assay outcome can we appropriately discern their value and use in clinical studies and for patient management

Nurse forecasting in Europe (RN4CAST): Rationale, design and methodology

Contains fulltext : 97171.pdf (postprint version ) (Open Access)BACKGROUND: Current human resources planning models in nursing are unreliable and ineffective as they consider volumes, but ignore effects on quality in patient care. The project RN4CAST aims innovative forecasting methods by addressing not only volumes, but quality of nursing staff as well as quality of patient care. METHODS/DESIGN: A multi-country, multilevel cross-sectional design is used to obtain important unmeasured factors in forecasting models including how features of hospital work environments impact on nurse recruitment, retention and patient outcomes. In each of the 12 participating European countries, at least 30 general acute hospitals were sampled. Data are gathered via four data sources (nurse, patient and organizational surveys and via routinely collected hospital discharge data). All staff nurses of a random selection of medical and surgical units (at least 2 per hospital) were surveyed. The nurse survey has the purpose to measure the experiences of nurses on their job (e.g. job satisfaction, burnout) as well as to allow the creation of aggregated hospital level measures of staffing and working conditions. The patient survey is organized in a sub-sample of countries and hospitals using a one-day census approach to measure the patient experiences with medical and nursing care. In addition to conducting a patient survey, hospital discharge abstract datasets will be used to calculate additional patient outcomes like in-hospital mortality and failure-to-rescue. Via the organizational survey, information about the organizational profile (e.g. bed size, types of technology available, teaching status) is collected to control the analyses for institutional differences.This information will be linked via common identifiers and the relationships between different aspects of the nursing work environment and patient and nurse outcomes will be studied by using multilevel regression type analyses. These results will be used to simulate the impact of changing different aspects of the nursing work environment on quality of care and satisfaction of the nursing workforce. DISCUSSION: RN4CAST is one of the largest nurse workforce studies ever conducted in Europe, will add to accuracy of forecasting models and generate new approaches to more effective management of nursing resources in Europe

Comparative Expression Profile of miRNA and mRNA in Primary Peripheral Blood Mononuclear Cells Infected with Human Immunodeficiency Virus (HIV-1)

Host cells respond to exogenous infectious agents such as viruses, including HIV-1. Studies have evaluated the changes associated with virus infection at the transcriptional and translational levels of the cellular genes involved in specific pathways. While this approach is useful, in our view it provides only a partial view of genome-wide changes. Recently, technological advances in the expression profiling at the microRNA (miRNA) and mRNA levels have made it possible to evaluate the changes in the components of multiple pathways. To understand the role of miRNA and its interplay with host cellular gene expression (mRNA) during HIV-1 infection, we performed a comparative global miRNA and mRNA microarray using human PBMCs infected with HIV-1. The PBMCs were derived from multiple donors and were infected with virus generated from the molecular clone pNL4-3. The results showed that HIV-1 infection led to altered regulation of 21 miRNAs and 444 mRNA more than 2-fold, with a statistical significance of p<0.05. Furthermore, the differentially regulated miRNA and mRNA were shown to be associated with host cellular pathways involved in cell cycle/proliferation, apoptosis, T-cell signaling, and immune activation. We also observed a number of inverse correlations of miRNA and mRNA expression in infected PBMCs, further confirming the interrelationship between miRNA and mRNA regulation during HIV-1 infection. These results for the first time provide evidence that the miRNA profile could be an early indicator of host cellular dysfunction induced by HIV-1