Nano-RECall provides an integrated pipeline for HIV-1 drug resistance testing from Oxford Nanopore sequence data.

OBJECTIVES: Low-capital-layout sequencing options from Oxford Nanopore Technologies (ONT) could assist in expanding HIV drug resistance testing to resource-limited settings. HIV drug resistance mutations often occur as mixtures, but current ONT pipelines provide a consensus sequence only. Moreover, there is no integrated pipeline that provides a drug resistance report from an ONT sequence file without intervention from skilled bioinformaticists. We therefore investigated Nano-RECall, which provides seamless drug resistance interpretation while requiring low-read coverage ONT sequence data from affordable Flongle or MinION flow cells and which provides mutation mixtures similar to Sanger Sequencing. METHODS: We compared Sanger sequencing to ONT sequencing of the same HIV-1 subtype C polymerase chain reaction (PCR) amplicons, respectively using RECall and the novel Nano-RECall bioinformatics pipelines. Amplicons were from separate assays: (a) Applied Biosystems HIV-1 Genotyping Kit (ThermoFisher) spanning protease (PR) to reverse transcriptase (RT) (PR-RT) (n = 46) and (b) homebrew integrase (IN) (n = 21). The agreement between Sanger sequences and ONT sequences was assessed at nucleotide level, and at codon level for Stanford HIV drug resistance database mutations at an optimal ONT read depth of 400 reads only. RESULTS: The average sequence similarity between ONT and Sanger sequences was 99.3% (95% CI: 99.1%-99.4%) for PR-RT and 99.6% (95% CI: 99.4%-99.7%) for INT. Drug resistance mutations did not differ for 21 IN specimens; 8 mutations were detected by both ONT- and Sanger sequencing. For the 46 PR and RT specimens, 245 mutations were detected by either ONT or Sanger, of these 238 (97.1%) were detected by both. CONCLUSIONS: The Nano-RECall pipeline, freely available as a downloadable application on a Windows computer, provides Sanger-equivalent HIV drug resistance interpretation. This novel pipeline combined with a simple workflow and multiplexing samples on ONT flow-cells would contribute to making HIV drug resistance sequencing feasible for resource-limited settings.

Cotidiano de pessoas que vivem com VIH em Bissau, Guiné-Bissau: perspectivas, desafios e vulnerabilidades / Vida diaria de las personas que viven con VIH en Bissau, Guinea-Bissau: perspectivas, desafíos y vulnerabilidades / The daily life of people living with HIV in Bissau, Guinea-Bissau: perspectives, challenges and vulnerabilities

RESUMO Objetivo compreender as perspectivas e desafios no cotidiano de pessoas após a descoberta do viver com VIH em Bissau, Guiné-Bissau, tendo em vista diferentes contextos de vulnerabilidade. Método estudo exploratório-descritivo, que utilizou entrevista semiestruturada com 16 pessoas vivendo com VIH, acompanhadas em um hospital de Bissau. Empregou-se técnica de análise de conteúdo temática. Os relatos dos participantes foram analisados a partir de duas categorias empíricas: A descoberta, os impactos e os desafios de viver com VIH; e Experiência com o antirretroviral: recomeço e perspectivas. Resultados sinalizaram que os desafios iniciam com a revelação do diagnóstico que, geralmente, desperta uma diversidade de sentimentos e comportamentos. O estigma e a discriminação estimulam a adoção do sigilo sobre o status sorológico, resultando na fragilidade de suporte emocional no enfrentamento à soropositividade. A terapia antirretroviral foi vislumbrada como esperança para o enfrentamento da doença. A vulnerabilidade social foi a dimensão que mais se destacou, e violações dos direitos humanos foram constatadas. Conclusão e implicações para a prática o estudo permite compreender as perspectivas, desafios e vulnerabilidades de pessoas que vivem com VIH. O viver com VIH merece atenção especial por parte dos profissionais de saúde que atuam no cuidado dessas pessoas, destacando-se como contribuição a relevância de um cuidado de saúde integral, em que a ética e a subjetividade estejam presentes.

RESUMEN Objetivo este estudio cualitativo tuvo como objetivo comprender perspectivas y desafíos en la vida cotidiana de las personas que viven con VIH en Bissau, Guinea-Bissau, considerando diferentes contextos de vulnerabilidad. Método estudio exploratorio-descriptivo, que utilizó una entrevista semiestructurada con 16 personas que viven con el VIH, seguido en un hospital de Bissau. Se utilizó la técnica de análisis de contenido temático. Los informes de los participantes se analizaron a partir de dos categorías empíricas: El descubrimiento, los impactos y los desafíos de vivir con el VIH; y La experiencia antirretroviral: un nuevo comienzo y perspectivas. Resultados los resultados indicaron que los desafíos comienzan con la divulgación del diagnóstico, que generalmente suscita una diversidad de sentimientos y conductas. El estigma y la discriminación fomentan la adopción del secreto sobre el estado serológico, lo que resulta en un apoyo emocional débil para hacer frente a la seropositividad. La terapia antirretroviral es una esperanza para hacer frente a la enfermedad. La vulnerabilidad social fue la dimensión que más se destacó y se encontraron violaciones a sus derechos humanos. Conclusión e implicaciones para la práctica el estudio permite comprender las perspectivas, los desafíos y las vulnerabilidades de las personas que viven con el VIH. Vivir con VIH merece especial atención por parte de los profesionales de la salud que actúan en el cuidado de estas personas, destacando como aporte la relevancia de la atención integral en salud, en la que la ética y la subjetividad están presentes.

ABSTRACT Objective to understand perspectives and challenges in the daily lives of people after the discovery of living with HIV in Bissau, Guinea-Bissau, considering different contexts of vulnerability. Method an exploratory-descriptive study conducted through semi-structured interviews with sixteen people living with HIV attending a hospital in the city of Bissau. We used the thematic content analysis technique. Participants' reports were analyzed following two empirical categories: Discovery, impacts and challenges of living with HIV; and Experience with antiretroviral therapy: new beginning and perspectives. Results the results indicate that the challenges start with the disclosure of diagnosis, which arouses a diversity of feelings and behaviors. Stigma and discrimination encourage the adoption of confidentiality about serological status, which increases the fragility of emotional support in coping with seropositivity. Antiretroviral therapy was seen as a hope for coping with the disease. Social vulnerability was the dimension that stood out the most, and human rights violations involving people living with HIV were verified. Conclusion and implications for practice the study makes it possible to understand the perspectives, challenges and vulnerabilities of people living with HIV. Living with HIV deserves special attention from health professionals who work in the care of these people, highlighting as a contribution the relevance of comprehensive health care, in which ethics and subjectivity are present.

Patient satisfaction in hemophilia: a fundamental component of medical care and an indicator of the quality of medical care provided.

Introduction: It is paramount that physicians treating people with hemophilia (PWH) assess their degree of satisfaction.Areas covered: The most important predictors of the difficulties of treatment satisfaction are the perceived barriers, the level of skills related to self-treatment, the degree of severity of hemophilia and the affective status. The use of an electronic diary improves the adherence of PWH with respect to the registration of the mandatory information of their treatment. Asymptomatic HIV-positive PWH are more dissatisfied than HIV-negative PWH due to the effects of HIV and to the physical functional disabilities caused by the disease. Regarding total knee arthroplasty (TKA) in hemophilia, it has been shown that PWH with advanced painful knee arthropathy do not achieve the same degree of satisfaction as PWH with advanced painful osteoarthritis of the knee, this being due to residual symptoms and deterioration of other joints. TKA and total hip arthroplasty (THA) are reliable surgical procedures that give PWH a high degree of satisfaction. Regarding the use of a Bio-Set reconstitution method, most PWH (82%) preferred the Bio-Set method to previous reconstitution methods.Expert opinion: Further studies on the degree of satisfaction of PWH are essential.

[Ethical challenges surrounding the ageing of HIV-positive patients]. / Enjeux éthiques autour du vieillissement des patients porteurs du VIH.

More and more HIV-positive people now belong to the 50+ age group. This ageing is accompanied by specific medical and psychosocial problems. In 2018, the French National Consultative Ethics Committee published a paper on the ethical challenges of ageing. Ageing is not easy in a society from which people living with HIV and the elderly feel excluded.

[Ageing and HIV, the experience of HIV-positive women in French-speaking Switzerland]. / Vieillissement et VIH, le vécu de femmes séropositives en Suisse romande.

Thanks to a burdensome lifelong treatment, it is now possible to lead an almost 'normal' life while being HIV-positive. The status of people living with the human immunodeficiency virus (HIV) is rarely questioned. The image of the 'dying contagious patient' has been replaced by that of the 'classic chronically ill patient'. However, their HIV-positive status still leads to discrimination. A sociological study based on the accounts of 30 HIV-positive women in French-speaking Switzerland reveals how they feel faced with the prospect of ageing with HIV.

[Living and building a life with HIV]. / Vivre et se construire avec le VIH.

In the 1980s, a young woman learns by chance that she has been HIV-positive for several years. Her world falls apart. The poor prognosis, the fear of having passed on the virus to her daughter, other people's judgements and complex treatments become a daily ordeal. Today, not only is her condition stable thanks to therapeutic advances, she also has a healthy grandson. Here, she shares her story.

["HIV-positive, but alive!"] / « Porteur du VIH, mais vivant ! ¼.

Frédéric has been HIV-positive for thirty-two years. He has had to deal with complex treatments, opportunistic diseases, the loss of his partner, indifference and rejection on the part of his friends. Yet despite everything, he has a message of hope to share: today, the virus is no longer detectable in his body. He still carries the virus but feels more alive than ever.

[Psoriasis and HIV infection, what do French dermatologists do?] / Détection et impact d'une séropositivité VIH chez les patients atteints de psoriasis. Une enquête de pratiques.

BACKGROUND: The prevalence of psoriasis among patients presenting human immunodeficiency virus (HIV)infection is the same as in the general population, but the disease is more severe and refractory, and management is more complex. The aim of this survey was to assess the practices of French dermatologists concerning both screening for HIV in psoriasis patients and therapeutic management and follow-up of psoriasis patients with HIV. PATIENTS AND METHODS: An anonymous national survey of practices was conducted by means of a questionnaire emailed to dermatologists in hospital in private practice in France between March and June 2017. RESULTS: The questionnaire was completed by 262 dermatologists. They indicated that they carried out screening for HIV in psoriasis patients presenting risk factors (79.4%), prior to biotherapy (63.4%) or different systemic treatments other than retinoids (53.1%), if the psoriasis was severe (45.8%), or in the event of worsening (37.8%). 28.7% of practitioners surveyed were in fact treating patients with psoriasis and HIV, with a mean 3.1 patients being followed up. All practitioners prescribe systemic treatment, other than cyclosporine, but they frequently consult an infectious disease specialist before prescribing these therapies. The most widely used biotherapy was etanercept (65.5% of practitioners). More intensive laboratory follow-up was conducted for these patients in 72.1% of cases. CONCLUSION: French dermatologists do not perform routine screening for HIV in psoriasis patients. However, where infection has been identified, their practices are modified accordingly. Therapeutic choices are consistent with the French recommendations. Nevertheless, recommendations appear necessary concerning HIV screening in this population.

Short Communication: Therapeutic Immunization Benefits Mucosal-Associated Invariant T Cell Recovery in Contrast to Interleukin-2, Granulocyte-Macrophage Colony-Stimulating Factor, and Recombinant Human Growth Hormone Addition in HIV-1+ Treated Patients: Individual Case Reports from Phase I Trial.

Mucosal-associated invariant T (MAIT) cell populations are reduced in frequency in HIV-1+ patients, and this disruption is associated with systemic immune activation. Reconstitution of MAIT frequency may benefit HIV-1-infected individuals; however, only recently has in vivo work been endeavored. Treatment with interleukin (IL)-2, granulocyte-macrophage colony-stimulating factor (GM-CSF), and recombinant human growth hormone (rhGH) immunotherapy combined with an HIV-1 vaccine in the context of antiretroviral therapy (ART) has shown to reconstitute CD4 T cell population numbers and function. In this study cryopreserved peripheral blood mononuclear cells (PBMCs) from 12 HIV-1+ patients who were undergoing a combination of HIV-1 vaccine and/or IL-2, GM-CSF and rhGH immunotherapy in conjunction with ART were analyzed to assess the potential of this treatment to promote MAIT cell proliferation. PBMCs were thawed from study baseline, weeks 2 and 48 time points, fluorescently stained for MAIT cell markers, and assessed by flow cytometric analysis. Matched pairs and intergroup results were statistically compared using appropriate methods. MAIT cell frequency was increased from baseline at 48 weeks in participants who received vaccine only, whereas individuals receiving IL-2, GM-CSF, and rhGH immunotherapy with or without vaccine did not show additional benefit. Although IL-2, GM-CSF, and rhGH treatment promotes CD4 T cell reconstitution and HIV-1-specific T cell function, it does not support MAIT cell recovery in patients on suppressive ART. Therapeutic immunization however has a positive effect, highlighting the importance of aiming for balanced promotion of T cell population reconstitution to impact on HIV-1 transmission and pathogenesis.

How social stigma sustains the HIV treatment gap for MSM in Mpumalanga, South Africa.

There are gaps in HIV care for men who have sex with men (MSM) in African settings, and HIV social stigma plays a significant role in sustaining these gaps. We conducted a three-year research project with 49 HIV-positive MSM in two districts in Mpumalanga Province, South Africa, to understand the factors that inform HIV care seeking behaviors. Semi-structured focus group discussions and interviews were conducted in IsiZulu, SiSwati, and some code-switching into English, and these were audio-recorded, transcribed, and translated into English. We used a constant comparison approach to analyze these data. HIV social stigma centered around gossip that sustained self-diagnosis and delayed clinical care with decisions to use traditional healers to mitigate the impact of gossip on their lives. More collaboration models are needed between traditional healers and health professionals to support the global goals for HIV testing and treatment.

Randomized controlled trial of a positive affect intervention for methamphetamine users.

BACKGROUND: Contingency management (CM) is an evidence-based intervention providing rewards in exchange for biomarkers that confirm abstinence from stimulants such as methamphetamine. We tested the efficacy of a positive affect intervention designed to boost the effectiveness of CM with HIV-positive, methamphetamine-using sexual minority men. METHODS: This attention-matched, randomized controlled trial of a positive affect intervention delivered during CM was registered on www.clinicaltrials.gov (NCT01926184). In total, 110 HIV-positive sexual minority men with biologically confirmed, recent methamphetamine use were enrolled. Five individual sessions of a positive affect intervention (n = 55) or an attention-control condition (n = 55) were delivered during three months of CM. Secondary outcomes examined over the 3-month intervention period included: 1) psychological processes relevant to affect regulation (i.e., positive affect, negative affect, and mindfulness); 2) methamphetamine craving; 3) self-reported stimulant use (past 3 months); and 4) cumulative number of urine samples that were non-reactive for stimulants (i.e., methamphetamine and cocaine) during CM. RESULTS: Those randomized to the positive affect intervention reported significant increases in positive affect during individual sessions and increases in mindfulness over the 3-month intervention period. Intervention-related improvements in these psychological processes relevant to affect regulation were paralleled by concurrent decreases in methamphetamine craving and self-reported stimulant use over the 3-month intervention period. CONCLUSIONS: Delivering a positive affect intervention may improve affect regulation as well as reduce methamphetamine craving and stimulant use during CM with HIV-positive, methamphetamine-using sexual minority men.

Potential interactions in a cohort of elderly hiv-positive patients.

OBJECTIVE: The increase of HIV-patients life expectancy leads to a new model of patient with chronic diseases and polymedicated. For this reason we ought to  know in clinical practice the prevalence of polypharmacy and drug-drug  interactions between the antiretroviral drugs and comedication in our patients in  order to identify and prevent them. METHOD: A retrospective, descriptive study carried out in > 50 years old patients on antiretroviral treatment. Results: We included 242 patients of whom 148 (61%) were receiving concomitant treatment. 243 potential interactions were detected,  where 197 are considered moderate and 46 severe, affecting 110 patients. 35  (76%) interactions were related to boosted protease inhibitors. The main  consequence of these interactions was the increase in plasma concentrations of  comedication (48%). Statins were the comedication most involved in severe  drug-druginteractions (24%), followed by inhaled corticosteroids (15%). CONCLUSIONS: Polypharmacy was found in about half of our study population and the prevalence of drug-drug interactions was high. Hospital pharmacists may  play a crucial role in their detection, management and early communication.

Objetivo: El aumento de la esperanza de vida conduce a un nuevo modelo de  paciente VIH positivo, con enfermedades crónicas y, en ocasiones, polimedicado. Pretendemos con este estudio conocer la complejidad de los tratamientos e  identificar potenciales interacciones entre antirretrovirales y medicación  domiciliaria de nuestros pacientes, con objeto de tenerlas identificadas y poder  prevenirlas.Método: Estudio descriptivo, retrospectivo, en una cohorte de pacientes con  tratamiento antirretroviral mayores de 50 años en un hospital de tercer grado. Resultados: Se incluyeron 242 pacientes, de los que 148 (61%) recibían algún  otro tratamiento. Detectamos 243 potenciales interacciones: 197 consideradas  moderadas y 46 graves; afectando a 110 pacientes. De las graves, 35 (76%) se  relacionaron con inhibidores de proteasa potenciados. La principal consecuencia  fue un aumento de las concentraciones plasmáticas del tratamiento domiciliario  (48%). Las estatinas (24%) fueron el grupo especialmente implicado en las  interacciones graves, seguidas de los corticoides inhalados (15%). Conclusiones: Prácticamente la mitad de los pacientes estaban polimedicados, observándose un elevado número de potenciales interacciones moderadas o graves. El farmacéutico de hospital debe jugar un  papel crucial en su detección, manejo y comunicación precoz.

Unplanned pregnancies and contraceptive use among HIV- positive women in care.

Among 230,360 women with diagnosed HIV in the United States (U.S.), ~ 8,500 give birth annually, and unplanned pregnancies (as with HIV-negative women) are prevalent. However, unplanned pregnancies and contraceptive use among HIV-positive women have been understudied. To examine unplanned pregnancies and contraceptive use among HIV-positive women, we used 2013-2014 data from the Medical Monitoring Project (MMP), an HIV surveillance system that produces national estimates for HIV-positive adults in care in the U.S. (Pregnancy outcome dates were from years 1986-2015 for this cohort of women who were interviewed during 2013-2014; median year of reported pregnancy outcome was year 2003). Women in HIV care and diagnosed with HIV before age 45 (reproductive age) were included. We calculated adjusted prevalence ratios (aPR) of unplanned pregnancies with 95% confidence intervals (CI). For women who were aged 18-44 years at time of interview, we computed weighted prevalences of contraceptive use (previous 12 months) by method, including permanent (i.e., sterilization), short-acting (i.e., pills, depo-progesterone acetate (DMPA)), long-acting reversible contraceptives (LARC) (i.e., implants), and barriers (i.e., condoms). Six hundred seventy-one women met criteria for the unplanned pregnancy analysis; median age at HIV diagnosis = 24.6 years, and 78.1% (CI:74.5-81.7) reported ≥ 1 unplanned pregnancy. Women reporting unplanned pregnancies were more likely to be non-Hispanic white (aPR = 1.20; CI 1.05-1.38) or non-Hispanic black (aPR = 1.14; CI 1.01-1.28) than Hispanic, to be above the poverty level (aPR = 1.09; CI 1.01-1.18), and to have not received care from an OB/GYN in the year before interview (aPR = 1.13; CI 1.04-1.22). Among 1,142 total pregnancies, 795 (69.6%) were live births; 70 (7.8%) were born HIV-positive; 42 (60%) of those born HIV-positive were unplanned pregnancies. For the contraceptives analysis (n = 957 women who were aged 18-44 at time of interview), 90.5% reported using at least one contraceptive, including 59.7% reporting barrier methods, 29.9% reporting permanent sterilization, and 22.8% reporting short-term methods in the previous year. LARC was used by only 5.3% of women. Women who reported use of LARC or DMPA were more likely to be aged 18-29 years (aPR = 3.08; CI 1.61-5.89) or 30-39 years (aPR = 2.86; CI 1.76-4.63) compared with women aged 40-44 years. Unplanned pregnancies were prevalent and LARC use was low; prevention efforts should strengthen pregnancy planning and contraceptive awareness for HIV-positive women during clinical visits.

Next wave of interventions to reduce under-five mortality in Rwanda: a cross-sectional analysis of demographic and health survey data.

BACKGROUND: Sustained investments in Rwanda's health system have led to historic reductions in under five (U5) mortality. Although Rwanda achieved an estimated 68% decrease in the national under U5 mortality rate between 2002 and 2012, according to the national census, 5.8% of children still do not reach their fifth birthday, requiring the next wave of child mortality prevention strategies. METHODS: This is a cross-sectional study of 9002 births to 6328 women age 15-49 in the 2010 Rwanda Demographic and Health Survey. We tested bivariate associations between 29 covariates and U5 mortality, retaining covariates with an odds ratio p < 0.1 for model building. We used manual backward stepwise logistic regression to identify correlates of U5 mortality in all children U5, 0-11 months, and 12-59 months. Analyses were performed in Stata v12, adjusting for complex sample design. RESULTS: Of 14 covariates associated with U5 mortality in bivariate analysis, the following remained associated with U5 mortality in multivariate analysis: household being among the poorest of the poor (OR = 1.98), child being a twin (OR = 2.40), mother having 3-4 births in the past 5 years (OR = 3.97) compared to 1-2 births, mother being HIV positive (OR = 2.27), and mother not using contraceptives (OR = 1.37) compared to using a modern method (p < 0.05 for all). Mother experiencing physical or sexual violence in the last 12 months was associated with U5 mortality in children ages 1-4 years (OR = 1.48, p < 0.05). U5 survival was associated with a preceding birth interval 25-50 months (OR = 0.67) compared to 9-24 months, and having a mosquito net (OR = 0.46) (p < 0.05 for both). CONCLUSIONS: In the past decade, Rwanda rolled out integrated management of childhood illness, near universal coverage of childhood vaccinations, a national community health worker program, and a universal health insurance scheme. Identifying factors that continue to be associated with childhood mortality supports determination of which interventions to strengthen to reduce it further. This study suggests that Rwanda's next wave of U5 mortality reduction should target programs in improving neonatal outcomes, poverty reduction, family planning, HIV services, malaria prevention, and prevention of intimate partner violence.

Peer counselling versus standard-of-care on reducing high-risk behaviours among newly diagnosed HIV-positive men who have sex with men in Beijing, China: a randomized intervention study.

INTRODUCTION: Reducing high-risk behaviours (i.e. multiple partnership, condomless anal/vaginal sex, alcohol use before sex, illicit drug use) after HIV diagnosis is critical for curtailing HIV transmission. We designed an intervention to explore peer- counselling in reducing high-risk behaviours among newly diagnosed HIV-positive Chinese men who have sex with men (MSM). METHODS: We randomized 367 newly diagnosed HIV-positive men to either standard-of-care (SOC; n = 183) or peer-counselling intervention (n = 184), and followed them for 12 months (visit at 0-, 3-, 6-, 9- and 12-month). SOC participants received counselling on high-risk behaviour reduction by clinic staff. Intervention participants received both SOC and peer counselling. A generalized estimating equation was used to compare pre-post diagnosis high-risk behaviour change; logistic regression was used to assess the likelihood of practicing high-risk behaviours between intervention and SOC participants. Both intent-to-treat and per-protocol (full-dosage) approaches were used for the analyses. RESULTS: For pre- and post-diagnosis comparisons, multiple partnership fell from 50% to 16% (p < 0.001), alcohol use before sex from 23% to 9% (p = 0.001), illicit drug use from 33% to 6% (p < 0.001), condomless anal sex from 47% to 4% (insertive from 23% to 2%; receptive from 36% to 3%; p < 0.001). In the intent-to-treat analysis accounting for repeated measures, peer counselling was more likely to reduce insertive anal sex (AOR = 0.65; 95% CI: 0.45 to 0.94), condomless anal sex (AOR = 0.27; 95% CI: 0.10 to 0.64) and illicit drug use (AOR = 0.32; 95% CI: 0.16 to 0.64). In the per-protocol analysis, peer counselling was associated with a lower likelihood of using illicit drug (OR = 0.23; 95% CI: 0.07 to 0.81) and having condomless vaginal sex with women (OR = 0.12; 95% CI: 0.07 to 0.98). CONCLUSIONS: We observed a 14 to 43% decrease in the prevalence of selected high-risk behaviours after HIV diagnosis. Peer counselling had a greater impact in reducing condomless anal sex with men, illicit drug use and condomless vaginal sex with women over time. Future studies with exclusive peer-counselling arm are necessary to test its efficacy and effectiveness among Chinese MSM. Clinical Trial Number: NCT01904877.

Development and optimization of a sensitive pseudovirus-based assay for HIV-1 neutralizing antibodies detection using A3R5 cells.

Sensitive assays for HIV-1 neutralizing antibody detection are urgently needed for vaccine immunogen optimization and identification of protective immune response levels. In this study, we developed an easy-to-use HIV-1 pseudovirus neutralization assay based on a human CD4+ lymphoblastoid cell line A3R5 by employing a high efficient pseudovirus production system. Optimal conditions for cell counts, infection time, virus dose and concentration of DEAE-dextran were tested and identified. For T-cell line-adapted tier 1 virus strains, significantly higher inhibitory efficiency was observed for both monoclonal neutralizing antibody (4 fold) and plasma (2 fold) samples in A3R5 than those in TZM-bl assay. For circulating tier 2 strains, the A3R5 pseudovirus assay showed even much higher sensitivity for both neutralizing antibody (10 fold) and plasma (9 fold) samples. When sequential neutralizing antibody seroconverting samples were tested in both A3R5 and TZM-bl assays, the seroconverting points could be detected earlier for tier 1 (15.7 weeks) and tier 2 (68.3 weeks) strains in A3R5 assay respectively. The high sensitive pseudovirus assay using more physiological target cells could serve as an alternative to the TZM-bl assay for evaluation of vaccine-induced neutralizing antibodies and identification of the correlates of protection.

Sequential Vacc-4x and romidepsin during combination antiretroviral therapy (cART): Immune responses to Vacc-4x regions on p24 and changes in HIV reservoirs.

OBJECTIVES: The REDUC clinical study Part B investigated Vacc-4x/rhuGM-CSF therapeutic vaccination prior to HIV latency reversal using romidepsin. The main finding was a statistically significant reduction from baseline in viral reservoir measurements. Here we evaluated HIV-specific functional T-cell responses following Vacc-4x/rhuGM-CSF immunotherapy in relation to virological outcomes on the HIV reservoir. METHODS: This study, conducted in Aarhus, Denmark, enrolled participants (n = 20) with CD4>500 cells/mm3 on cART. Six Vacc-4x (1.2 mg) intradermal immunizations using rhuGM-CSF (60 µg) as adjuvant were followed by 3 weekly intravenous infusions of romidepsin (5 mg/m2). Immune responses were determined by IFN-γ ELISpot, T-cell proliferation to p24 15-mer peptides covering the Vacc-4x region, intracellular cytokine staining (ICS) to the entire HIVGag and viral inhibition. RESULTS: The frequency of participants with CD8+ T-cell proliferation assay positivity was 8/16 (50%) at baseline, 11/15 (73%) post-vaccination, 6/14 (43%) during romidepsin, and 9/15 (60%)post-romidepsin. Participants with CD8+ T-cell proliferation assay positivity post-vaccination showed reductions in total HIV DNA post-vaccination (p = 0.006; q = 0.183), post-latency reversal (p = 0.005; q = 0.183), and CA-RNA reductions post-vaccination (p = 0.015; q = 0.254). Participants (40%) were defined as proliferation 'Responders' having ≥2-fold increase in assay positivity post-baseline. Robust ELISpot baseline responses were found in 87.5% participants. No significant changes were observed in the proportion of polyfunctional CD8+ T-cells to HIVGag by ICS. There was a trend towards increased viral inhibition from baseline to post-vaccination (p = 0.08). CONCLUSIONS: In this 'shock and kill' approach supported by therapeutic vaccination, CD8+ T-cell proliferation represents a valuable means to monitor functional immune responses as part of the path towards functional HIV cure.

Recent Recommendations for Management of Human Immunodeficiency Virus-Positive Patients.

Human immunodeficiency virus (HIV) infection has become a chronic condition. HIV is not a valid reason to deny, delay, or withhold dental treatment. There are no absolute contraindications and few complications associated with comprehensive oral health care treatment delivered in an outpatient setting for asymptomatic HIV-infected patients and clinically stable patients with AIDS. Consultation with the patient's medical provider and modifications in the delivery of dental treatment may be necessary when treating patients with advanced HIV disease or other comorbid conditions. Oral health care is an integral and important part of comprehensive health care for all patients with HIV/AIDS.

How acceptable is it for HIV positive African, Caribbean and Black women to provide breast milk/fluid samples for research purposes?

BACKGROUND: The African, Caribbean and Black communities have been found to be reluctant to participate in health research in North America. This is partly attributed to historical experiences as well as their cultural beliefs. Cultural beliefs about the uses of breast milk/fluids could further hinder the participation of African, Caribbean, and Black communities in research involving the collection of breast milk/fluids samples. METHODS: We conducted 17 in-depth interviews and three group interviews (n = 10) with HIV+ African, Caribbean and Black women living in Ontario, Canada to explore their cultural beliefs about breast milk/fluids and their acceptance of participating in research that involves the provision of breast fluid samples. STUDY DESIGN: Qualitative study involving in-depth interviews. RESULTS: Our respondents believed that breast milk/fluids should be used for infant feeding and for curative purposes for a variety of children's health ailments as well as ailments experienced by other family members. The cultural belief that breast milk/fluids could be used to bewitch the baby and mother and the perception that it is intrusive (equating breast milk/fluids research to DNA testing), could prevent African, Caribbean and Black women from participating in research involving the collection of breast milk/fluids. Despite these fears, some respondents expressed that they would participate if the research results would benefit them directly, for example, by finding a cure for HIV, enabling HIV+ mothers to breastfeed, or contributing to developing new drugs or vaccines for HIV. Women's recommendations to facilitate successful recruitment included giving incentives to participants, and employing a recruiter who was trustworthy, informed, and culturally sensitive. CONCLUSION: Cultural beliefs could present barriers to recruitment and participation of Africa, Caribbean and Black communities in health research involving breast milk/fluid samples. Successful recruitment for future studies would necessitate researchers to be culturally aware of the beliefs held by African, Caribbean and Black women, to build trust, and use an appropriate recruiter. While the findings relate to breast milk/fluids, the suggested recommendations for facilitating recruitment of research participants from these communities may be useful to consider when recruiting ethnically and culturally similar participants for research involving biological samples.