Protocol for measuring killing capacity and intracellular cytokine production of human HIV antigen-specific CD8 T cells using flow cytometry.

Here, we present a protocol to evaluate the killing capacity and functional profile of human HIV-specific CD8 T cells. We describe steps for culturing peripheral blood mononuclear cells (PBMCs) from patients with HIV on antiretroviral therapy (ART) with HIV peptides ex vivo and quantifying HIV-specific CD8 T cell killing using flow cytometry. We then detail procedures for integrating the established killing assay with intracellular cytokine staining (ICS) and assessing CD8 T cell function.

GITR activation impairs CD8 T cell function in people with HIV on antiretroviral therapy.

Glucocorticoid-induced tumor necrosis factor related protein (GITR) is a co-stimulatory immune checkpoint molecule constitutively expressed on regulatory T cells (T) and on activated T conventional cells (T). In blood collected from PWH on suppressive ART, GITR expression was reduced in multiple activated CD4 and CD8 T cell subsets but was increased in Tregs. HIV specific CD8 T cells expressed higher levels of GITR and programmed cell death protein 1 (PD-1) compared to total CD8 T cells.

Assessing the Fidelity of an Affirmative Cognitive Behavioral Group Intervention.

Purpose: Support implementation fidelity in intervention research with lesbian, gay, bisexual, transgender, queer, and sexual and gender diverse (LGBTQ+) populations, this study explores the systematic development of a fidelity process for AFFIRM, an evidence-based, affirmative cognitive behavioral therapy group intervention for LGBTQ+ youth and adults.

Method: As part of a clinical trial, the AFFIRM fidelity checklist was designed to assess clinician adherence. A total of 151 audio-recorded group sessions were coded by four trained raters.

Memory CD4 T cells that co-express PD1 and CTLA4 have reduced response to activating stimuli facilitating HIV latency.

Programmed cell death 1 (PD1) and cytotoxic T lymphocyte-associated protein 4 (CTLA4) suppress CD4 T cell activation and may promote latent HIV infection. By performing leukapheresis (n = 21) and lymph node biopsies (n = 8) in people with HIV on antiretroviral therapy (ART) and sorting memory CD4 T cells into subsets based on PD1/CTLA4 expression, we investigate the role of PD1 and CTLA 4 in HIV persistence. We show that double-positive (PD1CTLA4) cells in blood contain more HIV DNA compared with double-negative (PD1CTLA4) cells but still have a lower proportion of cells producing multiply spliced HIV RNA after stimulation as well as reduced upregulation of T cell activation and proliferation markers.

Demonstrating LGBTQ+ Affirmative Practice in Groups:: Developing Competence through Simulation-Based Learning.

Lesbian, gay, bisexual, transgender, queer, and other sexual and/or gender minority (LGBTQ+) populations experience significant mental and behavioral health disparities. Social workers are uniquely positioned to address these vulnerabilities. However, clinical graduate education has not effectively promoted or taught competent practice with LGBTQ+ populations.

Counter-Narratives of Structural Oppressions, Stigma and Resistance, and Reproductive and Sexual Health Among Youth Experiencing Homelessness.

Youth experiencing homelessness (YEH) face myriad injustices regarding their reproductive and sexual health and rights. Reproductive and sexual health research with YEH often explores condom-use and sexually transmitted infections, potentially contributing to narrow conceptualizations of YEH as "unclean" or in need of disease-screening. A narrative theory perspective was applied to this study, which allowed for the emergence of alternative storylines, or counter-narratives, which attend to manifestations of power and oppression within the lives of marginalized individuals.

An Inspiring Journey of Hope and Persistence: Life Lessons with Françoise Barré-Sinoussi.

Three early-career female virologists sat down with a distinguished Nobel laureate to discuss two pandemics, 39 years apart [...

Multiparameter immunohistochemistry analysis of HIV DNA, RNA and immune checkpoints in lymph node tissue.

The main barrier to a cure for HIV is the persistence of long-lived and proliferating latently infected CD4 T-cells despite antiretroviral therapy (ART). Latency is well characterized in multiple CD4 T-cell subsets, however, the contribution of regulatory T-cells (Tregs) expressing FoxP3 as well as immune checkpoints (ICs) PD-1 and CTLA-4 as targets for productive and latent HIV infection in people living with HIV on suppressive ART is less well defined. We used multiplex detection of HIV DNA and RNA with immunohistochemistry (mIHC) on formalin-fixed paraffin embedded (FFPE) cells to simultaneously detect HIV RNA and DNA and cellular markers.

Combination Immune Checkpoint Blockade Enhances IL-2 and CD107a Production from HIV-Specific T Cells Ex Vivo in People Living with HIV on Antiretroviral Therapy.

In people with HIV (PWH) on antiretroviral therapy (ART), immune dysfunction persists, including elevated expression of immune checkpoint (IC) proteins on total and HIV-specific T cells. Reversing immune exhaustion is one strategy to enhance the elimination of HIV-infected cells that persist in PWH on ART. We aimed to evaluate whether blocking CTL-associated protein 4 (CTLA-4), programmed cell death protein 1 (PD-1), T cell Ig domain and mucin domain 3 (TIM-3), T cell Ig and ITIM domain (TIGIT) and lymphocyte activation gene-3 (LAG-3) alone or in combination would enhance HIV-specific CD4 and CD8 T cell function ex vivo.

Cell-Associated Human Immunodeficiency Virus (HIV) Ribonucleic Acid Has a Circadian Cycle in Males With HIV on Antiretroviral Therapy.

Background: Circadian transcription factors that regulate cell-autonomous circadian clocks can also increase human immunodeficiency virus (HIV) transcription in vitro. We aimed to determine whether circadian variation in HIV transcription exists in people with HIV (PWH) on antiretroviral therapy (ART).

Methods: We performed a prospective observational study of male PWH on ART, sampling blood every 4 hours for 24 hours.

Towards an integrative self: a digital photo elicitation study of resilience among key marginalized populations of sexual and gender minority youth.

Sexual and gender minority youth (SGMY) experience unique challenges related to identity and disclosure, and cope in vibrant ways. Qualitative research has not yet fulsomely explored the risk, resilience, and identity intersections that impact vulnerable SGMY wellbeing. Methods: This digital photo-elicitation study (QueerView) recruited thirty SGMY (aged 14-29) from priority populations that had one or more of the following experiences:  trans and gender diverse, homelessness, child welfare, and immigration.

Efficacy of affirmative cognitive behavioural group therapy for sexual and gender minority adolescents and young adults in community settings in Ontario, Canada.

Objective: This study tested the efficacy of AFFIRM, a brief affirmative cognitive-behavioural group intervention tailored to reduce psychosocial distress and improve coping among sexual and gender minority adolescents and young adults (SGMY).

Method: SGMY (n = 138; M age = 22.44) were allocated to immediate 8-week AFFIRM intervention delivered at 12 community-based organisations or an 8-week waitlisted control.

Critical reflections and reflexivity on responding to the needs of LGBTQ+ youth in a global pandemic.

The global community has been significantly impacted by the COVID-19 global pandemic. LGBTQ+ (i.e.

Adapting Clinical Skills to Telehealth: Applications of Affirmative Cognitive-Behavioral Therapy with LGBTQ+ Youth.

Online social work services (e.g., telemental health; telebehavioral health; virtual care; telehealth) present significant opportunities for clinical social workers to provide effective care to marginalized populations, such as LGBTQ+ youth.

Multiply spliced HIV RNA is a predictive measure of virus production ex vivo and in vivo following reversal of HIV latency.

Background: One strategy being pursued to clear latently infected cells that persist in people living with HIV (PLWH) on antiretroviral therapy (ART) is to activate latent HIV infection with a latency reversing agent (LRA). Surrogate markers that accurately measure virus production following an LRA are needed.

Methods: We quantified cell-associated unspliced (US), multiply spliced (MS) and supernatant (SN) HIV RNA by qPCR from total and resting CD4+ T cells isolated from seven PLWH on ART before and after treatment ex vivo with different LRAs, including histone deacetylase inhibitors (HDACi).

AFFIRM Online: Utilising an Affirmative Cognitive-Behavioural Digital Intervention to Improve Mental Health, Access, and Engagement among LGBTQA+ Youth and Young Adults.

Digital mental health interventions may enable access to care for LGBTQA+ youth and young adults that face significant threats to their wellbeing. This study describes the preliminary efficacy of AFFIRM Online, an eight-session manualised affirmative cognitive behavioural group intervention delivered synchronously. Participants (M = 21.

QueerVIEW: Protocol for a Technology-Mediated Qualitative Photo Elicitation Study With Sexual and Gender Minority Youth in Ontario, Canada.

Background: The experiences of resilience and intersectionality in the lives of contemporary sexual and gender minority youth (SGMY) are important to explore. SGMY face unique experiences of discrimination in both online and offline environments, yet simultaneously build community and seek support in innovative ways. SGMY who identify as transgender, trans, or gender nonconforming and have experiences with child welfare, homelessness, or immigration have been particularly understudied.

Diverse effects of interferon alpha on the establishment and reversal of HIV latency.

HIV latency is the major barrier to a cure for people living with HIV (PLWH) on antiretroviral therapy (ART) because the virus persists in long-lived non-proliferating and proliferating latently infected CD4+ T cells. Latently infected CD4+ T cells do not express viral proteins and are therefore not visible to immune mediated clearance. Therefore, identifying interventions that can reverse latency and also enhance immune mediated clearance is of high interest.

Combination Immune Checkpoint Blockade to Reverse HIV Latency.

In people living with HIV on antiretroviral therapy, HIV latency is the major barrier to a cure. HIV persists preferentially in CD4 T cells expressing multiple immune checkpoint (IC) molecules, including programmed death (PD)-1, T cell Ig and mucin domain-containing protein 3 (TIM-3), lymphocyte associated gene 3 (LAG-3), and T cell immunoreceptor with Ig and ITIM domains (TIGIT). We aimed to determine whether these and other IC molecules have a functional role in maintaining HIV latency and whether blocking IC molecules with Abs reverses HIV latency.

Prevalence of frailty in a tertiary hospital: A point prevalence observational study.

Introduction: Frailty is an important concept in modern healthcare due to its association with adverse outcomes. Its prevalence varies in the literature and there is a paucity of literature looking at the prevalence of frailty in an inpatient setting. Its significance lies on its impact on resource utilisation and costs.

Myeloid Dendritic Cells Induce HIV Latency in Proliferating CD4 T Cells.

HIV latency occurs predominantly in long-lived resting CD4 T cells; however, latent infection also occurs in T cell subsets, including proliferating CD4 T cells. We compared the establishment and maintenance of latent infection in nonproliferating and proliferating human CD4 T cells cocultured with syngeneic myeloid dendritic cells (mDC). Resting CD4 T cells were labeled with the proliferation dye eFluor 670 and cultured alone or with mDC, plasmacytoid dendritic cells, or monocytes in the presence of staphylococcal enterotoxin B (SEB).

Meeting Management Standards and Improvement in Clinical Outcomes Among Patients With Hip Fractures.

Objectives: To determine the association between management standards and clinical outcomes among patients with hip fracture (HF).

Methods: Data from a prospective cohort study were linked with hospital administration data.

Results: In 2014 and 2015, 493 patients had surgery for HF.

Validity of routinely collected data in identifying hip fractures at a major tertiary hospital in Australia.

Objectives: To examine the validity of routinely collected data in identifying hip fractures (HFs) and to identify factors associated with incorrect coding.

Method: In a prospective cohort study between January 2014 and June 2016, HFs were identified using physician diagnosis and diagnostic imaging and were recorded in a Registry. Records of HFs in the health information exchange (HIE) were identified using International Statistical Classification of Diseases and Related Health Problems, Tenth Revision, Australian Modification/Australian Classification of Health Interventions/Australian Coding Standards codes.

Elective surgical outcomes of patients in Christchurch, New Zealand.

Aim: Most studies assessing mortality after surgery have been undertaken in major public hospitals or are procedure specific. The aim of this study was to determine mortality after elective surgery at a total community level with inclusion of all patients undergoing elective surgery.

Method: This was a prospective study of all patients that underwent elective surgery in Christchurch, New Zealand, within a calendar month.