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.