Supporting Data for "Reversing B Cell Exhaustion in Chronic HIV-1 Infection by Targeting the Δ42PD-1-AKT1/FOXO1 Signaling Axis"

1. Dataset OverviewAbstract: This dataset comprises experimental data derived from peripheral blood mononuclear cells (PBMCs). A total of 165 PBMC samples were obtained from 162 individuals living with HIV (PLWH) and 36 HIV-1 seronegative healthy controls, matched for age and sex. The dataset encompasses a series of in vitro functional and phenotypic assays performed on these samples. The experimental objectives included: CFSE staining to track lymphocyte proliferation dynamics; multiparameter flow cytometry to phenotype immune cell subsets and compare profiles between PLWH and controls; and functional assays involving antibody stimulation to observe potential therapeutic effects on immune responses.2. Methodology and Data CollectionStudy Design: The investigation followed a sequential experimental pipeline: 1) Phenotypic Characterization: Initial immunophenotyping of donor samples. 2) Dynamic Functional Analysis: Assessment of cellular proliferation and response kinetics. 3) Pathway Exploration: Investigation of signaling and metabolic pathways. 4) Therapeutic Effect Observation: Evaluation of in vitro responses to antibody-based treatments.Data Collection Tools: Primary data were generated using Fluorescence-Activated Cell Sorting (FACS) analyzers (e.g., BD Aria III) and associated software (e.g., BD FACSDiva).Data Collection Workflow:PBMCs were isolated via density gradient centrifugation (Ficoll-Paque).For proliferation: Cells were stained with CFSE and cultured under specific conditions before flow cytometric analysis.For phenotyping: Cells were stained with fluorochrome-conjugated antibodies against target markers (e.g., CD19, PD-1，Δ42PD-1) and acquired on the flow cytometer.For functional assays: Cells were treated with specific antibodies (e.g., CH34) and subsequently analyzed for changes in proliferation, activation markers, or cytokine production.Raw flow cytometry (.fcs) and image files were processed and analyzed in GraphPad Prism (versions 9/10). Data were organized and statistically analyzed according to specific hypotheses using tests such as unpaired t-tests, Mann-Whitney U tests, ANOVA, or linear mixed-effects models, as appropriate.Data Encoding: All donor samples are identified by a unique numerical code. A separate, password-protected master key links these codes to de-identified donor metadata (e.g., group classification: age, sex, clinical parameters). No personally identifiable information is present in the experimental dataset.File Format: The final analyzed datasets and graph layouts are provided in .pzfx (Prism project) and .prism (Prism template) formats, ensuring full compatibility with GraphPad Prism software for direct access to analysis steps and raw data plots.3. Ethical and Legal Compliance StatementEthical Approval: This study was conducted in full compliance with the ethical principles outlined in the Declaration of Helsinki. The study protocol was reviewed and approved by multiple independent ethics committees:The Institutional Review Board of the University of Hong Kong/Hospital Authority Hong Kong West Cluster (Reference: UW 19-834).The Hong Kong East Cluster Research Ethics Committee.The Kowloon West Cluster Research Ethics Committee.The Ethics Committee of the Shenzhen Third People's Hospital (Approval No.: 2018080).Informed Consent: Written informed consent was obtained from all participants prior to their enrollment in the study and sample collection.Privacy and Confidentiality:All data are de-identified. Donors are referenced only by the numerical code described above.The master linking key is stored separately in an encrypted file with restricted access.All original data files (e.g., .fcs, images) and analysis files are stored on a secure, password-protected server with access limited to authorized members of the research team.4. Availability StatementRepository: This dataset is permanently archived and hosted on the [HKU DATAHUB] repository.Identifier: DOI:10.25442/hku.30790532.License: CC BY-NC 4.0.