PBMC Activation and RNA Isolation

Freshly thawed PBMCs from PWH were counted. Half of the PBMCs were cultured in complete RPMI media alone while the remaining PBMCs were activated by incubation in the presence of anti-CD3/CD28 beads added at the ratio of 1 bead:1 cell for 3 days. To prevent the spread of infection, 1 μM of Efavirenz (EFV) and 1 μM of Saquinavir (SQV) were also added. Activated as well as unstimulated PBMCs were collected at the same time, and RNA isolation was performed using a Qiagen AllPrep DNA/RNA mini kit.

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Basukala B., Rossi S., Bendiks S., Gnatienko N., Patts G., Krupitsky E., Lioznov D., So-Armah K., Sagar M., Cheng C, & Henderson A.J. (2023). Virally Suppressed People Living with HIV Who Use Opioids Have Diminished Latency Reversal. Viruses, 15(2), 415.

Publication 2023

AllPrep DNA/RNA mini kit

Anti cd3 Cell Cultured media Efavirenz Infection Isolation Saquinavir

Corresponding Organization : Boston University

Other organizations : Boston Medical Center, First Pavlov State Medical University of St. Petersburg, St.Petersburg V.M.Bekhterev Psychoneurological Research Institute, University of California, San Diego

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Variable analysis

independent variables

Activation of PBMCs: PBMCs were either cultured in complete RPMI media alone or activated by incubation in the presence of anti-CD3/CD28 beads

dependent variables

RNA isolation: RNA was isolated from both activated and unstimulated PBMCs

control variables

Cell type: Freshly thawed PBMCs from PWH (people with HIV) were used
Anti-retroviral drugs: 1 μM of Efavirenz (EFV) and 1 μM of Saquinavir (SQV) were added to prevent the spread of infection

controls

Positive control: Activated PBMCs
Negative control: Unstimulated PBMCs

Annotations

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