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Updated: Jun 9, 2026
Chronic, Acute, and Reactivated HIV Infection in Humanized Immunodeficient Mouse Models
Published on: December 3, 2019
Pooled peripheral blood mononuclear cells provide an optimized cellular substrate for human immunodeficiency virus
Christopher Lai-Hipp1, Tiffany Goldberg, Edward Scott
1Department of Laboratory Medicine, University of California, San Francisco, California, USA.
Background:
Cellular substrate (CS) composed of phytohemagglutinin (PHA)-stimulated PBMNCs is required for isolating primary human immunodeficiency virus Type 1 from plasma (PHIV) during early acute infection. The transmitted founder PHIV is neutralization-sensitive and uses chemokine-receptor 5 to infect CD4-positive PBMNCs. Therefore, PHIV cocultured with optimized CS would enable efficient virologic diagnosis, vaccine development, and tests for broadly neutralizing antibodies (bnAb).
Study Design And Methods:
Fifteen leukapheresis donations were used to isolate CD4-enriched PBMNCs. Individual or three or four donors' pooled cells were stimulated with PHA, and both CSs were evaluated in parallel cocultures using five different PHIV isolates from nucleic acid testing-positive anti-HIV-negative donations. Feasibility of coculturing PHIV with pooled PBMNC-CS after dimethyl sulfoxide cryopreservation was investigated.
Results:
The cell-free supernatants from individual CSs contained mean HIV-p24 antigen varying between 20.35 and 85.54 ng/mL, while that from pooled CSs ranged between 57.54 and 88.10 ng/mL. Thus, PBMNCs pooled from multiple donors provide an optimal CS for coculturing PHIV. Cryopreserved PBMNC-CS for PHIV cocultures is promising for systematic biosynthesis of PHIV.
Conclusions:
Pooled CSs after cryopreservation were functional for PHIV replication and enable on-demand diagnostic cocultures, preparing panels of PHIV stocks for bnAb testing and developing an envelope-subunit vaccine candidate analogous to hepatitis B surface antigen, proven successful in preventing hepatitis B virus infection.
Insights
Optimized cellular substrate (CS) from pooled donor cells enhances primary HIV-1 isolation and replication. Cryopreserved CS enables on-demand diagnostics and vaccine development for HIV.
Area of Science:
- Virology
- Immunology
- Cell Biology
Background:
- Cellular substrate (CS) from phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMNCs) is crucial for isolating primary human immunodeficiency virus Type 1 (PHIV) during early acute infection.
- Transmitted founder PHIV utilizes chemokine receptor 5 (CCR5) to infect CD4-positive PBMNCs and is neutralization-sensitive.
Purpose of the Study:
- To optimize cellular substrate (CS) for efficient isolation and replication of primary HIV Type 1 (PHIV).
- To evaluate the potential of cryopreserved CS for diagnostic and therapeutic applications, including vaccine development and broadly neutralizing antibody (bnAb) testing.
Main Methods:
- Isolated CD4-enriched PBMNCs from fifteen leukapheresis donations.
- Stimulated individual or pooled PBMNCs with PHA to create CS.
- Evaluated CS in parallel cocultures with five different PHIV isolates.
- Investigated feasibility of cryopreservation for pooled PBMNC-CS.
Main Results:
- Pooled PBMNC-CS yielded higher mean HIV-p24 antigen levels (57.54–88.10 ng/mL) compared to individual CS (20.35–85.54 ng/mL), indicating optimal PHIV replication.
- Cryopreserved PBMNC-CS demonstrated feasibility for PHIV cocultures, suggesting potential for systematic PHIV biosynthesis.
Conclusions:
- Pooled cellular substrate (CS) from cryopreserved peripheral blood mononuclear cells (PBMNCs) is functional for PHIV replication.
- This optimized CS enables on-demand diagnostic cocultures, preparation of PHIV stocks for bnAb testing, and development of novel vaccine candidates.
