Work

From phlebotomy to peripheral blood mononuclear cell isolation; effect of time and early-life HIV/ART exposure on cell yield and viability in paediatric samples in a high HIV prevalence setting

Year: 2025

Type: article

Abstract: BACKGROUND: Viral infection/exposure and time from phlebotomy to peripheral blood mononuclear cell (PBMC) isolation (TPP) may affect PBMC yield and viability. We investigated the impact of TPP and early-life HIV/ART exposure on PBMC, including NK cells yield and viability. METHODS: Blood in EDTA tubes with varying TPP was used to isolate PBMCs via density-gradient centrifugation, counted under a microscope, and assessed for viability with Trypan-Blue. Antibody staining (CD14, CD3, and CD19) excluded monocytes and T/B lymphocytes, while CD16 and CD56 staining with a LIVE/DEAD marker identified viable NK cells. Comparisons were made among HIV-exposed uninfected (HEU) children (long-term [HEULT] and medium-term [HEUMT] ART exposure), HIV-exposed infected (HEI), and HIV-unexposed uninfected (HUU) children. RESULTS: cells/ml) (p = 0.010) for all children. Overall, TPP negatively correlated with yields/ml and viability (r = -0.35, p < 0.001; r = -0.47, p < 0.001 respectively). Median frequency of viable CD56 + CD16+/- NK cells was 8.2 %, unaffected by neither TPP nor HIV/ART exposure. Total lymphocytes were lower in males [18.6 % (IQR:11.3-22.2)] than in females [23.8 % (IQR:14.3-29.9)] (p = 0.018). CONCLUSION: HIV infection, not exposure, may increase PBMC yields. TPP under 3 h is ideal for optimal yields.

Source: Journal of Immunological Methods

DOI: 10.1016/j.jim.2025.113906

Authors: Hope Mataramvura, Patience Ncube, Kerina Duri

Institution: University of Zimbabwe

Language: English

Topic: HIV Research and Treatment

Subfield: Virology

Field: Immunology and Microbiology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: hybrid

Estimated APC paid: $2,970