Volume: 45 Issue: 3
Year: 2025, Page: 200-208, Doi: https://doi.org/10.51248/v45i3.114
Received: May 19, 2025 Accepted: Aug. 30, 2025 Published: July 9, 2025
The research evaluates two in-vitro approaches for HIV-1 culture from stored samples, viz. infection of donor PBMC with patient plasma by spinoculation and co-culture of patient PBMC with donor PBMC. The evaluation of virus production depended on detection of p24 antigen in culture supernatant and infection of TZM-bl reporter cells. Most of the samples infected using spinoculation method showed p24 absorbance levels exceeding the threshold value on day 7. The levels peaked by days 9/10 and decreased by day 13. The TZM-bl assay gave similar results. The PBMC co-culture methodology resulted in a steady increase in p24 antigen levels starting from day 5 and continued to increase gradually the levels of peak achieved by day 9 and decreased by day 13 over time. The results of our study indicate that spinoculation enables virus production from stored plasma, yet co-culture of PBMCs appears to be a more robust approach for virus amplification.
Keywords: Spinoculation, PBMC co-culture, HIV-1, TZM bl infectivity assay, p24 antigen detection, ELISA
Tammanna Bhajantri, S. Pushkala, Luke Elizabeth Hanna, Lucia Precilla, Malathi M. Comparison of Spinoculation and Coculture Techniques for Facilitating HIV-1 Infection of Healthy PBMCs Using Patient-Derived PBMCs and Plasma. Biomedicine: 2025, 45(3): 200-208