How CD4+ T Cells Transcriptional Profile Is Affected by Culture Conditions: Towards the Design of Optimal In Vitro HIV Reactivation Assays

CD4+ T 细胞转录谱如何受培养条件影响:设计最佳体外 HIV 再激活试验

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作者:Giuseppe Rubens Pascucci, Elena Morrocchi, Chiara Pighi, Arianna Rotili, Alessia Neri, Chiara Medri, Giulio Olivieri, Marco Sanna, Gianmarco Rasi, Deborah Persaud, Ann Chahroudi, Mathias Lichterfeld, Eleni Nastouli, Caterina Cancrini, Donato Amodio, Paolo Rossi, Nicola Cotugno, Paolo Palma

Conclusions

These results suggest that for studies aiming at quantifying the magnitude of biological mechanisms under T cell activation, the autologous plasma could better approximate the in vivo environment. Conversely, if the study aims at defining qualitative aspects, then RPMI culture could provide more evident results.

Methods

Unstimulated or stimulated CD4+ T cells of three young adults with perinatal HIV-infection were isolated from PBMCs before or after culture in RPMI medium or autologous plasma. Transcriptomes were sequenced using Oxford Nanopore technologies.

Results

Transcriptional profiles revealed the activation of similar pathways upon stimulation in both media with a higher magnitude of TCR cascade activation in CD4+ lymphocytes cultured in RPMI. Conclusions: These results suggest that for studies aiming at quantifying the magnitude of biological mechanisms under T cell activation, the autologous plasma could better approximate the in vivo environment. Conversely, if the study aims at defining qualitative aspects, then RPMI culture could provide more evident results.

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