Fig. 1: Development of the MDM-QVOA.
From: Monocyte-derived macrophages contain persistent latent HIV reservoirs
a, To determine the appropriate expander cell line, one viremic vPWH was activated with PMA in the context of MT-4, Molt-4-CCR5, CEMx174 and media only, dotted line indicates limit of detection (LOD). b, To determine the best activation condition, 7 participants (n = 7, 4 vPWH and 3 virally suppressed vsPWH) were activated with PMA, IL-4, TNF𝛼 and media with and without MT-4 expander cells; mean ± s.d. c, To determine whether macrophages derived from negatively selected monocytes could be reactivated similarly to macrophages derived from whole PBMCs, we compared 3 participants (n = 3, 1 vPWH and 2 vsPWH) activated with PMA and co-cultured with MT-4. d–f, To determine the appropriate assay to detect T cell contamination, in the well or via phagocytosis, we assessed CD3ε and TCRβ RNA expression in CD4 T cells isolated from healthy donors (HD). d, The CD4 T cells from 3 HD were serially diluted, lysed and RNA extracted to measure TCRβ and CDε expression; mean ± s.d. TCRβ (e) and CDε (f) showed similar variability across replicates, except at the low end of the assay. g, CD4 T cells were isolated from 8 HD; 1 × 106 CD4s per donor were lysed and assessed for CD3ε and TCRβ expression to determine the copies of each per cell; bar indicates median value. h, Healthy MDMs were co-cultured with HIV+ CD4 T cells from two donors (CP11 and 21) with and without PMA activation to determine whether HIV+ CD4 T cells were able to transfer viral nucleic acids to MDM; n = 2. i, A schematic of the final MDM-QVOA experimental design.
