Cell-associated HIV-1 RNA was quantified in peripheral CD4+ T cells by ddPCR (One-Step RT-ddPCR Advanced Kit for Probes, BioRad) from samples taken before (0 h) and at the end of each RMD infusion (4 h), at 8 and 24 h (+1 day) after RMD1, and 72 h (+3 days) and 7 days after RMD1−2−3. CA HIV-1 RNA was quantified using two different primers/probe sets annealing to the 5′LTR and GAG conserved regions of HIV-1, to circumvent potential primer mismatch in individuals' viral sequence as previously described (37 (link)). HIV-1 transcription levels were normalized to the housekeeping gene TATA-binding protein (TBP) and shown as relative to levels before RMD1.
Mothe B., Rosás-Umbert M., Coll P., Manzardo C., Puertas M.C., Morón-López S., Llano A., Miranda C., Cedeño S., López M., Alarcón-Soto Y., Melis G.G., Langohr K., Barriocanal A.M., Toro J., Ruiz I., Rovira C., Carrillo A., Meulbroek M., Crook A., Wee E.G., Miró J.M., Clotet B., Valle M., Martinez-Picado J., Hanke T., Brander C, & Moltó J. (2020). HIVconsv Vaccines and Romidepsin in Early-Treated HIV-1-Infected Individuals: Safety, Immunogenicity and Effect on the Viral Reservoir (Study BCN02). Frontiers in Immunology, 11, 823.
Cd4 t cells Cell Hiv 1 Housekeeping gene Primer Tata binding protein Transcription
Corresponding Organization :
Other organizations :
Universitat de Vic - Universitat Central de Catalunya, IrsiCaixa, Red de Investigación en Sida, Hospital Universitari Germans Trias i Pujol, Universitat Autònoma de Barcelona, Hospital Clínic de Barcelona, Universitat de Barcelona, Consorci Institut D'Investigacions Biomediques August Pi I Sunyer, Universitat Politècnica de Catalunya, Institut d'Investigació en Ciències de la Salut Germans Trias i Pujol, Jenner Institute, University of Oxford, Hospital de Sant Pau, Institut Català de Ciències del Clima, Institució Catalana de Recerca i Estudis Avançats, Kumamoto University
Cell-associated HIV-1 RNA levels in peripheral CD4+ T cells
control variables
Housekeeping gene TATA-binding protein (TBP)
controls
Samples taken before (0 h) RMD infusion as a baseline control
Two different primers/probe sets annealing to the 5′LTR and GAG conserved regions of HIV-1 to circumvent potential primer mismatch in individuals' viral sequence
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