V. Bordoni, M. Bibas, I. Abbate, D. Viola, G. Rozera, C. Agrati, A

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Bone marrow CD34+ progenitor cells may harbour HIV-DNA even in successfully treated patients V. Bordoni, M. Bibas, I. Abbate, D. Viola, G. Rozera, C. Agrati, A. Rinaldi, A. Amendola, A. Ammassari, M.R. Capobianchi, F. Martini Clinical Microbiology and Infection Volume 21, Issue 3, Pages 290.e5-290.e8 (March 2015) DOI: 10.1016/j.cmi.2014.11.003 Copyright © 2014 European Society of Clinical Microbiology and Infectious Diseases Terms and Conditions

FIG. 1 Flow cytometry analysis of purification method used to analyse HIV-DNA in CD34+ hematopoietic progenitor cells (HPC). (a) Method used to isolate CD34+ HPC. CD34+ cells were first enriched from bone marrow mononuclear cells (BM-MNC) using indirect magnetic labelling of Lineage (Lin) positive cells. Highly pure CD34+Lin- populations were then obtained by a positive selection of CD34+ HPC using a direct magnetic labelling of lineage negative populations. (b) One representative flow cytometry analysis showing the percentage of CD34+ and CD4+ cells on Lineage-depleted populations of BM-MNC and (c) the corresponding purity of the CD34+Lin- cells obtained. Clinical Microbiology and Infection 2015 21, 290.e5-290.e8DOI: (10.1016/j.cmi.2014.11.003) Copyright © 2014 European Society of Clinical Microbiology and Infectious Diseases Terms and Conditions

FIG. 2 Analysis of HIV-DNA in untreated and antiretroviral therapy (ART)-treated HIV patients. (a) Clinical and experimental data from enrolled subjects. (b) HIV-DNA levels in bone marrow, peripheral blood, and CD34+Lin- cells from untreated and ART-treated patients (left, middle, and right panel). Bars show medians with range. (c) Linear regression analysis between HIV-DNA levels in CD34+Lin- BM-HPCs and plasma HIV-RNA levels; r = 0.6528, p = 0.0333. HIV-DNA levels were measured by PCR and then normalized to cell equivalents by using hTERT as reference. Clinical Microbiology and Infection 2015 21, 290.e5-290.e8DOI: (10.1016/j.cmi.2014.11.003) Copyright © 2014 European Society of Clinical Microbiology and Infectious Diseases Terms and Conditions