10.1371/journal.pone.0032197.g004 Olivier Moralès Olivier Moralès Audrey Richard Audrey Richard Nathalie Martin Nathalie Martin Dhafer Mrizak Dhafer Mrizak Magalie Sénéchal Magalie Sénéchal Céline Miroux Céline Miroux Véronique Pancré Véronique Pancré Jean Rommelaere Jean Rommelaere Perrine Caillet-Fauquet Perrine Caillet-Fauquet Yvan de Launoit Yvan de Launoit Nadira Delhem Nadira Delhem Unlike PBMCs, CD4+ T cells do not undergo any major impairment upon H-1 PV infection. Public Library of Science 2013 cells impairment h-1 pv 2013-02-20 09:00:40 Figure https://plos.figshare.com/articles/figure/_Unlike_PBMCs_CD4_T_cells_do_not_undergo_any_major_impairment_upon_H_1_PV_infection_/352753 <p>CD4+ T cells were inoculated with increasing amounts of H-1 PV or mock-treated. MOI : multiplicity of infection, expressed as the number of plate-forming unit/cell; p.i : post-infection. <b>A</b>. <i>CD4+ T cell proliferation ability is not altered upon H-1 PV infection</i>. Cell proliferation was assessed by metabolic incorporation of tritiated thymidine into cellular DNA. Only 48 and 72 h p.i conditions are shown. Results are represented as means of triplicate wells with ± standard deviation bars and expressed in count per minute (cpm). <b>B</b>. <i>H-1 PV does not affect CD4+ T cell viability</i>. Cell viability was evaluated using a test based on a bioluminescent reaction measuring the amount of ATP present in living cells. Only 48 and 72 h p.i conditions are shown. Results are represented as means of triplicate wells with ± standard deviation bars and expressed in relative light unit (RLU). <b>C</b>. <i>Infection of CD4 T cells by H-1 PV is not related to any morphological changes</i>. For each condition, pictures of cells were taken in the course of infection but only those taken 48 h after H-1 PV inoculation are shown. Scale bar = 100 µm. <b>D</b>. <i>H-1 PV does not modify CD4+ T cell basal lysis</i>. H-1 PV cytotoxicity was assessed using a test based on a bioluminescent reaction measuring the leak of a cellular marker from cytoplasm to culture medium, which reveals the loss of plasma membrane integrity. Results are represented as means of triplicate wells with ± standard deviation bars and expressed in relative light unit (RLU). <b>E</b>. <i>CD4+ T cells transiently express a slight amount of NS1 protei</i>n. Equal amounts of total CD4+ T cell proteins were separated by 4–12% SDS-PAGE and analyzed by Western blot for the presence of NS1 protein. Erk2 was used as a loading control. <i>Representative data from 2 independent experiments</i>.</p>