<i>Ex-vivo</i> HIV-1 infection of tonsil histocultures increases the soluble levels of CCL2MCP-1, suPAR, c-suPAR and the number of uPAR+ cells.

<p>Tonsil blocks were left either uninfected (Nil) or were infected with R5 or X4 HIV-1 strain. Every 3 days the culture conditioned supernatants were tested for the levels of viral replication (RT activity), lactate dehydrogenase (LDH), CCL2/MCP-1, suPAR, uPA and PAI-1. The results are reported as fold vs. Nil (absolute values are shown in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070606#pone.0070606.s001" target="_blank">Figure S1</a>). Panels A (n = 31), B (n = 31) and C (n = 28) show fold of HIV-1 replication, LDH and CCL2/MCP-1 levels vs. Nil, respectively, as measured in the culture supernatants The panels below show the IHC detection of HIV p24_Gag measured 6 days post-infection and hematoxylin-eosin staining at day 9 after infection. Panel D (n = 28) shows fold of suPAR levels vs. Nil measured in the culture supernatants. Panel E (n = 7) shows the IHC score in terms of number of uPAR⁺ cells in Nil and <i>ex-vivo</i> infected tonsil histocultures at the indicated time points after infection (Day 0 is the time of surgical removal); the right panels show the IHC of uPAR+ cells (brown) 9 days post-infection. Panel F shows western blot analysis evaluating for the presence of both uPAR and c-uPAR and of HIV-1 p24 _Gag release in the culture supernatants of tonsils left uninfected (Nil) or infected <i>in vitro</i> for the indicated time points after infection. Panels G (n = 21) and panel H (n = 21) show the fold of uPA and PAI-1 levels vs. Nil measured in the culture supernatants. The vertical and horizontal bars represent the mean and SEM. Black dots and triangles in panels A-E and G-H represent data from tonsils infected <i>ex-vivo</i> with R5 and X4 strain, respectively; white circles in panel E represent data from uninfected ex-vivo tonsils. Statistical significance is indicated by asterisks (*** = <0.001); 2-tailed paired t test (panel A), 2-tailed Wilcoxon signed-rank test (panels C and D). Pictures are at 40×magnification.