Fig. 3.
Extracellular vesicles from HIV-1-infected human tissues upregulate cytokines.
Extracellular vesicles (EVs) collected from donor-matched tissue cultures either infected with HIV-1, infected with HIV-1 and treated with ritonavir (RTV), treated only with RTV or uninfected (controls), and isolated using membrane affinity spin columns, were added to previously untreated (recipient) tissues and the release of cytokines was evaluated. EVs were collected from donor cultures on day 9 and added to the recipient cultures at day 0 and again on day 6 of culture. To exclude the possible effect of HIV-1 contamination in EVs collected from HIV-1-infected cultures, RTV was added to the tissue culture. Cytokine and p24 production were measured every 3 days and cumulative production was calculated. (a) HIV-1 replication in donor-matched tissues infected with HIV-1 or exposed to EVs. HIV-1 replication was assessed by p24gag released into the supernatant. Presented are mean cumulative releases (± SEM) over a 12 day period. (b--f) Cytokine release into the culture medium by tissues treated with EVs. Cytokine release is cumulative production over 12 days expressed in pg/ml. Presented are mean (±SEM) values (∗P < 0.05, n = 9).