Differential packaging of inflammatory cytokines/ chemokines and oxidative stress modulators in U937 and U1 macrophages-derived extracellular vesicles upon exposure to tobacco constituents

Abstract

Smoking, which is highly prevalent in HIV-infected populations, has been shown to exacerbate HIV replication, in part via the cytochrome P450 (CYP)-induced oxidative stress pathway. Recently, we have shown that extracellular vesicles (EVs), derived from tobacco-and/ or HIV-exposed macrophages, alter HIV replication in macrophages by cell-cell interactions. We hypothesize that cigarette smoke condensate (CSC) and/or HIV-exposed macrophagederived EVs carry relatively high levels of pro-oxidant and pro-inflammatory cargos and/or low levels of antioxidant and anti-inflammatory cargos, which are key mediators for HIV pathogenesis. Therefore, in this study, we investigated differential packaging of pro-and anti-inflammatory cytokines/chemokines and pro-and anti-oxidant contents in EVs after CSC exposure to myeloid cells (uninfected U937 and HIV-infected U1 cells). Our results showed that relatively long to short exposures with CSC increased the expression of cytokines in EVs isolated from HIV-infected U1 macrophages. Importantly, pro-inflammatory cytokines, especially IL-6, were highly packaged in EVs isolated from HIV-infected U1 macrophages upon both long and short-term CSC exposures. In general, anti-inflammatory cytokines, particularly IL-10, had a lower packaging in EVs, while packaging of chemokines was mostly increased in EVs upon CSC exposure in both HIV-infected U1 and uninfected U937 macrophages. Moreover, we observed higher expression of CYPs (1A1 and 1B1) and lower expression of antioxidant enzymes (SOD-1 and catalase) in EVs from HIV-infected U1 macrophages than in uninfected U937 macrophages. Together, they are expected to increase oxidative stress factors in EVs derived from HIV-infected U1 cells. Taken together, our results suggest packaging of increased level of oxidative stress and inflammatory elements in the EVs upon exposure to tobacco constituents and/or HIV to myeloid cells, which would ultimately enhance HIV replication in macrophages via cell-cell interactions.

Extracellular Vesicles in Viral Replication and Pathogenesis and Their Potential Role in Therapeutic Intervention

“…Similar to the previous study, exposure to cigarette smoke condensate (CSC) increased the packaging of cytokines, especially IL-6 and CYPs (1A1 and 1B1) in EVs isolated from HIV-infected U1 macrophages [116]. Conversely, EV packaging of AOEs (SOD-1 and catalase) decreased in HIV-infected U1 macrophages more than in uninfected U937 macrophages [116]. Recently, our group showed that the astrocytic and neuronal-specific markers (GFAP and L1CAM) can be packaged in EVs and circulate in plasma, which is further elevated in the presence of HIV infection, alcohol, and/or tobacco [121].…”

“…However, exosomes derived from HIV-infected cells lost their protective capacity that could be due to the selective packaging of cytochrome P450 (CYPs) and antioxidant enzyme (AOE) mRNAs in exosomes [21]. Similar to the previous study, exposure to cigarette smoke condensate (CSC) increased the packaging of cytokines, especially IL-6 and CYPs (1A1 and 1B1) in EVs isolated from HIV-infected U1 macrophages [116]. Conversely, EV packaging of AOEs (SOD-1 and catalase) decreased in HIV-infected U1 macrophages more than in uninfected U937 macrophages [116].…”

“…The authors suggest that this occurs because ARV or interferon prevents the release of viral particles from cells, which then allows for viral RNA to be packaged into EVs due to the increased presence of viral RNA in the cell. In addition to viral RNA, a variety of molecules, e.g., viral & host proteins, cellular markers, miRNA, inflammatory molecules such as oxidative stress markers, chemokines and cytokines can also be packaged into EVs [20,[115][116][117]119,121]. A study showed that the viral envelop (Env) protein can be packaged into EVs from infected cells [147].…”

Extracellular Vesicles in Viral Infections: Two Sides of the Same Coin?

“…During infection, EVs can amplify inflammation and deflagrate antiviral responses (Urbanelli et al, 2019) and can also mediate communication between immune cells and other cell types (Isola and Chen, 2017). The involvement of EVs in viral infection and/or host interactions in disease has already been described for several viruses, such as rabies (Wang et al, 2019b), coronaviruses (Maeda et al, 1999;Kuate et al, 2007;Börger et al, 2020;Deffune et al, 2020;Hassanpour et al, 2020;Inal, 2020a;Inal, 2020b;Kumar et al, 2020;O'Driscoll, 2020;Tsuchiya et al, 2020;Urciuoli and Peruzzi, 2020), HCV (Bartosch et al, 2003;Timpe et al, 2008;Dreux et al, 2012;Bukong et al, 2014), HBV (Jia et al, 2017;, HIV (Princen et al, 2004;Khatua et al, 2009;Xu et al, 2009;Lenassi et al, 2010;Bernard et al, 2014;Raymond et al, 2016;Sampey et al, 2016;Kodidela et al, 2018;Haque et al, 2020;Ranjit et al, 2020), HPV (Honegger et al, 2015;Guenat et al, 2017;Sadri Nahand et al, 2019;Chiantore et al, 2020), HSV (Temme et al, 2010;Han et al, 2016;Deschamps and Kalamvoki, 2018) dengue (Martins et al, 2018;…”

Anti-HIV Activity of Cucurbitacin-D against Cigarette Smoke Condensate-Induced HIV Replication in the U1 Macrophages

“…U1 cells, which are U937 cells chronically infected with the human immunodeficiency virus type 1 (HIV-1), were procured from the NIH AIDS Reagent Program (Germantown, MD, USA). The U1 cell lines have been widely used by our group and many other research groups to study the role of drug abuse, including tobacco smoking in HIV replication [10,11,[22][23][24][25]. Furthermore, the results obtained using these cells have been successfully replicated in HIV-infected primary macrophages [10,11,24].…”