Cytokine profiling of exosomes derived from the plasma of HIV-infected alcohol drinkers and cigarette smokers

Abstract

Cytokines and chemokines circulate in plasma and may be transferred to distant sites, via exosomes. HIV infection is associated with dysregulation of cytokines and chemokines, which subsequently contribute to the pathogenesis of HIV. Alcohol and tobacco exposure, which are prevalent in HIV-infected individuals, may induce changes in the expression of cytokines and chemokines. Therefore, our aim in this study was to quantify plasma exosomal cytokines and chemokines that we expect to exacerbate toxicity or disease progression in HIV-positive drug abusers. We measured the levels of cytokines and chemokines in the plasma and plasma exosomes of 39 patients comprising six groups: HIV-negative and HIV-positive non drug abusers, HIV-negative and HIV-positive alcohol users, and HIV-negative and HIV positive tobacco smokers. We measured six cytokines (TNF-α, IL-1β, IL-8, IL-6, IL-1ra, IL-10) and two chemokines (MCP-1 and RANTES). All were present in exosomes of healthy subjects, but their levels varied between different study groups. HIV-positive alcohol drinkers had higher levels of plasma IL-8 compared to those of HIV-positive non-drinkers. The IL-1ra level was significantly higher in exosomes of non-HIV-infected alcohol drinkers compared to those of HIV-positive alcohol drinkers. Interestingly, the IL-10 level was higher in exosomes compared with their respective plasma levels in all study groups except HIV-positive non-alcohol drinkers. IL-10 was completely packaged in exosomes of HIV-positive smokers. HIV-positive smokers had significantly higher levels of plasma IL-8 compared with HIV-positive non-smokers and significantly higher exosomal IL-6 levels compared with HIV-negative subjects. HIV-positive smokers had significantly increased plasma levels of IL-1ra compared to HIV-positive non-smokers. The MCP-1 levels in the plasma of HIV-positive smokers was significantly higher than in either HIV-positive non-drug abusers or HIV-negative smokers. Overall, the findings suggest that plasma cytokines and chemokines are packaged in exosomes at varying degrees in different study groups. Exosomal cytokines and chemokines are likely to have a significant biological role at distant sites including cells in the brain.

Extracellular Vesicles in Smoking-Mediated HIV Pathogenesis and their Potential Role in Biomarker Discovery and Therapeutic Interventions

“…We have also looked at the differential cytokine packaging within the same population [37]. We identified two cytokine/chemokines, IL-6 and MCP-1, which were significantly packaged within plasma EVs from HIV-infected smokers [37]. We further confirmed this finding by measuring their concentrations in the EVs derived from macrophages exposed to CSC and HIV, where IL-6 showed a similar trend (manuscript under review).…”

“…We also observed an alteration in the levels of CD9, CETP, and vitronectin (VTN) in plasma EVs from the HIV positive population; however, further studies are required to reach to any conclusion. We have also looked at the differential cytokine packaging within the same population [37]. We identified two cytokine/chemokines, IL-6 and MCP-1, which were significantly packaged within plasma EVs from HIV-infected smokers [37].…”

“…EVs play an important role in HIV pathogenesis -either in improving or deteriorating the existing condition; however, the exact role of EVs in HIV pathogenesis is poorly understood [35,36]. Currently, only a handful of studies have investigated the role of EVs in smoking-mediated toxicity in the setting of HIV [37][38][39].…”

“…Conversely, HIV-infected macrophage-derived EVs lose this defense capacity, as evidenced by a higher viral load and increased cellular toxicity [39]. Proteomic and cytokine analyses of plasma EVs obtained from HIV-positive and negative smokers demonstrated a differential packaging of proteins in the EVs [37,49]. In addition, macrophage-derived EVs can readily cross the blood brain barrier, suggesting the potential role of EVs in either disseminating or alleviating HIV and HAND pathogenesis [30].…”

Extracellular Vesicles from Human Papilloma Virus-Infected Cervical Cancer Cells Enhance HIV-1 Replication in Differentiated U1 Cell Line

“…HPV16 E6 and E7 oncoproteins are known to cause chronic oxidative stress in HPV-infected and uninfected cells via NOX2 activation [14]. In addition, CCS-derived EVs may also contain various cytokines [56] and miRNAs that may trigger HIV-1 replication in differentiated U1 cell line. Cytokines, such as TNF-α, trigger HIV-1 gene transcription through activation of NF-κB; IL-6 increases the expression of viral proteins and RT; IL-6, together with TNF-α, have a synergistic effect on HIV-1 replication [57].…”