Milk exosomes elicit a potent anti-viral activity against dengue virus

Abstract

Background Exosomes are nano-sized vesicles secreted by various cells into the intra and extracellular space and hence is an integral part of biological fluids including milk. In the last few decades, many research groups have proved the potential of milk exosomes as a sustainable, economical and non-immunogenic drug delivery and therapeutic agent against different pathological conditions. However, its anti-viral properties still remain to be unearthed. Methods Here, we have been able to isolate, purify and characterize the milk derived exosomes from Cow (CME) and Goat (GME) and further studied its antiviral properties against Dengue virus (DENV), Newcastle Disease Virus strain Komarov (NDV-K) and Human Immunodeficiency Virus (HIV-1) using an in-vitro infection system. Results TEM, NTA and DLS analysis validated the appropriate size of the isolated cow and goat milk exosomes (30–150 nm). Real-time PCR and immunoblotting results confirmed the presence of several milk exosomal miRNAs and protein markers. Our findings suggest that GME significantly decreased the infectivity of DENV. In addition, we confirmed that GME significantly reduces DENV replication and reduced the secretion of mature virions. Furthermore, heat inactivation of GME did not show any inhibition on DENV infection, replication, and secretion of mature virions. RNase treatment of GME abrogates the anti-viral properties indicating direct role of exosomes in DENV inhibition. In addition GME inhibited the infectivity of NDV-K, but not HIV-1, suggesting that the GME mediated antiviral activity might be virus specific. Conclusion This study demonstrates the anti-viral properties of milk exosomes and opens new avenues for the development of exosome-based therapies to treat viral diseases. Graphical Abstract

Dual-labeled nanoparticles based on small extracellular vesicles for tumor detection

“…These milk-derived vesicles were selected not only on the basis of their structural characteristics, such as nanometric size, robustness, and lipid bilayer morphology, but also on their proven role in tumor pathology [32][33][34][35][36] or inflammatory processes, as shown in previous studies using goat milk sEVs as optical probes in a model of peritonitis [37] or in a recent study where their anti-viral activity was probed [38]. Moreover, milk EVs from other sources have been proposed as an excellent platform for drug delivery in cancer therapy [35], further supporting our application of these vesicles in the detection of tumor foci.…”

Exosomes in Mastitis—Research Status, Opportunities, and Challenges

“…For example, studies have confirmed that milk-derived exosomes can be absorbed by a variety of cells [ 89 , 119 ] and can be used as delivery vehicles for short interfering RNA (siRNA), miRNA, and drugs [ 120 , 121 ]. The antiviral activity [ 122 ], anti-inflammatory activity [ 123 , 124 , 125 ], and the anticancer activity [ 126 , 127 ] of milk-derived exosomes have also been characterized through experiments. Several groups have compared the functional activity of bovine, buffalo, goat, and yak milk-derived exosomes [ 128 , 129 , 130 , 131 ].…”

Extracellular Vesicles from Animal Milk: Great Potentialities and Critical Issues

“…Surprisingly, only a few studies have evaluated EVs from goat milk despite its extensive use in human nutrition and dairy products and the potential that emerged for these vesicles in terms of molecular cargo [ 101 , 112 ]. Indeed, in addition to the transcriptomic and metabolic characterisation of the cargo, to the authors’ knowledge, only two studies have evaluated the functionality of these mEVs, in particular their anti-inflammatory and immunomodulatory properties in an in vitro model of intestinal inflammation [ 185 ] and their antiviral properties against the dengue virus (DENV), Newcastle disease virus–Komarov strain (NDV-K), and human immunodeficiency virus (HIV-1) using an in vitro infection system [ 186 ]. Goat mEVs were able to significantly decrease the replication of DENV and infectivity of DENV and NDV-K but not HIV-1, which suggested a virus-specific activity [ 186 ].…”

“…Indeed, in addition to the transcriptomic and metabolic characterisation of the cargo, to the authors’ knowledge, only two studies have evaluated the functionality of these mEVs, in particular their anti-inflammatory and immunomodulatory properties in an in vitro model of intestinal inflammation [ 185 ] and their antiviral properties against the dengue virus (DENV), Newcastle disease virus–Komarov strain (NDV-K), and human immunodeficiency virus (HIV-1) using an in vitro infection system [ 186 ]. Goat mEVs were able to significantly decrease the replication of DENV and infectivity of DENV and NDV-K but not HIV-1, which suggested a virus-specific activity [ 186 ]. Moreover, goat mEVs seemed to have protective effects on enterocytes after LPS stimulation by reducing inflammation and inducing the expression of genes to restore the mucosal barrier homeostasis [ 185 ].…”