Alterations in arthropod and neuronal exosomes reduce virus transmission and replication in recipient cells

Abstract

Aim: Targeting the modes of pathogen shedding/transmission via exosomes or extracellular vesicles has been envisioned as the best approach to control vector-borne diseases. This study is focused on altering exosomes stability to affect the pathogen transmission from infected to naïve recipient cells.

Methods: In this study, neuronal or arthropod exosomes were treated at different temperatures or with different salts or pH conditions to analyze their ability and efficiency in the transmission of tick-borne Langat virus (LGTV) from infected to naïve recipient cells.

Results: Quantitative real-time PCR (qRT-PCR) and immunoblotting analyses revealed that treatment of neuronal or tick exosomes at warmer temperatures of 37 °C or 23 °C, respectively, or with sulfate salts such as Magnesium or Ammonium sulfates or with highly alkaline pH of 9 or 11.5, dramatically reduced transmission of LGTV via infectious exosomes (human or tick cells-derived) to human neuronal (SH-SY5Y) cells or skin keratinocytes (HaCaT cells), respectively.

Conclusion: Overall, this study suggests that exosome-mediated viral transmission of vector-borne pathogens to the vertebrate host or the viral dissemination and replication within or between the mammalian host can be reduced by altering the ability of exosomes with basic changes in temperatures, salts or pH conditions.

Keywords

Exosomes / neurons / skin cells / tick / Langat virus / transmission / treatments

References

[1] Sultana H.Arthropod exosomes as bubbles with message(s) to transmit vector-borne diseases.Curr Opin Insect Sci2020;40:39-47

[2] Zhou W,Sherman MB,Sultana H.Exosomes mediate Zika virus transmission through SMPD3 neutral Sphingomyelinase in cortical neurons.Emerg Microbes Infect2019;8:307-26 PMCID:PMC6455149

[3] Zhou W,Neupane B.Exosomes serve as novel modes of tick-borne flavivirus transmission from arthropod to human cells and facilitates dissemination of viral RNA and proteins to the vertebrate neuronal cells.PLoS Pathog2018;14:e1006764 PMCID:PMC5754134

[4] Vora A,Londono-Renteria B.Arthropod EVs mediate dengue virus transmission through interaction with a tetraspanin domain containing glycoprotein Tsp29Fb.Proc Natl Acad Sci USA2018;115:E6604-13 PMCID:PMC6048473

[5] Sampey GC,Zadeh MA,Hakami RM.Exosomes and their role in CNS viral infections.J Neurovirol2014;20:199-208 PMCID:PMC4378677

[6] Martin-Jaular L,Ferrer M,Del Portillo HA.Exosomes from Plasmodium yoelii-infected reticulocytes protect mice from lethal infections.PLoS One2011;6:e26588 PMCID:PMC3202549

[7] Bansal S,Lee J.Circulating exosomes induced by respiratory viral infections in lung transplant recipients activate cellular stress, innate immune pathways and epithelial to mesenchymal transition.Transpl Immunol2021;69:101480

[8] Rezaie J,Ahmadi M,Kashanchi F.The versatile role of exosomes in human retroviral infections: from immunopathogenesis to clinical application.Cell Biosci2021;11:19 PMCID:PMC7810184

[9] Wang Q,Zhen F,Meng F.Remedial applications of exosomes in cancer, infections and diabetes.Acta Pol Pharm2017;74:313-20

[10] Zhang W,Bao J,Liu H.Exosomes in pathogen infections: a bridge to deliver molecules and link functions.Front Immunol2018;9:90 PMCID:PMC5816030

[11] Jungbauer A.Exosomes enter vaccine development: strategies meeting global challenges of emerging infections.Biotechnol J2018;13:e1700749

[12] Coakley G,Buck AH.Exosomes and other extracellular vesicles: the new communicators in parasite infections.Trends Parasitol2015;31:477-89 PMCID:PMC4685040

[13] Landis JT,Pan Y.Evidence for multiple subpopulations of herpesvirus-latently infected cells.mBio2022;:e0347321 PMCID:PMC8725583

[14] Huang HJ,Mobley CM.Early humoral immune response to two doses of severe acute respiratory syndrome coronavirus 2 vaccine in a diverse group of solid organ transplant candidates and recipients.Clin Transplant2022;36:e14600

[15] Ouoba S,Nagashima S.Sequential dynamics of virological and serological changes in the serum of SARS-CoV-2 infected patients.J Med Virol2022;94:1734-7 PMCID:PMC9299762

[16] Gilbert PB,McDermott AB.Immune Assays Team§, Moderna, Inc. Team§., Coronavirus Vaccine Prevention Network (CoVPN)/Coronavirus Efficacy (COVE) Team§., United States Government (USG)/CoVPN Biostatistics Team§. Immune correlates analysis of the mRNA-1273 COVID-19 vaccine efficacy clinical trial.Science2022;375:43-50 PMCID:PMC9017870

[17] Hayashi T,Hutin YJ.The global hepatitis delta virus (HDV) epidemic: what gaps to address in order to mount a public health response?.Arch Public Health2021;79:180 PMCID:PMC8525025

[18] Earnest R,Matluk N.New England Variant Investigation TeamComparative transmissibility of SARS-CoV-2 variants Delta and Alpha in New England, USA.medRxiv2021;preprint PMCID:PMC8509091

[19] Thom RE,O’brien LM.Evaluation of the SARS-CoV-2 inactivation efficacy associated with buffers from three kits used on high-throughput RNA extraction platforms.Front Cell Infect Microbiol2021;11:716436

[20] Watson RJ,Fotheringham SA.Dose-dependent response to infection with ebola virus in the ferret model and evidence of viral evolution in the eye.J Virol2021;95:e0083321 PMCID:PMC8610581

[21] Johnston D,McCabe MS.Elucidation of the host bronchial lymph node miRNA transcriptome response to bovine respiratory syncytial virus.Front Genet2021;12:633125 PMCID:PMC8100685

[22] Johnston D,McCabe MS.Messenger RNA biomarkers of Bovine Respiratory Syncytial Virus infection in the whole blood of dairy calves.Sci Rep2021;11:9392 PMCID:PMC8087838

[23] Neelakanta G.Tick saliva and salivary glands: what do we know so far on their role in arthropod blood feeding and pathogen transmission.Front Cell Infect Microbiol2021;11:816547 PMCID:PMC8809362

[24] Zhou W,Wang JC.Discovery of exosomes from tick saliva and salivary glands reveals therapeutic roles for CXCL12 and IL-8 in wound healing at the tick-human skin interface.Front Cell Dev Biol2020;8:554 PMCID:PMC7378379

[25] Regmi P,Neelakanta G.Tick-borne flavivirus inhibits sphingomyelinase (IsSMase), a venomous spider ortholog to increase sphingomyelin lipid levels for its survival in Ixodes scapularis ticks.Front Cell Infect Microbiol2020;10:244 PMCID:PMC7325911

[26] Rego ROM,Anguita J.Counterattacking the tick bite: towards a rational design of anti-tick vaccines targeting pathogen transmission.Parasit Vectors2019;12:229 PMCID:PMC6518728

[27] Ahmed W,Neelakanta G.An experimental murine model to study acquisition dynamics of tick-borne langat virus in Ixodes scapularis.Front Microbiol2022;13:849313 PMCID:PMC9048798

[28] Maqbool M,Saqib M.Potential mechanisms of transmission of tick-borne viruses at the virus-tick interface.Front Microbiol2022;13:846884 PMCID:PMC9121816

[29] Santos RI,Reynolds ES.Salivary gland extract from the deer tick, Ixodes scapularis, facilitates neuroinvasion by Powassan virus in BALB/c mice.Sci Rep2021;11:20873 PMCID:PMC8536725

[30] Hart CE.Tick-virus interactions: current understanding and future perspectives.Parasite Immunol2021;43:e12815

[31] Hart CE,Kazimirova M.Tick-borne encephalitis virus infection alters the sialome of ixodes ricinus ticks during the earliest stages of feeding.Front Cell Infect Microbiol2020;10:41 PMCID:PMC7041427

[32] Boulanger N.Induced transient immune tolerance in ticks and vertebrate host: a keystone of tick-borne diseases?.Front Immunol2021;12:625993 PMCID:PMC7907174

[33] Wikel S.Immunobiology of tick-host-pathogen interactions.Parasite Immunol2021;43:e12818

[34] Wikel SK.Tick-host-pathogen systems immunobiology: an interactive trio.Front Biosci2018;23:265-83

[35] Wikel S.Ticks and tick-borne pathogens at the cutaneous interface: host defenses, tick countermeasures, and a suitable environment for pathogen establishment.Front Microbiol2013;4:337 PMCID:PMC3833115

[36] Chen G,Sohail M.Ixodes scapularis saliva mitigates inflammatory cytokine secretion during Anaplasma phagocytophilum stimulation of immune cells.Parasit Vectors2012;5:229 PMCID:PMC3503595

[37] Krause PJ,Tahan SR.Dermatologic changes induced by repeated Ixodes scapularis bites and implications for prevention of tick-borne infection.Vector Borne Zoonotic Dis2009;9:603-10 PMCID:PMC2883515

[38] Müller-Doblies UU,Boppana VD.Feeding by the tick, Ixodes scapularis, causes CD4(+) T cells responding to cognate antigen to develop the capacity to express IL-4.Parasite Immunol2007;29:485-99

[39] Pagel Van Zee J,Guerrero FD.Tick genomics: the Ixodes genome project and beyond.Int J Parasitol2007;37:1297-305

[40] Neelakanta G.Transmission-blocking vaccines: focus on anti-vector vaccines against tick-borne diseases.Arch Immunol Ther Exp2015;63:169-79 PMCID:PMC4429137

[41] Mans BJ.The basis of molecular diagnostics for piroplasmids: do the sequences lie?.Ticks Tick Borne Dis2022;13:101907

[42] Reck J,Dall'Agnol B.Transcriptomic analysis of salivary glands of ornithodoros brasiliensis Aragão, 1923, the agent of a neotropical tick-toxicosis syndrome in humans.Front Physiol2021;12:725635 PMCID:PMC8378177

[43] Pienaar R,de Castro MH,Mans BJ.De novo assembled salivary gland transcriptome and expression pattern analyses for Rhipicephalus evertsi evertsi Neuman, 1897 male and female ticks.Sci Rep2021;11:1642 PMCID:PMC7810686

[44] Mans BJ.Quantitative visions of reality at the tick-host interface: biochemistry, genomics, proteomics, and transcriptomics as measures of complete inventories of the tick sialoverse.Front Cell Infect Microbiol2020;10:574405 PMCID:PMC7517725

[45] Ribeiro JMC.TickSialoFam (TSFam): a database that helps to classify tick salivary proteins, a review on tick salivary protein function and evolution, with considerations on the tick sialome switching phenomenon.Front Cell Infect Microbiol2020;10:374 PMCID:PMC7396615

[46] Grabowski JM,Roumani AM.Changes in the proteome of langat-infected ixodes scapularis ISE6 cells: metabolic pathways associated with flavivirus infection.PLoS Negl Trop Dis2016;10:e0004180 PMCID:PMC4747643

[47] Grabowski JM,Kuhn RJ.RNAi reveals proteins for metabolism and protein processing associated with Langat virus infection in Ixodes scapularis (black-legged tick) ISE6 cells.Parasit Vectors2017;10:24 PMCID:PMC5237174

[48] Ahmed W,Sultana H.Tetraspanins as potential therapeutic candidates for targeting flaviviruses.Front Immunol2021;12:630571 PMCID:PMC8097176

[49] Rajendran KV,Sultana H.Sphingomyelinases in a journey to combat arthropod-borne pathogen transmission.FEBS Lett2021;595:1622-38

[50] Kurtti TJ,Herron MJ.Transgene expression and silencing in a tick cell line: a model system for functional tick genomics.Insect Biochem Mol Biol2008;38:963-8 PMCID:PMC2581827

[51] Oliver JD,Felsheim RF,Munderloh UG.An Ixodes scapularis cell line with a predominantly neuron-like phenotype.Exp Appl Acarol2015;66:427-42 PMCID:PMC4449809

[52] Chambers TJ.Pathogenesis of flavivirus encephalitis.Adv Virus Res2003;60:273-342 PMCID:PMC7202458

[53] Neal JW.Flaviviruses are neurotropic, but how do they invade the CNS?.J Infect2014;69:203-15

[54] Mukhopadhyay S,Rossmann MG.A structural perspective of the flavivirus life cycle.Nat Rev Microbiol2005;3:13-22

[55] Sips GJ,Smit JM.Neuroinvasive flavivirus infections.Rev Med Virol2012;22:69-87

[56] Alem F,Omole S.Exosomes originating from infection with the cytoplasmic single-stranded RNA virus Rift Valley fever virus (RVFV) protect recipient cells by inducing RIG-I mediated IFN-B response that leads to activation of autophagy.Cell Biosci2021;11:220 PMCID:PMC8710069

[57] Kim Y,Al Sharif S.Extracellular vesicles from infected cells are released prior to virion release.Cells2021;10:781 PMCID:PMC8066806

[58] Al Sharif S,Mensah GA.Extracellular vesicles in HTLV-1 communication: the story of an invisible messenger.Viruses2020;12:1422 PMCID:PMC7763366

[59] Pulliam L,Sun B.Using neuronal extracellular vesicles and machine learning to predict cognitive deficits in HIV.J Neurovirol2020;26:880-7 PMCID:PMC7718328

[60] Sun B,Pulliam L.Profile of neuronal exosomes in HIV cognitive impairment exposes sex differences.AIDS2019;33:1683-92

[61] Sun B,Abadjian L,Pulliam L.Blood neuron-derived exosomes as biomarkers of cognitive impairment in HIV.AIDS2017;31:F9-F17 PMCID:PMC5578870

[62] Pulliam L.Modulation of cellular function through immune-activated exosomes.DNA Cell Biol2015;34:459-63 PMCID:PMC4504252

[63] Gupta A.Exosomes as mediators of neuroinflammation.J Neuroinflammation2014;11:68 PMCID:PMC3994210

[64] Cheng Y,Han Q.Effect of pH, temperature and freezing-thawing on quantity changes and cellular uptake of exosomes.Protein Cell2019;10:295-9 PMCID:PMC6418301

[65] Liu X,Xing M,Li J.pH-mediated clustering of exosomes: breaking through the size limit of exosome analysis in conventional flow cytometry.Nano Lett2021;21:8817-23

[66] Li M,Yang S.Exosomes derived from bone marrow mesenchymal stem cells prevent acidic pH-induced damage in human nucleus pulposus cells.Med Sci Monit2020;26:e922928 PMCID:PMC7257871

[67] Du K,Tang X,Duan P.Effects of storage temperature and time on quality of plasma exosomes extracted by ExoQuick(TM) and Umibio kits.Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi2020;36:330-6

[68] Charoenviriyakul C,Nishikawa M.Preservation of exosomes at room temperature using lyophilization.Int J Pharm2018;553:1-7

[69] Yuan F,Wang Z.Preserving extracellular vesicles for biomedical applications: consideration of storage stability before and after isolation.Drug Deliv2021;28:1501-9 PMCID:PMC8281093

[70] Kumeda N,Akimoto Y,Tsujimoto M.Characterization of membrane integrity and morphological stability of human salivary exosomes.Biol Pharm Bull2017;40:1183-91

[71] Sokolova AS,Baranova DV.Quaternary ammonium salts based on (-)-borneol as effective inhibitors of influenza virus.Arch Virol2021;166:1965-76 PMCID:PMC8116641

[72] Kaushik N,Anang S.Zinc salts block hepatitis E virus replication by inhibiting the activity of viral RNA-dependent RNA polymerase.J Virol2017;91:e00754-17 PMCID:PMC5640865

[73] Macleod MK,Jin N.Influenza nucleoprotein delivered with aluminium salts protects mice from an influenza A virus that expresses an altered nucleoprotein sequence.PLoS One2013;8:e61775 PMCID:PMC3629017

[74] Wei Z,Palissa C,Schmidt MF.Antiviral activity of zinc salts against transmissible gastroenteritis virus in vitro.Vet Microbiol2012;160:468-72 PMCID:PMC7117232

[75] Bourne N,Montano R,Stanberry LR.Efficacy and toxicity of zinc salts as candidate topical microbicides against vaginal herpes simplex virus type 2 infection.Antimicrob Agents Chemother2005;49:1181-3 PMCID:PMC549274

[76] Suara RO.Effect of zinc salts on respiratory syncytial virus replication.Antimicrob Agents Chemother2004;48:783-90 PMCID:PMC353050

[77] Arens M.Zinc salts inactivate clinical isolates of herpes simplex virus in vitro.J Clin Microbiol2000;38:1758-62 PMCID:PMC86580

[78] Sergio W.Zinc salts that may be effective against the AIDS virus HIV.Med Hypotheses1988;26:251-3

[79] Wickerhauser M.The effects of salts on virus inactivation by lyophilization and dry heat.Vox Sang1987;53:188-9

[80] Lebeurier G,Hirth L.Effect of salts on tobacco mosaic virus reconstitution.FEBS Letters1973;35:54-8

[81] Ban JJ,Im W.Low pH increases the yield of exosome isolation.Biochem Biophys Res Commun2015;461:76-9