{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:29:53Z","timestamp":1760239793805,"version":"build-2065373602"},"reference-count":41,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2020,12,31]],"date-time":"2020-12-31T00:00:00Z","timestamp":1609372800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Military Infectious Diseases Research Program","award":["WJ0134_19"],"award-info":[{"award-number":["WJ0134_19"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Exosomes are small extracellular vesicles that carry proteins, lipids, and nucleic acids. They are circulated in many body fluids and play an important role in intercellular communications. MicroRNAs (miRNAs), as major components of exosomes, are often regulated in many diseases including bacterial and viral infections. Functionally, exosome-carried miRNAs interact with various immune cells and affect their behavior. Little is known whether exosomal miRNAs are regulated during scrub typhus, a potentially lethal infection caused by intracellular bacteria, Orientiatsutsugamushi. In the present study, we utilized a scrub typhus mouse model and collected serum at various time points post infection. A custom quantitative PCR array covering 92 murine miRNAs was used to profile serum exosomal miRNAs. A total of 12 miRNAs were found to be significantly up- or down-regulated at least at one time point post infection when compared to uninfected animals. Further analysis identified multiple miRNAs in the let-7 family that were consistently down-regulated at early and late phase of infection. Functionally, serum exosomes isolated from infected mice displayed strong proinflammatory effect when incubated with bone marrow-derived macrophages. Our data revealed dynamic regulations of serum exosomal miRNA during scrub typhus infection, which could significantly influence host immune responses and disease outcome.<\/jats:p>","DOI":"10.3390\/microorganisms9010080","type":"journal-article","created":{"date-parts":[[2020,12,31]],"date-time":"2020-12-31T10:10:37Z","timestamp":1609409437000},"page":"80","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Regulation of Serum Exosomal MicroRNAs in Mice Infected with Orientia tsutsugamushi"],"prefix":"10.3390","volume":"9","author":[{"given":"Le","family":"Jiang","sequence":"first","affiliation":[{"name":"Viral and Rickettsial Diseases Department, Infectious Diseases Directorate, Naval Medical Research Center, Silver Spring, MD 20910, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tatyana","family":"Belinskaya","sequence":"additional","affiliation":[{"name":"Viral and Rickettsial Diseases Department, Infectious Diseases Directorate, Naval Medical Research Center, Silver Spring, MD 20910, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhiwen","family":"Zhang","sequence":"additional","affiliation":[{"name":"Viral and Rickettsial Diseases Department, Infectious Diseases Directorate, Naval Medical Research Center, Silver Spring, MD 20910, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Teik-Chye","family":"Chan","sequence":"additional","affiliation":[{"name":"Viral and Rickettsial Diseases Department, Infectious Diseases Directorate, Naval Medical Research Center, Silver Spring, MD 20910, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei-Mei","family":"Ching","sequence":"additional","affiliation":[{"name":"Viral and Rickettsial Diseases Department, Infectious Diseases Directorate, Naval Medical Research Center, Silver Spring, MD 20910, USA"},{"name":"Department of Preventive Medicine and Biostatistics, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chien-Chung","family":"Chao","sequence":"additional","affiliation":[{"name":"Viral and Rickettsial Diseases Department, Infectious Diseases Directorate, Naval Medical Research Center, Silver Spring, MD 20910, USA"},{"name":"Department of Preventive Medicine and Biostatistics, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,12,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1097\/00001432-200310000-00009","article-title":"Scrub typhus and tropical rickettsioses","volume":"16","author":"Watt","year":"2003","journal-title":"Curr. Opin. Infect. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Jiang, J., and Richards, A.L. (2018). Scrub Typhus: No Longer Restricted to the Tsutsugamushi Triangle. Trop. Med. Infect. Dis., 3.","DOI":"10.3390\/tropicalmed3010011"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"301","DOI":"10.4269\/ajtmh.13-0064","article-title":"Unresolved problems related to scrub typhus: A seriously neglected life-threatening disease","volume":"89","author":"Paris","year":"2013","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Paris, D.H., Phetsouvanh, R., Tanganuchitcharnchai, A., Jones, M., Jenjaroen, K., Vongsouvath, M., Ferguson, D.P., Blacksell, S.D., Newton, P.N., and Day, N.P. (2012). Orientia tsutsugamushi in human scrub typhus eschars shows tropism for dendritic cells and monocytes rather than endothelium. PLoS Negl. Trop. Dis., 6.","DOI":"10.1371\/journal.pntd.0001466"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/j.micpath.2005.08.004","article-title":"Survival of two Orientia tsutsugamushi bacterial strains that infect mouse macrophages with varying degrees of virulence","volume":"39","author":"Fukuhara","year":"2005","journal-title":"Microb. Pathog."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Choi, J.H., Cheong, T.C., Ha, N.Y., Ko, Y., Cho, C.H., Jeon, J.H., So, I., Kim, I.K., Choi, M.S., and Kim, I.S. (2013). Orientia tsutsugamushi subverts dendritic cell functions by escaping from autophagy and impairing their migration. PLoS Negl. Trop. Dis., 7.","DOI":"10.1371\/journal.pntd.0001981"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Hauptmann, M., Kolbaum, J., Lilla, S., Wozniak, D., Gharaibeh, M., Fleischer, B., and Keller, C.A. (2016). Protective and Pathogenic Roles of CD8+ T Lymphocytes in Murine Orientia tsutsugamushi Infection. PLoS Negl. Trop. Dis., 10.","DOI":"10.1371\/journal.pntd.0004991"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1007\/s00018-017-2595-9","article-title":"Current knowledge on exosome biogenesis and release","volume":"75","author":"Hessvik","year":"2018","journal-title":"Cell Mol. Life Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1016\/j.immuni.2014.08.008","article-title":"MicroRNA-Containing T-Regulatory-Cell-Derived Exosomes Suppress Pathogenic T Helper 1 Cells","volume":"41","author":"Okoye","year":"2014","journal-title":"Immunity"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"17095","DOI":"10.1038\/s41598-017-16609-6","article-title":"Exosomes from antigen-pulsed dendritic cells induce stronger antigen-specific immune responses than microvesicles in vivo","volume":"7","author":"Wahlund","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"90","DOI":"10.3389\/fimmu.2018.00090","article-title":"Exosomes in Pathogen Infections: A Bridge to Deliver Molecules and Link Functions","volume":"9","author":"Zhang","year":"2018","journal-title":"Front. Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"936","DOI":"10.4269\/ajtmh.2005.73.936","article-title":"Protection against scrub typhus by a plasmid vaccine encoding the 56-KD outer membrane protein antigen gene","volume":"73","author":"Ni","year":"2005","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4550","DOI":"10.1016\/S0264-410X(03)00505-X","article-title":"Development of a rapid method for determining the infectious dose (ID)50 of Orientia tsutsugamushi in a scrub typhus mouse model for the evaluation of vaccine candidates","volume":"21","author":"Chan","year":"2003","journal-title":"Vaccine"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"351","DOI":"10.4269\/ajtmh.2004.70.351","article-title":"Development of a quantitative real-time polymerase chain reaction assay specific for Orientia tsutsugamushi","volume":"70","author":"Jiang","year":"2004","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Chao, C.C., Belinskaya, T., Zhang, Z., Jiang, L., and Ching, W.M. (2019). Assessment of a Sensitive qPCR Assay Targeting a Multiple-Copy Gene to Detect Orientia tsutsugamushi DNA. Trop. Med. Infect. Dis., 4.","DOI":"10.3390\/tropicalmed4030113"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"14888","DOI":"10.1073\/pnas.1408301111","article-title":"Quantitative and stoichiometric analysis of the microRNA content of exosomes","volume":"111","author":"Chevillet","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"42712","DOI":"10.18632\/oncotarget.17448","article-title":"Characterization of mouse serum exosomal small RNA content: The origins and their roles in modulating inflammatory response","volume":"8","author":"Zhou","year":"2017","journal-title":"Oncotarget"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"R64","DOI":"10.1186\/gb-2009-10-6-r64","article-title":"A novel and universal method for microRNA RT-qPCR data normalization","volume":"10","author":"Mestdagh","year":"2009","journal-title":"Genome Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"5245","DOI":"10.1158\/0008-5472.CAN-04-0496","article-title":"Normalization of real-time quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets","volume":"64","author":"Andersen","year":"2004","journal-title":"Cancer Res."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Zhang, X., Goncalves, R., and Mosser, D.M. (2008). The isolation and characterization of murine macrophages. Curr. Protoc. Immunol.","DOI":"10.1002\/0471142735.im1401s83"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1021","DOI":"10.1128\/CVI.00017-11","article-title":"Kinetics and magnitude of antibody responses against the conserved 47-kilodalton antigen and the variable 56-kilodalton antigen in scrub typhus patients","volume":"18","author":"Chen","year":"2011","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1038\/ncomms1285","article-title":"Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells","volume":"2","author":"Mittelbrunn","year":"2011","journal-title":"Nat. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"7321","DOI":"10.1038\/ncomms8321","article-title":"Exosome-delivered microRNAs modulate the inflammatory response to endotoxin","volume":"6","author":"Alexander","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1111\/2049-632X.12135","article-title":"The carrying pigeons of the cell: Exosomes and their role in infectious diseases caused by human pathogens","volume":"71","author":"Fleming","year":"2014","journal-title":"Pathog. Dis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"W460","DOI":"10.1093\/nar\/gkv403","article-title":"DIANA-miRPath v3.0: Deciphering microRNA function with experimental support","volume":"43","author":"Vlachos","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Teng, G.G., Wang, W.H., Dai, Y., Wang, S.J., Chu, Y.X., and Li, J. (2013). Let-7b is involved in the inflammation and immune responses associated with Helicobacter pylori infection by targeting Toll-like receptor 4. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0056709"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1977","DOI":"10.1038\/emboj.2011.94","article-title":"Analysis of the host microRNA response to Salmonella uncovers the control of major cytokines by the let-7 family","volume":"30","author":"Schulte","year":"2011","journal-title":"EMBO J."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1016\/j.chom.2015.01.007","article-title":"MicroRNA let-7 modulates the immune response to Mycobacterium tuberculosis infection via control of A20, an inhibitor of the NF-kappaB pathway","volume":"17","author":"Kumar","year":"2015","journal-title":"Cell Host Microbe"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1007\/s13238-015-0212-y","article-title":"Biogenesis and regulation of the let-7 miRNAs and their functional implications","volume":"7","author":"Lee","year":"2016","journal-title":"Protein Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"12416","DOI":"10.1073\/pnas.1811040116","article-title":"Dual mechanisms of posttranscriptional regulation of Tet2 by Let-7 microRNA in macrophages","volume":"116","author":"Jiang","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_32","first-page":"491","article-title":"Macrophage cytokines: Involvement in immunity and infectious diseases","volume":"5","author":"Descoteaux","year":"2014","journal-title":"Front. Immunol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3234","DOI":"10.1182\/blood-2007-03-079152","article-title":"Exosomes released from macrophages infected with intracellular pathogens stimulate a proinflammatory response in vitro and in vivo","volume":"110","author":"Bhatnagar","year":"2007","journal-title":"Blood"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"866","DOI":"10.3346\/jkms.2015.30.7.866","article-title":"Identification of Outer Membrane Vesicles Derived from Orientia tsutsugamushi","volume":"30","author":"Lee","year":"2015","journal-title":"J. Korean Med. Sci."},{"key":"ref_35","first-page":"1500","article-title":"Non-Exosomal and Exosomal Circulatory MicroRNAs: Which Are More Valid as Biomarkers?","volume":"10","author":"Shahidan","year":"2019","journal-title":"Front. Pharmacol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Palviainen, M., Saraswat, M., Varga, Z., Kitka, D., Neuvonen, M., Puhka, M., Joenvaara, S., Renkonen, R., Nieuwland, R., and Takatalo, M. (2020). Extracellular vesicles from human plasma and serum are carriers of extravesicular cargo-Implications for biomarker discovery. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0236439"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"32651","DOI":"10.1038\/srep32651","article-title":"Platelets confound the measurement of extracellular miRNA in archived plasma","volume":"6","author":"Mitchell","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1016\/j.tcb.2008.07.007","article-title":"The let-7 family of microRNAs","volume":"18","author":"Roush","year":"2008","journal-title":"Trends Cell Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/j.canlet.2018.07.027","article-title":"Recent findings regarding let-7 in immunity","volume":"434","author":"Jiang","year":"2018","journal-title":"Cancer Lett."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"6238","DOI":"10.4049\/jimmunol.1101196","article-title":"Differential regulation of the Let-7 family of microRNAs in CD4+ T cells alters IL-10 expression","volume":"188","author":"Swaminathan","year":"2012","journal-title":"J. Immunol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Chao, C.C., Yang, R., Zhang, Z., Belinskaya, T., Chan, C.T., Miller, S.A., Hammamieh, R., Jett, M., and Ching, W.M. (2020). Temporal analysis of mRNA expression profiles in Orientia infected C3HeB\/FeJ mouse. BMC Microbiol., 20.","DOI":"10.1186\/s12866-019-1684-3"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/1\/80\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:48:17Z","timestamp":1760179697000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/1\/80"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,12,31]]},"references-count":41,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,1]]}},"alternative-id":["microorganisms9010080"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms9010080","relation":{},"ISSN":["2076-2607"],"issn-type":[{"type":"electronic","value":"2076-2607"}],"subject":[],"published":{"date-parts":[[2020,12,31]]}}}