The Vector Competence of Asian Longhorned Ticks in Langat Virus Transmission
<p>Replication of LGTV in <span class="html-italic">H. longicornis</span> nymphs through microinjection. (<b>A</b>) Cell image of LGTV-BHK21 cell. (<b>B</b>) The supernatant of LGTV-BHK21 cells was collected. Viral particles are serially diluted and added onto confluent cell monolayers of BHK21, and the plaque assay was performed to determine viral titers. (<b>C</b>,<b>D</b>) <span class="html-italic">H. longicornis</span> nymphs were injected with LGTV via anal pore (<b>C</b>) and hemolymph injection (<b>D</b>).Viral RNA was determined using RT-qPCR at the indicated time points. The expression level of <span class="html-italic">preM</span> was normalized to actin. The relative expression levels of <span class="html-italic">preM</span> in ticks from 14 days post injection (D14) to 42 days post injection (D42) were normalized to those in ticks one day post injection (D1). Three nymphs were pooled for one biological replicate. Each dot represented a biological replicate. (<b>E</b>) Schematic depiction of the experimental design. Six-to-twelve-week-old A6 mice were infected intraperitoneally with 10 pfu LGTV. The nymphs were allowed to bite the infected A6 mice. LGTV was quantified in nymphs that had been feeding for two days (<b>F</b>) or four days (<b>G</b>). Two nymphs were pooled for one biological replicate in (<b>F</b>), and an individual engorgement nymph was one biological replicate in (<b>G</b>). Each dot represents a biological replicate. Significance was determined using Student’s <span class="html-italic">t</span> test. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001, **** <span class="html-italic">p</span> < 0.0001.</p> "> Figure 2
<p>LGTV is detectable in various tick organs. (<b>A</b>–<b>G</b>) Viral RNA from midguts (<b>A</b>,<b>D</b>), salivary glands (<b>B</b>,<b>E</b>), synganglion (<b>C</b>,<b>F</b>), and ovaries (<b>G</b>) of engorgement nymphs (<b>A</b>–<b>C</b>) and adult ticks (<b>D</b>–<b>G</b>) were quantified using RT-qPCR. Three midguts, three salivary glands, three synganglion, or three ovaries were pooled for one biological replicate. Each dot represents a biological replicate. (<b>H</b>) Tissue localization of the LGTV (red) in infected ticks from adult ticks. Viral antigens were detected using a specific LGTV anti-EDⅢ polyclonal antibody, while the organ from normal ticks which bit DMEM-injected A6 mice served as a control. Nuclei counterstaining (blue) was performed using DAPI. Dashed boxes denote LGTV. Images were representative of 10 ticks. Scale bars, 25 µm. MG, midgut; SG, salivary glands; SYN, synganglion; OV, ovary.</p> "> Figure 3
<p>LGTV is transstadially but not transovarially transmitted in <span class="html-italic">H. longicornis</span>. (<b>A</b>) Schematic depiction of the experimental design. Six-to-twelve-week-old A6 mice were infected intraperitoneally with 10 pfu LGTV, and the larvae or nymphs were allowed to bite the infected A6 mice. The engorged larvae or nymphs were collected and left to molt to the next stage. Viral RNA was quantified using qRT-PCR. (<b>B</b>,<b>C</b>) The viral RNA of the next stage, nymphs or adults, was quantified. Each dot represents three nymphs or one adult tick. (<b>D</b>) Schematic representation of the experimental design. Adult ticks were injected with 4000 pfu LGTV and the infected adult ticks were allowed to bite <span class="html-italic">BALB/C</span> to engorgement. The engorged adult ticks laid eggs and molted to larvae. (<b>E</b>) The infection of adult ticks was detected at 1 dpi. Each dot represents an adult tick. (<b>F</b>) RT-qPCR was performed to determine the viral RNA of larvae. A total of 50 larvae were pooled for one biological replicate. Each dot represents one biological replicate.</p> "> Figure 4
<p>LGTV is transmitted from infected ticks to A6 mice or naïve nymphs. (<b>A</b>) The nymphs obtained LGTV from infected A6 mice. The engorged nymphs were collected (Day 0 ticks) and after 14 days had molted to adult ticks. It would take 28 days for adults to become hungry and ready to take a blood meal (Day 28). Viral RNA was detected using qRT-PCR at the appropriate time points. Each dot represents an individual tick. (<b>B</b>) The plaque assay was performed to determine viral titers within the supernatant of infected adults 28 days post molting to adults. (<b>C</b>) Transmission from LGTV-infected adult ticks to susceptible A6 mice. LGTV-infected ticks (Day 28) were allowed to bite A6 mice, one tick per A6 mouse. The viral RNA of the blood-fed-upon A6 mice was determined three days post feeding. Each dot represents a blood sample collected from one A6 mouse.</p> "> Figure 5
<p>LGTV is transmitted from infected ticks to naïve nymphs through horizontal transmission. (<b>A</b>) Schematic depiction of the experimental design. A total of 15 nymphs were injected with 150 pfu LGTV via anal pore microinjection (red circle), 15 nymphs injected with 15 nL DMEM medium served as a control. A total of 30 treated nymphs were allowed to co-feed on one A6 mouse until engorgement and allowed to molt to an adult tick. Viral RNA was extracted and determined using RT-qPCR. (<b>B</b>) Blood samples were collected when the nymphs were engorged.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Tick Maintenance
2.2. Cell Culture and Virus Amplification
2.3. Plaque Assays
2.4. Tick Infection
2.5. Mice Infection
2.6. RNA Extraction and Quantitative Real-Time PCR (qRT-PCR)
2.7. Transstadial Transmission Analysis of LGTV
2.8. Transovarial Transmission Analysis of LGTV
2.9. Horizontal Transmission Analysis of LGTV
2.10. LGTV Transmission Assay between Ticks and Mice
2.11. Antibodies and Western Blot
2.12. Immunohistochemistry
2.13. Statistical Analysis
3. Results
3.1. Replication of LGTV in H. longicornis
3.2. The Tissue Tropism of LGTV in H. longicornis
3.3. Transstadial and Transovarial Transmission of LGTV in H. longicornis
3.4. Transmission of LGTV from H. longicornis to Mice
3.5. Horizontal Transmission of LGTV among Ticks during Blood Feeding
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Raney, W.R.; Herslebs, E.J.; Langohr, I.M.; Stone, M.C.; Hermance, M.E. Horizontal and Vertical Transmission of Powassan Virus by the Invasive Asian Longhorned Tick, Haemaphysalis longicornis, Under Laboratory Conditions. Front. Cell Infect. Microbiol. 2022, 12, 923914. [Google Scholar] [CrossRef]
- Bartikova, P.; Holikova, V.; Kazimirova, M.; Stibraniova, I. Tick-borne viruses. Acta Virol. 2017, 61, 413–427. [Google Scholar] [CrossRef]
- Yu, X.J.; Liang, M.F.; Zhang, S.Y.; Liu, Y.; Li, J.D.; Sun, Y.L.; Zhang, L.; Zhang, Q.F.; Popov, V.L.; Li, C.; et al. Fever with thrombocytopenia associated with a novel bunyavirus in China. N. Engl. J. Med. 2011, 364, 1523–1532. [Google Scholar] [CrossRef]
- Kim, K.H.; Yi, J.; Kim, G.; Choi, S.J.; Jun, K.I.; Kim, N.H.; Choe, P.G.; Kim, N.J.; Lee, J.K.; Oh, M.D. Severe fever with thrombocytopenia syndrome, South Korea, 2012. Emerg. Infect. Dis. 2013, 19, 1892–1894. [Google Scholar] [CrossRef]
- Li, S.; Li, H.; Zhang, Y.L.; Xin, Q.L.; Guan, Z.Q.; Chen, X.; Zhang, X.A.; Li, X.K.; Xiao, G.F.; Lozach, P.Y.; et al. SFTSV Infection Induces BAK/BAX-Dependent Mitochondrial DNA Release to Trigger NLRP3 Inflammasome Activation. Cell Rep. 2020, 30, 4370–4385.e4377. [Google Scholar] [CrossRef] [PubMed]
- Yoshii, K. Epidemiology and pathological mechanisms of tick-borne encephalitis. J. Vet. Med. Sci. 2019, 81, 343–347. [Google Scholar] [CrossRef] [PubMed]
- Gritsun, T.S.; Lashkevich, V.A.; Gould, E.A. Tick-borne encephalitis. Antivir. Res. 2003, 57, 129–146. [Google Scholar] [CrossRef] [PubMed]
- Pulkkinen, L.I.A.; Butcher, S.J.; Anastasina, M. Tick-Borne Encephalitis Virus: A Structural View. Viruses 2018, 10, 350. [Google Scholar] [CrossRef] [PubMed]
- Charrel, R.N.; Zaki, A.M.; Attoui, H.; Fakeeh, M.; Billoir, F.; Yousef, A.I.; de Chesse, R.; De Micco, P.; Gould, E.A.; de Lamballerie, X. Complete coding sequence of the Alkhurma virus, a tick-borne flavivirus causing severe hemorrhagic fever in humans in Saudi Arabia. Biochem. Biophys. Res. Commun. 2001, 287, 455–461. [Google Scholar] [CrossRef]
- Talactac, M.R.; Yoshii, K.; Maeda, H.; Kusakisako, K.; Hernandez, E.P.; Tsuji, N.; Fujisaki, K.; Galay, R.L.; Tanaka, T.; Mochizuki, M. Virucidal activity of Haemaphysalis longicornis longicin P4 peptide against tick-borne encephalitis virus surrogate Langat virus. Parasit Vectors 2016, 9, 59. [Google Scholar] [CrossRef] [PubMed]
- Kirsch, J.M.; Mlera, L.; Offerdahl, D.K.; VanSickle, M.; Bloom, M.E. Tick-Borne Flaviviruses Depress AKT Activity during Acute Infection by Modulating AKT1/2. Viruses 2020, 12, 1059. [Google Scholar] [CrossRef] [PubMed]
- Maffioli, C.; Grandgirard, D.; Olivier, E.; Leib, S.L. A Tick-Borne Encephalitis Model in Infant Rats Infected. J. Neuropathol. Exp. Neurol. 2014, 73, 1107–1115. [Google Scholar] [CrossRef]
- Schreier, S.; Cebulski, K.; Kröger, A. Contact-Dependent Transmission of Langat and Tick-Borne. J. Virol. 2021, 95, 10–1128. [Google Scholar] [CrossRef]
- Hernandez, E.P.; Talactac, M.R.; Vitor, R.J.S.; Yoshii, K.; Tanaka, T. An Ixodes scapularis glutathione S-transferase plays a role in cell survival and viability during Langat virus infection of a tick cell line. Acta Trop. 2021, 214, 105763. [Google Scholar] [CrossRef] [PubMed]
- Kusakisako, K.; Morokuma, H.; Talactac, M.R.; Hernandez, E.P.; Yoshii, K.; Tanaka, T. A Peroxiredoxin from the Haemaphysalis longicornis Tick Affects Langat Virus Replication in a Hamster Cell Line. Front. Cell Infect. Microbiol. 2020, 10, 7. [Google Scholar] [CrossRef] [PubMed]
- Lewy, T.G.; Offerdahl, D.K.; Grabowski, J.M.; Kellman, E.; Mlera, L.; Chiramel, A.; Bloom, M.E. PERK-Mediated Unfolded Protein Response Signaling Restricts Replication of the Tick-Borne Flavivirus Langat Virus. Viruses 2020, 12, 328. [Google Scholar] [CrossRef]
- Mlera, L.; Melik, W.; Offerdahl, D.K.; Dahlstrom, E.; Porcella, S.F.; Bloom, M.E. Analysis of the Langat Virus Genome in Persistent Infection of an Ixodes scapularis Cell Line. Viruses 2016, 8, 252. [Google Scholar] [CrossRef]
- Zhong, Z.; Zhong, T.; Peng, Y.; Zhou, X.; Wang, Z.; Tang, H.; Wang, J. Symbiont-regulated serotonin biosynthesis modulates tick feeding activity. Cell Host Microbe 2021, 29, 1545–1557.e4. [Google Scholar] [CrossRef]
- Talactac, M.R.; Yoshii, K.; Hernandez, E.P.; Kusakisako, K.; Galay, R.L.; Fujisaki, K.; Mochizuki, M.; Tanaka, T. Synchronous Langat Virus Infection of Haemaphysalis longicornis Using Anal Pore Microinjection. Viruses 2017, 9, 189. [Google Scholar] [CrossRef]
- Yu, X.; Shan, C.; Zhu, Y.; Ma, E.; Wang, J.; Wang, P.; Shi, P.Y.; Cheng, G. A mutation-mediated evolutionary adaptation of Zika virus in mosquito and mammalian host. Proc. Natl. Acad. Sci. USA 2021, 118, e2113015118. [Google Scholar] [CrossRef]
- Zhang, Y.; Xu, Y.; Dai, Y.; Li, Z.; Wang, J.; Ye, Z.; Ren, Y.; Wang, H.; Li, W.X.; Lu, J.; et al. Efficient Dicer processing of virus-derived double-stranded RNAs and its modulation by RIG-I-like receptor LGP2. PLoS Pathog. 2021, 17, e1009790. [Google Scholar] [CrossRef]
- Talactac, M.R.; Yoshii, K.; Hernandez, E.P.; Kusakisako, K.; Galay, R.L.; Fujisaki, K.; Mochizuki, M.; Tanaka, T. Vector competence of Haemaphysalis longicornis ticks for a Japanese isolate of the Thogoto virus. Sci. Rep. 2018, 8, 9300. [Google Scholar] [CrossRef]
- Weber, E.; Finsterbusch, K.; Lindquist, R.; Nair, S.; Lienenklaus, S.; Gekara, N.O.; Janik, D.; Weiss, S.; Kalinke, U.; Overby, A.K.; et al. Type I interferon protects mice from fatal neurotropic infection with Langat virus by systemic and local antiviral responses. J. Virol. 2014, 88, 12202–12212. [Google Scholar] [CrossRef]
- Mukherjee, M.; Dutta, K.; White, M.A.; Cowburn, D.; Fox, R.O. NMR solution structure and backbone dynamics of domain III of the E protein of tick-borne Langat flavivirus suggests a potential site for molecular recognition. Protein Sci. 2006, 15, 1342–1355. [Google Scholar] [CrossRef] [PubMed]
- Gao, L.; Song, X.; Wang, J. Gut microbiota is essential in PGRP-LA regulated immune protection against Plasmodium berghei infection. Parasit Vectors 2020, 13, 3. [Google Scholar] [CrossRef]
- Tang, X.; Cao, Y.; Arora, G.; Hwang, J.; Sajid, A.; Brown, C.L.; Mehta, S.; Marin-Lopez, A.; Chuang, Y.M.; Wu, M.J.; et al. The Lyme disease agent co-opts adiponectin receptor-mediated signaling in its arthropod vector. eLife 2021, 10, e72568. [Google Scholar] [CrossRef]
- Maqbool, M.; Sajid, M.S.; Saqib, M.; Anjum, F.R.; Tayyab, M.H.; Rizwan, H.M.; Rashid, M.I.; Rashid, I.; Iqbal, A.; Siddique, R.M.; et al. Potential Mechanisms of Transmission of Tick-Borne Viruses at the Virus-Tick Interface. Front. Microbiol. 2022, 13, 846884. [Google Scholar] [CrossRef] [PubMed]
- Luo, L.M.; Zhao, L.; Wen, H.L.; Zhang, Z.T.; Liu, J.W.; Fang, L.Z.; Xue, Z.F.; Ma, D.Q.; Zhang, X.S.; Ding, S.J.; et al. Haemaphysalis longicornis Ticks as Reservoir and Vector of Severe Fever with Thrombocytopenia Syndrome Virus in China. Emerg. Infect. Dis. 2015, 21, 1770–1776. [Google Scholar] [CrossRef]
- Ferreira, F.C.; Gonzalez, J.; Milholland, M.T.; Tung, G.A.; Fonseca, D.M. Ticks (Acari: Ixodida) on synanthropic small and medium-sized mammals in areas of the northeastern United States infested with the Asian longhorned tick, Haemaphysalis longicornis. Int. J. Parasitol. 2023, 53, 809–819. [Google Scholar] [CrossRef]
- Jia, N.; Wang, J.; Shi, W.; Du, L.; Ye, R.Z.; Zhao, F.; Cao, W.C. Haemaphysalis longicornis. Trends Genet. 2021, 37, 292–293. [Google Scholar] [CrossRef] [PubMed]
- Ahmed, W.; Rajendran, K.V.; Neelakanta, G.; Sultana, H. An Experimental Murine Model to Study Acquisition Dynamics of Tick-Borne Langat Virus in Ixodes scapularis. Front. Microbiol. 2022, 13, 849313. [Google Scholar] [CrossRef] [PubMed]
- Hajdusek, O.; Sima, R.; Ayllon, N.; Jalovecka, M.; Perner, J.; de la Fuente, J.; Kopacek, P. Interaction of the tick immune system with transmitted pathogens. Front. Cell Infect. Microbiol. 2013, 3, 26. [Google Scholar] [CrossRef]
- Schafer, M.; Pfaff, F.; Hoper, D.; Silaghi, C. Early Transcriptional Changes in the Midgut of Ornithodoros moubata after Feeding and Infection with Borrelia duttonii. Microorganisms 2022, 10, 525. [Google Scholar] [CrossRef]
- Fogaca, A.C.; Sousa, G.; Pavanelo, D.B.; Esteves, E.; Martins, L.A.; Urbanova, V.; Kopacek, P.; Daffre, S. Tick Immune System: What Is Known, the Interconnections, the Gaps, and the Challenges. Front. Immunol. 2021, 12, 628054. [Google Scholar] [CrossRef] [PubMed]
- Lejal, E.; Moutailler, S.; Šimo, L.; Vayssier-Taussat, M.; Pollet, T. Tick-borne pathogen detection in midgut and salivary glands of adult Ixodes ricinus. Parasites Vectors 2019, 12, 152. [Google Scholar] [CrossRef]
- Piesman, J.; Schneider, B.S. Dynamic changes in Lyme disease spirochetes during transmission by nymphal ticks. Exp. Appl. Acarol. 2002, 28, 141–145. [Google Scholar] [CrossRef]
- Waldman, J.; Klafke, G.M.; Tirloni, L.; Logullo, C.; da Silva Vaz, I., Jr. Putative target sites in synganglion for novel ixodid tick control strategies. Ticks Tick Borne Dis. 2023, 14, 102123. [Google Scholar] [CrossRef]
- Grabowski, J.M.; Tsetsarkin, K.A.; Long, D.; Scott, D.P.; Rosenke, R.; Schwan, T.G.; Mlera, L.; Offerdahl, D.K.; Pletnev, A.G.; Bloom, M.E. Flavivirus Infection of Ixodes scapularis (Black-Legged Tick) Ex Vivo Organotypic Cultures and Applications for Disease Control. mBio 2017, 8, 10–1128. [Google Scholar] [CrossRef] [PubMed]
- Raney, W.R.; Perry, J.B.; Hermance, M.E. Transovarial Transmission of Heartland Virus by Invasive Asian Longhorned Ticks under Laboratory Conditions. Emerg. Infect. Dis. 2022, 28, 726–729. [Google Scholar] [CrossRef]
- Yuan, C.; Lu, Y.; Li, J.; Chen, C.; Wang, Y.; Zheng, A.; Zou, Z.; Xia, Q. Infection and transovarial transmission of severe fever with thrombocytopenia syndrome virus in Rhipicephalus sanguineus in Hainan Island, China. Integr. Zool. 2023, 18, 1009–1013. [Google Scholar] [CrossRef]
- Bartikova, P.; Stibraniova, I.; Kazimirova, M. Discovery of the Role of Tick Salivary Glands in Enhancement of Virus Transmission-Beginning of an Exciting Story. Pathogens 2023, 12, 334. [Google Scholar] [CrossRef] [PubMed]
- Jones, L.D.; Davies, C.R.; Steele, G.M.; Nuttall, P.A. A Novel Mode of Arbovirus Transmission Involving a Nonviremic Host. Science 1987, 237, 775–777. [Google Scholar] [CrossRef] [PubMed]
Treatment: LGTV/DMEM-Injected Nymphs | LGTV Detection | Percentage% (Positive/Total) |
---|---|---|
15 + 15 (Exp1.) | Engorged nymphs | 78.6% (22/28) |
15 + 15 (Exp2.) | Engorged nymphs | 96.7% (29/30) |
15 + 15 (Exp1.) | Adult ticks | 83.3% (20/24) |
15 + 15 (Exp2.) | Adult ticks | 80.8% (21/26) |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Xu, Y.; Wang, J. The Vector Competence of Asian Longhorned Ticks in Langat Virus Transmission. Viruses 2024, 16, 304. https://doi.org/10.3390/v16020304
Xu Y, Wang J. The Vector Competence of Asian Longhorned Ticks in Langat Virus Transmission. Viruses. 2024; 16(2):304. https://doi.org/10.3390/v16020304
Chicago/Turabian StyleXu, Yan, and Jingwen Wang. 2024. "The Vector Competence of Asian Longhorned Ticks in Langat Virus Transmission" Viruses 16, no. 2: 304. https://doi.org/10.3390/v16020304