Mongolia Gerbils Are Broadly Susceptible to Hepatitis E Virus
<p>HEV replication in Mongolia gerbils. Twenty-seven Mongolia gerbils were randomly separated into nine groups (<span class="html-italic">n</span> = 3 per group). Individual gerbils are indicated by ○, △ and ☐ (<b>a</b>,<b>b</b>) or white, gray and black bars (<b>c</b>). Each group received HEV-1, HEV-3, HEV-4, HEV-5, HEV-7, HEV-8, rabbit HEV, rat HEV or phosphate-buffered saline (PBS) via intraperitoneal injection. The kinetics of the viral RNA in the fecal specimens were measured by RT-qPCR (<b>a</b>). The serum and tissue samples were collected at the end of the experiment (day 28 p.i.), and the anti-HEV-IgG antibody titers were determined using an ELISA with the VLPs of HEV-1 (to detect HEV-1, HEV-3, HEV-4, HEV-5, HEV-7, HEV-8 or rabbit HEV) or those of rat HEV (to detect rat HEV) as the antigens (<b>b</b>). Dotted lines: the cut-off values. The minimum endpoints of the antibody titers are blackened (●, ▲, ■) (<b>b</b>). The viral RNA titers in the liver, spleen and serum samples (<b>c</b>).</p> "> Figure 1 Cont.
<p>HEV replication in Mongolia gerbils. Twenty-seven Mongolia gerbils were randomly separated into nine groups (<span class="html-italic">n</span> = 3 per group). Individual gerbils are indicated by ○, △ and ☐ (<b>a</b>,<b>b</b>) or white, gray and black bars (<b>c</b>). Each group received HEV-1, HEV-3, HEV-4, HEV-5, HEV-7, HEV-8, rabbit HEV, rat HEV or phosphate-buffered saline (PBS) via intraperitoneal injection. The kinetics of the viral RNA in the fecal specimens were measured by RT-qPCR (<b>a</b>). The serum and tissue samples were collected at the end of the experiment (day 28 p.i.), and the anti-HEV-IgG antibody titers were determined using an ELISA with the VLPs of HEV-1 (to detect HEV-1, HEV-3, HEV-4, HEV-5, HEV-7, HEV-8 or rabbit HEV) or those of rat HEV (to detect rat HEV) as the antigens (<b>b</b>). Dotted lines: the cut-off values. The minimum endpoints of the antibody titers are blackened (●, ▲, ■) (<b>b</b>). The viral RNA titers in the liver, spleen and serum samples (<b>c</b>).</p> "> Figure 2
<p>Infectivity of HEV discharged into the feces. Three groups of Mongolia gerbils (<span class="html-italic">n</span> = 3 per group) were inoculated with the 10% stool suspension prepared from the HEV-4-inoculated, HEV-5-inoculated or rat HEV-inoculated gerbils. Individual gerbils are indicated by ○, △ and ☐ (<b>a</b>,<b>b</b>,<b>d</b>,<b>e</b>,<b>g</b>,<b>h</b>) or white, gray and black bars (<b>c</b>,<b>f</b>,<b>i</b>). The kinetics of the viral RNA in the fecal specimens were determined by RT-qPCR (<b>a</b>,<b>d</b>,<b>g</b>). The serum and tissue samples were collected at the end of the experiment (day 42 p.i.). The serum anti-HEV IgG antibodies were measured using an ELISA with either VLPs of HEV-1 (to detect HEV-4 and HEV-5) or those of rat HEV (to detect rat HEV) as the antigens. Dotted lines: the cut-off values and the minimum endpoints of the antibody titers are blackened (●, ▲, ■) (<b>b</b>,<b>e</b>,<b>h</b>). The viral RNA titers detected in the liver, spleen and serum (<b>c,f,i</b>).</p> "> Figure 3
<p>Detection of the viral RNA in the tissues in the early stage of infection. Six Mongolia gerbils were randomly separated into two groups (<span class="html-italic">n</span> = 3 per group). Individual gerbils are indicated by white, gray and black bars. One group was inoculated with HEV-4 (<b>a</b>) and the other with rat HEV (<b>b</b>) via intraperitoneal injection. All gerbils were euthanized on day 14 p.i. The serum samples, intestinal contents, liver, spleen, heart, lung, kidney, pancreas, salivary gland, thymus, muscle and brain were collected, and the viral RNA was measured by RT-qPCR.</p> "> Figure 4
<p>HEV-4, HEV-5 and rat HEV orally transmitted to Mongolia gerbils. Nine Mongolia gerbils were randomly separated into three groups (<span class="html-italic">n</span> = 3 per group). Individual gerbils are indicated by ○, △ and ☐ (<b>a</b>,<b>b</b>,<b>d</b>,<b>e</b>,<b>g</b>,<b>h</b>) or white, gray and black bars (<b>c</b>,<b>f</b>,<b>i</b>). Each group was orally inoculated with HEV-4, HEV-5 or rat HEV. The kinetics of the viral RNA in the fecal specimens were determined by RT-qPCR (<b>a</b>,<b>d</b>,<b>g</b>). The liver, spleen, bile and serum samples were collected at the end of the experiment (day 26 p.i.). The serum anti-HEV IgG antibodies were detected by using an ELISA with either the VLPs of HEV-1 (to detect HEV-4 and HEV-5) or those of rat HEV (to detect rat HEV) as the antigens. Dotted lines: the cut-off value and the minimum endpoints of the antibody titers were blackened (●, ▲, ■) (<b>b</b>,<b>e</b>,<b>h</b>). The viral RNA titers in the liver, spleen, bile and serum (<b>c</b>,<b>f</b>,<b>i</b>).</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. HEV Strains
2.2. Inoculation of Mongolia Gerbils and the Sample Collection
2.3. Extraction and Detection of HEV RNA
2.4. Detection of Anti-HEV IgG Antibodies
2.5. Liver Enzyme Level
2.6. Viral Genome Sequencing
3. Results
3.1. Mongolia Gerbils Had Different Susceptibilities to HEV
3.2. Analyses of the Entire Genomes of HEV-4, HEV-5, and Rat HEV
3.3. Infectivity of HEV Excreted in the Fecal Specimens
3.4. Detection of HEV in Tissues at the Early Stage of Infection
3.5. Transmission of HEV-4, HEV-5, and Rat HEV by an Oral Inoculation
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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HEV Strain | Gerbil 1 | Gerbil 2 | Gerbil 3 |
---|---|---|---|
HEV-1 | 45 | 66 | 47 |
HEV-3 | 54 | 49 | 69 |
HEV-4 | 63 | 69 | 78 |
HEV-5 | 59 | 58 | 78 |
HEV-7 | 46 | 46 | 63 |
HEV-8 | 54 | 66 | 60 |
Rabbit HEV | 42 | 47 | 48 |
Rat HEV | 57 | 62 | 59 |
PBS | 53 | 57 | 60 |
HEV Strain | Gerbil 1 | Gerbil 2 | Gerbil 3 |
---|---|---|---|
HEV-4 | 59 | 58 | 98 |
Rat HEV | 92 | 70 | 61 |
HEV Strain | Gerbil 1 | Gerbil 2 | Gerbil 3 |
---|---|---|---|
HEV-4 | 53 | 47 | 56 |
HEV-5 | 56 | 70 | 61 |
Rat HEV | 56 | 50 | 59 |
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Zhang, W.; Ami, Y.; Suzaki, Y.; Doan, Y.H.; Muramatsu, M.; Li, T.-C. Mongolia Gerbils Are Broadly Susceptible to Hepatitis E Virus. Viruses 2022, 14, 1125. https://doi.org/10.3390/v14061125
Zhang W, Ami Y, Suzaki Y, Doan YH, Muramatsu M, Li T-C. Mongolia Gerbils Are Broadly Susceptible to Hepatitis E Virus. Viruses. 2022; 14(6):1125. https://doi.org/10.3390/v14061125
Chicago/Turabian StyleZhang, Wenjing, Yasushi Ami, Yuriko Suzaki, Yen Hai Doan, Masamichi Muramatsu, and Tian-Cheng Li. 2022. "Mongolia Gerbils Are Broadly Susceptible to Hepatitis E Virus" Viruses 14, no. 6: 1125. https://doi.org/10.3390/v14061125