Analysis of the Immune Responses in the Ileum of Gnotobiotic Pigs Infected with the Recombinant GII.p12_GII.3 Human Norovirus by mRNA Sequencing
Abstract
:1. Introduction
2. Materials and Methods
2.1. Virus Preparation
2.2. Animals and Experimental Design
2.3. Preparation of Fecal and Serum Samples
2.4. Detection of NoV in Fecal, Serum, and Tissue Samples
2.5. Determination of Viral Dose and mRNA Expression Levels
2.6. RNA Isolation for High-Throughput mRNA-Sequencing (RNA-Seq)
2.7. Library Preparation and Sequencing
2.8. Data Analysis
2.9. In Situ Hybridization for Histological Pathology and Western Blot Analysis
3. Results
3.1. Identification of HuNoV Infection
3.2. Inflammation and IFN Immune Responses
3.3. DC and NK Cell Activation
3.4. CD4+ and CD8+ T Cell-Mediated Adaptive Immune Responses
3.5. Evaluation of mRNA Sequencing Data by qRT-PCR and Western Blot
4. Discussion
4.1. Pathogenesis of HuNoV
4.2. Inflammatory Response
4.3. Innate Immune Response
4.4. NK Cell Activation
4.5. Expression of MHC Molecules and Cytokines
4.6. CD8+ T Cell Exhaustion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Group | dpi * | 0 | 1 | 2 | 3 | Jejunum | Ileum | Large Intestine | Mesenteric Lymph Node | Spleen | Liver |
---|---|---|---|---|---|---|---|---|---|---|---|
No. | F/S a | F/S | F/S | F/S | |||||||
Negative control | 428 | −/− | −/− | −/− | −/− | − | − | − | − | − | − |
429 | −/− | −/− | −/− | −/− | − | − | − | − | − | − | |
490 | −/− | −/− | −/− | −/− | − | − | − | − | − | − | |
404 | −/− | −/− | −/− | −/− | − | − | − | − | − | − | |
403 | −/− | −/− | −/− | −/− | − | − | − | − | − | − | |
HuNoV | 422 | −/− | −/− | +/− | +/+ | + | ++ | + | +++ | +++ | − |
423 | −/− | +/+ | +/+ | +/− | ++ | +++ | + | +++ | +++ | − | |
424 | −/− | +/− | +/+ | +− | + | +++ | + | +++ | +++ | − | |
497 | −/− | −/− | +/− | +/− | ++ | ++ | + | +++ | ++ | − | |
499 | −/− | +/− | +/− | −/− | + | +++ | + | +++ | + | − | |
425 | −/− | +/− | +/− | −/− | + | ++ | − | + | +++ | − | |
426 | −/− | +/− | +/− | +/− | + | ++ | + | + | +++ | − | |
427 | −/− | +/− | +/+ | +− | + | ++ | − | ++ | ++ | − | |
498 | −/− | −/− | +/− | +/− | − | + | + | + | + | − | |
500 | −/− | +/+ | +/+ | +− | + | +++ | + | +++ | ++ | − |
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Park, B.-J.; Ahn, H.-S.; Han, S.-H.; Go, H.-J.; Kim, D.-H.; Choi, C.; Jung, S.; Myoung, J.; Lee, J.-B.; Park, S.-Y.; et al. Analysis of the Immune Responses in the Ileum of Gnotobiotic Pigs Infected with the Recombinant GII.p12_GII.3 Human Norovirus by mRNA Sequencing. Viruses 2021, 13, 92. https://doi.org/10.3390/v13010092
Park B-J, Ahn H-S, Han S-H, Go H-J, Kim D-H, Choi C, Jung S, Myoung J, Lee J-B, Park S-Y, et al. Analysis of the Immune Responses in the Ileum of Gnotobiotic Pigs Infected with the Recombinant GII.p12_GII.3 Human Norovirus by mRNA Sequencing. Viruses. 2021; 13(1):92. https://doi.org/10.3390/v13010092
Chicago/Turabian StylePark, Byung-Joo, Hee-Seop Ahn, Sang-Hoon Han, Hyeon-Jeong Go, Dong-Hwi Kim, Changsun Choi, Soontag Jung, Jinjong Myoung, Joong-Bok Lee, Seung-Yong Park, and et al. 2021. "Analysis of the Immune Responses in the Ileum of Gnotobiotic Pigs Infected with the Recombinant GII.p12_GII.3 Human Norovirus by mRNA Sequencing" Viruses 13, no. 1: 92. https://doi.org/10.3390/v13010092