|
| Status |
Public on Jun 16, 2020 |
| Title |
Viral transcriptomic analysis of single cell-sorted of DF1 or MDCK cell lines infected with influenza A virus |
| Organisms |
Gallus gallus; Canis lupus familiaris |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Infection with a single influenza A virus (IAV) is only rarely sufficient to initiate productive infection. Here, we exploit both single cell approaches and whole-animal systems to show that the extent of IAV reliance on multiple infection varies with virus strain and host species. Influenza A/guinea fowl/HK/WF10/99 (H9N2) [GFHK99] virus exhibits strong dependence on collective interactions in mammalian systems. This reliance focuses viral progeny production within coinfected cells and therefore results in frequent genetic exchange through reassortment. In contrast, GFHK99 virus has greatly reduced dependence on multiple infection in avian systems, indicating a role for host factors in viral collective interactions. Genetic mapping implicated the viral polymerase as a major driver of multiple infection dependence. Mechanistically, quantification of incomplete viral genomes showed that their complementation only partly accounts for the observed reliance on coinfection. Indeed, even when all polymerase components are detected in single cell mRNA sequencing, robust polymerase activity of GFHK99 virus in mammalian cells is reliant on multiple infection. In sum, IAV collective interactions not only augment reassortment, but can also overcome species-specific barriers to infection. These findings underscore the importance of virus-virus interactions in IAV infection, evolution and emergence. We used a single-cell sequencing platform (10x Genomics) to elucidate the differential infection rate of an avian influenza A virus on an avian cell line (DF1) and a mammalian (MDCK) cell line. Our work on IAV reassortment has raised new questions about the fundamental strategies that drive influenza virus evolution. Our data indicate that a large majority of influenza virus genomesare incomplete within cells, comprising less than the eight complete segments normally found in a replication competent infectious viral particle. This led us to ask: what underlying mechanisms give rise to incomplete genomes? What constitute an infectious unit? What are the implications for viral diversification, evolution and spread. By addressing these questions, we will advance he field by deepening our understanding how viral infections are initiated and propagated.
|
| |
|
| Overall design |
Avian DF-1 cells or mammalian MDCK cells were infected with different multiplicity of infection using wildtype A/Guinea Fowl/Hong Kong/WF10/99 and barcoded A/Guinea Fowl/Hong Kong/WF10/99. At 8 hours post infection, cells were harvested for both experiments and processed for single-cell sorting and 3' transcript sequencing using the 10x Genomics platform.
|
| |
|
| Contributor(s) |
Phipps K, Ganti K, Carnaccini S, Manandhar M, Jacobs NT, Pickett BE, Tan GS, Ferreri LM, Perez DR, Lowen AC |
| Citation(s) |
32632248 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 AI127799 |
Host dependence of influenza A virus reassortment |
EMORY UNIVERSITY |
Anice C Lowen |
| U19 AI110819 |
The J. Craig Venter Institute Genome Center for Infectious Diseases |
J. Craig Venter Institute |
William Charles Nierman |
|
| |
| Submission date |
Aug 08, 2019 |
| Last update date |
Sep 15, 2020 |
| Contact name |
Gene Tan |
| E-mail(s) |
gtan@jcvi.org
|
| Organization name |
J. Craig Venter Institute
|
| Street address |
4120 Capricorn Lane
|
| City |
La Joll |
| ZIP/Postal code |
92037 |
| Country |
USA |
| |
|
| Platforms (2) |
| GPL19787 |
Illumina NextSeq 500 (Gallus gallus) |
| GPL21400 |
Illumina NextSeq 500 (Canis lupus familiaris) |
|
| Samples (16)
|
|
| Relations |
| BioProject |
PRJNA559274 |
| SRA |
SRP217802 |
Less...
More...