Type I interferon production during herpes simplex virus infection is controlled by cell-type-specific viral recognition through Toll-like receptor 9, the mitochondrial antiviral signaling protein pathway, and novel recognition systems - PubMed (original) (raw)
Type I interferon production during herpes simplex virus infection is controlled by cell-type-specific viral recognition through Toll-like receptor 9, the mitochondrial antiviral signaling protein pathway, and novel recognition systems
Simon B Rasmussen et al. J Virol. 2007 Dec.
Abstract
Recognition of viruses by germ line-encoded pattern recognition receptors of the innate immune system is essential for rapid production of type I interferon (IFN) and early antiviral defense. We investigated the mechanisms of viral recognition governing production of type I IFN during herpes simplex virus (HSV) infection. We show that early production of IFN in vivo is mediated through Toll-like receptor 9 (TLR9) and plasmacytoid dendritic cells, whereas the subsequent alpha/beta IFN (IFN-alpha/beta) response is derived from several cell types and induced independently of TLR9. In conventional DCs, the IFN response occurred independently of viral replication but was dependent on viral entry. Moreover, using a HSV-1 UL15 mutant, which fails to package viral DNA into the virion, we found that entry-dependent IFN induction also required the presence of viral genomic DNA. In macrophages and fibroblasts, where the virus was able to replicate, HSV-induced IFN-alpha/beta production was dependent on both viral entry and replication, and ablated in cells unable to signal through the mitochondrial antiviral signaling protein pathway. Thus, during an HSV infection in vivo, multiple mechanisms of pathogen recognition are active, which operate in cell-type- and time-dependent manners to trigger expression of type I IFN and coordinate the antiviral response.
Figures
FIG. 1.
The pDC-TLR9 pathway is responsible for early but not for late IFN-α/β production during HSV infection in vivo. (A to H) C57BL/6 or TLR9−/− mice were infected i.p. with 2 × 107 PFU of HSV-1 or HSV-2 (five mice per group). At the indicated time points p.i., sera were harvested (A to F), or spleen cDCs and pDCs were isolated and cultured for ex vivo cytokine expression for 24 h (G to H). The levels of IFN-α/β were measured by bioassay. Similar results were obtained in three to four independent experiments. (I to L) WT mice (129sv in panels I and J; C57BL/6 in panels K and L), IFNAR−/− mice, and TLR9−/− mice were infected i.p. with 106 PFU of HSV-2 (five mice per group). At 2 days p.i., livers and spleens were harvested, and the viral loads in the organs were determined by plaque assay. Similar results were obtained in three independent experiments. (M) Mice treated as in panels I to L were monitored for 6 days. The mice were sacrificed when they displayed symptoms irreversibly associated with death. Error bars indicate the standard errors of the mean (SEM). *, P < 0.05.
FIG. 2.
Cell-type-specific requirements for TLR9 and virus replication for induction of IFN-α/β by HSV-2 in vitro. HSV-2 was treated with UV light for the indicated time intervals before being subjected to plaque assay (A) or used for infection of Vero cells for 5 h at an MOI of 1 (B) before harvesting of total RNA and detection of ICP27 and β-actin by RT-PCR. Similar results were obtained in two independent experiments. (C to J) Cells were harvested from WT or TLR9−/− mice and cultured with medium alone or in the presence of 3 × 106 PFU of HSV-2/ml or an equivalent amount of virus UV inactivated for 4 min. Supernatants were harvested 24 h p.i., and the levels of IFN-α/β were determined by bioassay. Similar results were obtained in three independent experiments. (K to N) Splenic pDCs, cDCs, and macrophages, as well as MEFs, were cultured and infected with 105 PFU of HSV-2/ml. Supernatants were harvested 6, 24, and 48 h p.i., and the viral load was measured by plaque assay. Similar results were obtained in two independent experiments. Error bars indicate the SEM.
FIG. 3.
IFN-α/β expression in vivo involves at least three mechanisms with differential requirements for TLR9 and virus replication. C57BL/6 or TLR9−/− mice were infected i.p. with 2 × 107 PFU of infectious or inactivated HSV-1 (A and C) or HSV-2 (B and D) (five mice per group). After 8 h (A and B) or 16 h (C and D) of treatment, serum was harvested, and the levels of IFN-α/β were measured by bioassay. (E and F) Splenic cDCs and macrophages were harvested from TLR9−/− mice infected as described above for 16 h. The cells were cultured for 24 h, and the ex vivo production of IFN was measured by bioassay. Similar results were obtained in three independent experiments. Error bars indicate the SEM. *, P < 0.05.
FIG. 4.
HSV-induced expression of IFN-α/β is dependent on virion DNA and displays a differential requirement for viral entry. (A) Thioglycolate-activated macrophages were harvested from BALB/c mice and treated with 3 × 106 PFU of HSV-1 or ΔgL/ml for 24 h. IL-12p40 was measured by ELISA. Similar results were obtained in three independent experiments. (B and C) Genomic DNA or total lysates of the preparations of the indicated viruses were analyzed for content of DNA and gD protein, respectively (upper panels). Nuclear extracts were prepared from Vero cells treated for 3 h with the indicated virus at an MOI of 12. VP16 was detected in the extracts by Western blotting (lower panel). Similar results were obtained in three independent experiments. (D to I) Splenic pDCs, cDCs, and macrophages, together with MEFs from C57BL/6 and TLR9−/− mice, were cultured and treated for 20 h with 3 × 106 PFU of HSV-1, ΔgL, or ΔUL15 viruses/ml as indicated. The supernatants were harvested, and the IFN-α/β levels were measured. Similar results were obtained in three to four independent experiments. (J) C57BL/6 and TLR9−/− mice were injected i.p. with 2 × 107 PFU of HSV-1, ΔgL, or ΔUL15 viruses (five mice per group). After 16 h, sera were harvested, and the type I IFN levels were measured. Similar results were obtained in three independent experiments. Error bars indicate the SEM.
FIG. 5.
MEFs use the MAVS pathway to induce type I IFN production in response to HSV infection. (A and B) MEFs from C57BL/6 or MAVS−/− mice were seeded and infected with 3 × 106 PFU of HSV-1 or HSV-2/ml. After 24 and 4 h, supernatants and total RNA were harvested for measurement of the IFN-α/β bioactivity (A) and mRNA (B), respectively. (C) MEFs from C57BL/6 or MAVS−/− mice were seeded and transfected with calf thymus-derived DNA using Lipofectamine 2000. Twenty-four hours later the supernatants were harvested for measurement of the IFN-α/β bioactivity. (D) MEFs from C57BL/6 mice were seeded and infected with 3 × 106 PFU/ml of the 17+ or KOS strains of HSV-1 and the indicated mutants. After 24 h, the supernatants were harvested, and the IFN-α/β levels were measured. For all data in this figure, similar results were obtained in three independent experiments. Error bars indicate the SEM.
Similar articles
- The virion host shutoff protein of herpes simplex virus 1 blocks the replication-independent activation of NF-κB in dendritic cells in the absence of type I interferon signaling.
Cotter CR, Kim WK, Nguyen ML, Yount JS, López CB, Blaho JA, Moran TM. Cotter CR, et al. J Virol. 2011 Dec;85(23):12662-72. doi: 10.1128/JVI.05557-11. Epub 2011 Sep 21. J Virol. 2011. PMID: 21937652 Free PMC article. - Herpes simplex virus type-1 induces IFN-alpha production via Toll-like receptor 9-dependent and -independent pathways.
Hochrein H, Schlatter B, O'Keeffe M, Wagner C, Schmitz F, Schiemann M, Bauer S, Suter M, Wagner H. Hochrein H, et al. Proc Natl Acad Sci U S A. 2004 Aug 3;101(31):11416-21. doi: 10.1073/pnas.0403555101. Epub 2004 Jul 22. Proc Natl Acad Sci U S A. 2004. PMID: 15272082 Free PMC article. - Herpes simplex virus type 2 virion host shutoff protein regulates alpha/beta interferon but not adaptive immune responses during primary infection in vivo.
Murphy JA, Duerst RJ, Smith TJ, Morrison LA. Murphy JA, et al. J Virol. 2003 Sep;77(17):9337-45. doi: 10.1128/jvi.77.17.9337-9345.2003. J Virol. 2003. PMID: 12915549 Free PMC article. - Herpes Simplex Virus Type 1 Interactions with the Interferon System.
Danastas K, Miranda-Saksena M, Cunningham AL. Danastas K, et al. Int J Mol Sci. 2020 Jul 21;21(14):5150. doi: 10.3390/ijms21145150. Int J Mol Sci. 2020. PMID: 32708188 Free PMC article. Review. - Herpes Simplex Virus 1 Infection of Neuronal and Non-Neuronal Cells Elicits Specific Innate Immune Responses and Immune Evasion Mechanisms.
Verzosa AL, McGeever LA, Bhark SJ, Delgado T, Salazar N, Sanchez EL. Verzosa AL, et al. Front Immunol. 2021 May 31;12:644664. doi: 10.3389/fimmu.2021.644664. eCollection 2021. Front Immunol. 2021. PMID: 34135889 Free PMC article. Review.
Cited by
- Clinical epidemiology, determinants, and outcomes of viral encephalitis in Ghana; a cross-sectional study.
Yeboah R, Gorman R, Acheampong HK, Nyarko-Afriyie E, Aryeetey S, Tetteh HD, Owusu M, Yeboah ES, Adade T, Bonney J, Amoako YA, El-Duah P, Obiri-Danso K, Drosten C, Phillips RO, Sylverken AA. Yeboah R, et al. PLoS One. 2024 Feb 12;19(2):e0297277. doi: 10.1371/journal.pone.0297277. eCollection 2024. PLoS One. 2024. PMID: 38346087 Free PMC article. - Cell Intrinsic Determinants of Alpha Herpesvirus Latency and Pathogenesis in the Nervous System.
Salazar S, Luong KTY, Koyuncu OO. Salazar S, et al. Viruses. 2023 Nov 22;15(12):2284. doi: 10.3390/v15122284. Viruses. 2023. PMID: 38140525 Free PMC article. Review. - Role of IL-27 in HSV-1-Induced Herpetic Stromal Keratitis.
Antony F, Pundkar C, Sandey M, Mishra A, Suryawanshi A. Antony F, et al. J Immunol. 2023 Aug 1;211(3):474-485. doi: 10.4049/jimmunol.2200420. J Immunol. 2023. PMID: 37326494 Free PMC article. - Intralesional administration of VAX014 facilitates in situ immunization and potentiates immune checkpoint blockade in immunologically cold tumors.
Reil KA, Tsuji S, Molina E, Nelson KL, McGuire KL, Giacalone MJ. Reil KA, et al. J Immunother Cancer. 2023 Jun;11(6):e006749. doi: 10.1136/jitc-2023-006749. J Immunother Cancer. 2023. PMID: 37290924 Free PMC article. - Herpes simplex virus type I glycoprotein L evades host antiviral innate immunity by abrogating the nuclear translocation of phosphorylated NF-κB sub-unit p65.
Li Z, Feng Z, Fang Z, Chen J, Chen W, Liang W, Chen Q. Li Z, et al. Front Microbiol. 2023 May 9;14:1178249. doi: 10.3389/fmicb.2023.1178249. eCollection 2023. Front Microbiol. 2023. PMID: 37228366 Free PMC article.
References
- Andrews, D. M., A. A. Scalzo, W. M. Yokoyama, M. J. Smyth, and M. A. Degli-Esposti. 2003. Functional interactions between dendritic cells and NK cells during viral infection. Nat. Immunol. 4:175-181. - PubMed
- Asselin-Paturel, C., A. Boonstra, M. Dalod, I. Durand, N. Yessaad, C. Dezutter-Dambuyant, A. Vicari, A. O'Garra, C. Biron, F. Briere, and G. Trinchieri. 2001. Mouse type I IFN-producing cells are immature APCs with plasmacytoid morphology. Nat. Immunol. 2:1144-1150. - PubMed
- Bjorck, P. 2001. Isolation and characterization of plasmacytoid dendritic cells from Flt3 ligand and granulocyte-macrophage colony-stimulating factor-treated mice. Blood 98:3520-3526. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Medical