Limits...
Rabies virus infection induces type I interferon production in an IPS-1 dependent manner while dendritic cell activation relies on IFNAR signaling.

Faul EJ, Wanjalla CN, Suthar MS, Gale M, Wirblich C, Schnell MJ - PLoS Pathog. (2010)

Bottom Line: However, IPS-1 is essential for both BMDC activation and IFN production.In the absence of either receptor, there is a significant decrease in BMDC activation at 12h post infection.However, only RIG-I-/- cells exhibit a delay in type I IFN production.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania, United States of America.

ABSTRACT
As with many viruses, rabies virus (RABV) infection induces type I interferon (IFN) production within the infected host cells. However, RABV has evolved mechanisms by which to inhibit IFN production in order to sustain infection. Here we show that RABV infection of dendritic cells (DC) induces potent type I IFN production and DC activation. Although DCs are infected by RABV, the viral replication is highly suppressed in DCs, rendering the infection non-productive. We exploited this finding in bone marrow derived DCs (BMDC) in order to differentiate which pattern recognition receptor(s) (PRR) is responsible for inducing type I IFN following infection with RABV. Our results indicate that BMDC activation and type I IFN production following a RABV infection is independent of TLR signaling. However, IPS-1 is essential for both BMDC activation and IFN production. Interestingly, we see that the BMDC activation is primarily due to signaling through the IFNAR and only marginally induced by the initial infection. To further identify the receptor recognizing RABV infection, we next analyzed BMDC from Mda-5-/- and RIG-I-/- mice. In the absence of either receptor, there is a significant decrease in BMDC activation at 12h post infection. However, only RIG-I-/- cells exhibit a delay in type I IFN production. In order to determine the role that IPS-1 plays in vivo, we infected mice with pathogenic RABV. We see that IPS-1-/- mice are more susceptible to infection than IPS-1+/+ mice and have a significantly increased incident of limb paralysis.

Show MeSH

Related in: MedlinePlus

RABV infection of APCs, but not fibroblasts, induces type I IFN production.BSR, NA, Raw264.7, or JAWSII cells were infected with RABV (A) or UV-deactivated RABV (B) and the supernatant from the infected cells was UV-deactivated. Reporter cells were pre-treated with the UV-deactivated supernatant (diluted 1∶10) for 24h and then infected with VSV-GFP for 5h. Fluorescence indicates viral replication, and thus, lack of type I IFN. Photos are representative of two independent experiments.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2908622&req=5

ppat-1001016-g001: RABV infection of APCs, but not fibroblasts, induces type I IFN production.BSR, NA, Raw264.7, or JAWSII cells were infected with RABV (A) or UV-deactivated RABV (B) and the supernatant from the infected cells was UV-deactivated. Reporter cells were pre-treated with the UV-deactivated supernatant (diluted 1∶10) for 24h and then infected with VSV-GFP for 5h. Fluorescence indicates viral replication, and thus, lack of type I IFN. Photos are representative of two independent experiments.

Mentions: In order to check for type I IFN production, we first infected a variety of cell types including fibroblasts (BSR), neuronal cells (NA), macrophages (Raw264.7) and DCs (JAWSII) with a RABV vaccine strain-based vector, SPBN. Following infection with RABV, cell supernatants were collected and subsequently UV-treated in order to deactivate any infectious virus but retain secreted cellular proteins, such as type I IFN. We then transferred the supernatants to reporter cells, which are sensitive to IFN. Twenty-four hours after supernatant transfer, reporter cells were infected with recombinant vesicular stomatitis virus expressing GFP (VSV-GFP, [31]) for 5–8h. VSV replication is highly sensitive to type I IFN [32], and thus, in the presence of type I IFN, the replication of VSV is suppressed [4]. Following infection with RABV, macrophages as well as DCs, but not fibroblasts or neuronal cells, produce type I IFN that inhibits VSV-GFP replication, as indicated by the lack of GFP expression (Figure 1A). Of note, when BSR, NA, Raw264.7, or JAWSII cells are originally treated with UV-deactivatecd RABV, the supernatants from these cells are unable to block VSV replication (Figure 1B); therefore, IFN is secreted only after RABV replication.


Rabies virus infection induces type I interferon production in an IPS-1 dependent manner while dendritic cell activation relies on IFNAR signaling.

Faul EJ, Wanjalla CN, Suthar MS, Gale M, Wirblich C, Schnell MJ - PLoS Pathog. (2010)

RABV infection of APCs, but not fibroblasts, induces type I IFN production.BSR, NA, Raw264.7, or JAWSII cells were infected with RABV (A) or UV-deactivated RABV (B) and the supernatant from the infected cells was UV-deactivated. Reporter cells were pre-treated with the UV-deactivated supernatant (diluted 1∶10) for 24h and then infected with VSV-GFP for 5h. Fluorescence indicates viral replication, and thus, lack of type I IFN. Photos are representative of two independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2908622&req=5

ppat-1001016-g001: RABV infection of APCs, but not fibroblasts, induces type I IFN production.BSR, NA, Raw264.7, or JAWSII cells were infected with RABV (A) or UV-deactivated RABV (B) and the supernatant from the infected cells was UV-deactivated. Reporter cells were pre-treated with the UV-deactivated supernatant (diluted 1∶10) for 24h and then infected with VSV-GFP for 5h. Fluorescence indicates viral replication, and thus, lack of type I IFN. Photos are representative of two independent experiments.
Mentions: In order to check for type I IFN production, we first infected a variety of cell types including fibroblasts (BSR), neuronal cells (NA), macrophages (Raw264.7) and DCs (JAWSII) with a RABV vaccine strain-based vector, SPBN. Following infection with RABV, cell supernatants were collected and subsequently UV-treated in order to deactivate any infectious virus but retain secreted cellular proteins, such as type I IFN. We then transferred the supernatants to reporter cells, which are sensitive to IFN. Twenty-four hours after supernatant transfer, reporter cells were infected with recombinant vesicular stomatitis virus expressing GFP (VSV-GFP, [31]) for 5–8h. VSV replication is highly sensitive to type I IFN [32], and thus, in the presence of type I IFN, the replication of VSV is suppressed [4]. Following infection with RABV, macrophages as well as DCs, but not fibroblasts or neuronal cells, produce type I IFN that inhibits VSV-GFP replication, as indicated by the lack of GFP expression (Figure 1A). Of note, when BSR, NA, Raw264.7, or JAWSII cells are originally treated with UV-deactivatecd RABV, the supernatants from these cells are unable to block VSV replication (Figure 1B); therefore, IFN is secreted only after RABV replication.

Bottom Line: However, IPS-1 is essential for both BMDC activation and IFN production.In the absence of either receptor, there is a significant decrease in BMDC activation at 12h post infection.However, only RIG-I-/- cells exhibit a delay in type I IFN production.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania, United States of America.

ABSTRACT
As with many viruses, rabies virus (RABV) infection induces type I interferon (IFN) production within the infected host cells. However, RABV has evolved mechanisms by which to inhibit IFN production in order to sustain infection. Here we show that RABV infection of dendritic cells (DC) induces potent type I IFN production and DC activation. Although DCs are infected by RABV, the viral replication is highly suppressed in DCs, rendering the infection non-productive. We exploited this finding in bone marrow derived DCs (BMDC) in order to differentiate which pattern recognition receptor(s) (PRR) is responsible for inducing type I IFN following infection with RABV. Our results indicate that BMDC activation and type I IFN production following a RABV infection is independent of TLR signaling. However, IPS-1 is essential for both BMDC activation and IFN production. Interestingly, we see that the BMDC activation is primarily due to signaling through the IFNAR and only marginally induced by the initial infection. To further identify the receptor recognizing RABV infection, we next analyzed BMDC from Mda-5-/- and RIG-I-/- mice. In the absence of either receptor, there is a significant decrease in BMDC activation at 12h post infection. However, only RIG-I-/- cells exhibit a delay in type I IFN production. In order to determine the role that IPS-1 plays in vivo, we infected mice with pathogenic RABV. We see that IPS-1-/- mice are more susceptible to infection than IPS-1+/+ mice and have a significantly increased incident of limb paralysis.

Show MeSH
Related in: MedlinePlus