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Interferon lambda inhibits dengue virus replication in epithelial cells.

Palma-Ocampo HK, Flores-Alonso JC, Vallejo-Ruiz V, Reyes-Leyva J, Flores-Mendoza L, Herrera-Camacho I, Rosas-Murrieta NH, Santos-López G - Virol. J. (2015)

Bottom Line: We found increased (~1.8 times) serological IFN-λ in dengue fever patients compared to healthy blood donors.The reduction of viral titer corresponded with increased ISG mRNA levels (MX1 and OAS1), with the highest inhibition occurring at ISG's peak expression.Mechanisms for the cellular and organismal interplay between DENV and IFN- λ need to be further studied as they could provide insights into strategies to treat this disease.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Biología Molecular y Virología, Centro de Investigación Biomédica de Oriente, Instituto Mexicano del Seguro Social, Metepec, Puebla, México. hk.palma@gmail.com.

ABSTRACT

Background: In viral disease, infection is controlled at the cellular level by type I interferon (IFN-I), but dengue virus (DENV) has the ability to inhibit this response. Type III interferon, also known as lambda IFN (IFN-III or IFN-λ), is a complementary pathway to the antiviral response by IFN-I. This work analyzed the IFN-λ (IFN-III) mediated antiviral response against DENV serotype 2 (DENV-2) infection.

Methods: Dengue fever patients were sampled to determine their IFN-λ levels by ELISA. To study the IFN-λ response during DENV infection we selected the epithelial cell line C33-A, and we demonstrated that it is permissive to DENV-2 infection. The effect of IFN-λ on virus replication was determined in these cells, in parallel to the expression of IFN-stimulated genes (ISGs), and Suppressor of Cytokine Signaling (SOCS), genes measured by RT-qPCR.

Results: We found increased (~1.8 times) serological IFN-λ in dengue fever patients compared to healthy blood donors. IFN-λ inhibited DENV-2 replication in a dose-dependent manner in vitro. The reduction of viral titer corresponded with increased ISG mRNA levels (MX1 and OAS1), with the highest inhibition occurring at ISG's peak expression. Presence of IFN-negative regulators, SOCS1 and SOCS3, during DENV-2 infection was associated with reduced IFN-λ1 expression.

Conclusions: Evidence described here suggests that IFN-λ is a good candidate inhibitor of viral replication in dengue infection. Mechanisms for the cellular and organismal interplay between DENV and IFN- λ need to be further studied as they could provide insights into strategies to treat this disease. Furthermore, we report a novel epithelial model to study dengue infection in vitro.

No MeSH data available.


Related in: MedlinePlus

IFN-λ1 inhibits DENV-2 replication in cells lacking type I IFN. Vero cells were treated with increasing concentrations of IFN-α (a) or IFN-λ1 (b) or IFN-λ1 plus IFN-α (c), and then infected by DENV-2 (MOI = 0.1). The doses used were 10–40 ng/ml of IFN-λ1 or 100–400 UI/ml of IFN-α. Forty-eight hours post-infection, the viral titer was obtained by plaque assay. Plots show the percentage inhibition of the viral titer in each group compared to untreated cells. Dotted lines represent the IC50 values corresponding to IFN-λ1 = 37.15 ng/ml, IFN-λ1 in presence of IFN-α = 14.12 ng/ml, the value of IC50 for IFN-α was not calculated because 50 % inhibition was not reached under the conditions of experiment. Error bars represent standard deviations of means from two independent experiment *p < 0.01, **p < 0.001
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Fig7: IFN-λ1 inhibits DENV-2 replication in cells lacking type I IFN. Vero cells were treated with increasing concentrations of IFN-α (a) or IFN-λ1 (b) or IFN-λ1 plus IFN-α (c), and then infected by DENV-2 (MOI = 0.1). The doses used were 10–40 ng/ml of IFN-λ1 or 100–400 UI/ml of IFN-α. Forty-eight hours post-infection, the viral titer was obtained by plaque assay. Plots show the percentage inhibition of the viral titer in each group compared to untreated cells. Dotted lines represent the IC50 values corresponding to IFN-λ1 = 37.15 ng/ml, IFN-λ1 in presence of IFN-α = 14.12 ng/ml, the value of IC50 for IFN-α was not calculated because 50 % inhibition was not reached under the conditions of experiment. Error bars represent standard deviations of means from two independent experiment *p < 0.01, **p < 0.001

Mentions: To determine whether IFN-λ1 is enough to inhibit replication of DENV-2, we tested its ability to inhibit viral replication in the absence of INF-α. Inhibition experiments were similar to those described in Fig. 5, but they were performed on Vero cells that do not produce IFN-α, yet respond to it if supplied externally [21]. Vero cells were pre-incubated for 6 h with increasing concentrations of IFN-α, IFN-λ1 or IFN-λ1 plus IFN-α and then infected with DENV-2. By itself, IFN-λ1 treatment decreased the viral progeny 50 % at 37.15 ng/ml (IC50) (Fig. 7b). Treatment with IFN-λ1 plus IFN-α had a lower IC50 of 14.12 ng/ml calculated for IFN-λ1 (Fig. 7c); while for IFN-α treatment alone, IC50 couldn’t be calculated because the 50 % inhibition was not reached under the experimental conditions (Fig. 7a). This experiment suggests that IFN-λ1 is able to inhibit DENV-2 infection, but an effective response likely involves both types of interferons and a possible synergism between them.Fig. 7


Interferon lambda inhibits dengue virus replication in epithelial cells.

Palma-Ocampo HK, Flores-Alonso JC, Vallejo-Ruiz V, Reyes-Leyva J, Flores-Mendoza L, Herrera-Camacho I, Rosas-Murrieta NH, Santos-López G - Virol. J. (2015)

IFN-λ1 inhibits DENV-2 replication in cells lacking type I IFN. Vero cells were treated with increasing concentrations of IFN-α (a) or IFN-λ1 (b) or IFN-λ1 plus IFN-α (c), and then infected by DENV-2 (MOI = 0.1). The doses used were 10–40 ng/ml of IFN-λ1 or 100–400 UI/ml of IFN-α. Forty-eight hours post-infection, the viral titer was obtained by plaque assay. Plots show the percentage inhibition of the viral titer in each group compared to untreated cells. Dotted lines represent the IC50 values corresponding to IFN-λ1 = 37.15 ng/ml, IFN-λ1 in presence of IFN-α = 14.12 ng/ml, the value of IC50 for IFN-α was not calculated because 50 % inhibition was not reached under the conditions of experiment. Error bars represent standard deviations of means from two independent experiment *p < 0.01, **p < 0.001
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4584467&req=5

Fig7: IFN-λ1 inhibits DENV-2 replication in cells lacking type I IFN. Vero cells were treated with increasing concentrations of IFN-α (a) or IFN-λ1 (b) or IFN-λ1 plus IFN-α (c), and then infected by DENV-2 (MOI = 0.1). The doses used were 10–40 ng/ml of IFN-λ1 or 100–400 UI/ml of IFN-α. Forty-eight hours post-infection, the viral titer was obtained by plaque assay. Plots show the percentage inhibition of the viral titer in each group compared to untreated cells. Dotted lines represent the IC50 values corresponding to IFN-λ1 = 37.15 ng/ml, IFN-λ1 in presence of IFN-α = 14.12 ng/ml, the value of IC50 for IFN-α was not calculated because 50 % inhibition was not reached under the conditions of experiment. Error bars represent standard deviations of means from two independent experiment *p < 0.01, **p < 0.001
Mentions: To determine whether IFN-λ1 is enough to inhibit replication of DENV-2, we tested its ability to inhibit viral replication in the absence of INF-α. Inhibition experiments were similar to those described in Fig. 5, but they were performed on Vero cells that do not produce IFN-α, yet respond to it if supplied externally [21]. Vero cells were pre-incubated for 6 h with increasing concentrations of IFN-α, IFN-λ1 or IFN-λ1 plus IFN-α and then infected with DENV-2. By itself, IFN-λ1 treatment decreased the viral progeny 50 % at 37.15 ng/ml (IC50) (Fig. 7b). Treatment with IFN-λ1 plus IFN-α had a lower IC50 of 14.12 ng/ml calculated for IFN-λ1 (Fig. 7c); while for IFN-α treatment alone, IC50 couldn’t be calculated because the 50 % inhibition was not reached under the experimental conditions (Fig. 7a). This experiment suggests that IFN-λ1 is able to inhibit DENV-2 infection, but an effective response likely involves both types of interferons and a possible synergism between them.Fig. 7

Bottom Line: We found increased (~1.8 times) serological IFN-λ in dengue fever patients compared to healthy blood donors.The reduction of viral titer corresponded with increased ISG mRNA levels (MX1 and OAS1), with the highest inhibition occurring at ISG's peak expression.Mechanisms for the cellular and organismal interplay between DENV and IFN- λ need to be further studied as they could provide insights into strategies to treat this disease.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Biología Molecular y Virología, Centro de Investigación Biomédica de Oriente, Instituto Mexicano del Seguro Social, Metepec, Puebla, México. hk.palma@gmail.com.

ABSTRACT

Background: In viral disease, infection is controlled at the cellular level by type I interferon (IFN-I), but dengue virus (DENV) has the ability to inhibit this response. Type III interferon, also known as lambda IFN (IFN-III or IFN-λ), is a complementary pathway to the antiviral response by IFN-I. This work analyzed the IFN-λ (IFN-III) mediated antiviral response against DENV serotype 2 (DENV-2) infection.

Methods: Dengue fever patients were sampled to determine their IFN-λ levels by ELISA. To study the IFN-λ response during DENV infection we selected the epithelial cell line C33-A, and we demonstrated that it is permissive to DENV-2 infection. The effect of IFN-λ on virus replication was determined in these cells, in parallel to the expression of IFN-stimulated genes (ISGs), and Suppressor of Cytokine Signaling (SOCS), genes measured by RT-qPCR.

Results: We found increased (~1.8 times) serological IFN-λ in dengue fever patients compared to healthy blood donors. IFN-λ inhibited DENV-2 replication in a dose-dependent manner in vitro. The reduction of viral titer corresponded with increased ISG mRNA levels (MX1 and OAS1), with the highest inhibition occurring at ISG's peak expression. Presence of IFN-negative regulators, SOCS1 and SOCS3, during DENV-2 infection was associated with reduced IFN-λ1 expression.

Conclusions: Evidence described here suggests that IFN-λ is a good candidate inhibitor of viral replication in dengue infection. Mechanisms for the cellular and organismal interplay between DENV and IFN- λ need to be further studied as they could provide insights into strategies to treat this disease. Furthermore, we report a novel epithelial model to study dengue infection in vitro.

No MeSH data available.


Related in: MedlinePlus