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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-λ inhibits DENV-2 replication in a dose-dependent manner. C33-A cells were treated with increasing concentrations of IFN-α (a), IFN-λ1 (b) or IFN-λ2 (c) and then infected by DENV-2 (MOI = 0.1). Forty-eight hours post-infection, the viral titer was obtained by plaque assay. The doses used were 10–40 ng/mL for IFNλ1 or IFN-λ2 and 100–400 UI/ml for IFN-α. 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 = 8.705 ng/ml, IFN-λ2 = 8.24 ng/ml, IFN-α = 122–138 IU/ml. The points are representative of two independent experiments performed in triplicate. Experiments were fit to the Hill equation, using nonlinear regression to estimate the IC50 of IFN-λ1, IFN-λ2 or IFN-α (concentration of IFN that inhibited DENV-PFU production by 50 %, compared with parallel IFN free infected cultures). These IC50 values were considered for subsequent experimental procedures
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Fig5: IFN-λ inhibits DENV-2 replication in a dose-dependent manner. C33-A cells were treated with increasing concentrations of IFN-α (a), IFN-λ1 (b) or IFN-λ2 (c) and then infected by DENV-2 (MOI = 0.1). Forty-eight hours post-infection, the viral titer was obtained by plaque assay. The doses used were 10–40 ng/mL for IFNλ1 or IFN-λ2 and 100–400 UI/ml for IFN-α. 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 = 8.705 ng/ml, IFN-λ2 = 8.24 ng/ml, IFN-α = 122–138 IU/ml. The points are representative of two independent experiments performed in triplicate. Experiments were fit to the Hill equation, using nonlinear regression to estimate the IC50 of IFN-λ1, IFN-λ2 or IFN-α (concentration of IFN that inhibited DENV-PFU production by 50 %, compared with parallel IFN free infected cultures). These IC50 values were considered for subsequent experimental procedures

Mentions: To explore the effect of IFN-λ1 and IFN-λ2 on the DENV-2 infection, a replication inhibition curve was determined. C33-A cells were pretreated with increasing concentrations of IFN-λ1, IFN-λ2 (0, 10, 20, 30 and 40 ng/ml) or IFN-α (0, 100, 200, 300 and 400 IU/ml) prior to infection with DENV-2 (MOI = 0.1). Forty-eight hours after infection, the supernatants from each condition were quantified by lytic plaque-forming assays (Fig. 5).Fig. 5


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-λ inhibits DENV-2 replication in a dose-dependent manner. C33-A cells were treated with increasing concentrations of IFN-α (a), IFN-λ1 (b) or IFN-λ2 (c) and then infected by DENV-2 (MOI = 0.1). Forty-eight hours post-infection, the viral titer was obtained by plaque assay. The doses used were 10–40 ng/mL for IFNλ1 or IFN-λ2 and 100–400 UI/ml for IFN-α. 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 = 8.705 ng/ml, IFN-λ2 = 8.24 ng/ml, IFN-α = 122–138 IU/ml. The points are representative of two independent experiments performed in triplicate. Experiments were fit to the Hill equation, using nonlinear regression to estimate the IC50 of IFN-λ1, IFN-λ2 or IFN-α (concentration of IFN that inhibited DENV-PFU production by 50 %, compared with parallel IFN free infected cultures). These IC50 values were considered for subsequent experimental procedures
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig5: IFN-λ inhibits DENV-2 replication in a dose-dependent manner. C33-A cells were treated with increasing concentrations of IFN-α (a), IFN-λ1 (b) or IFN-λ2 (c) and then infected by DENV-2 (MOI = 0.1). Forty-eight hours post-infection, the viral titer was obtained by plaque assay. The doses used were 10–40 ng/mL for IFNλ1 or IFN-λ2 and 100–400 UI/ml for IFN-α. 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 = 8.705 ng/ml, IFN-λ2 = 8.24 ng/ml, IFN-α = 122–138 IU/ml. The points are representative of two independent experiments performed in triplicate. Experiments were fit to the Hill equation, using nonlinear regression to estimate the IC50 of IFN-λ1, IFN-λ2 or IFN-α (concentration of IFN that inhibited DENV-PFU production by 50 %, compared with parallel IFN free infected cultures). These IC50 values were considered for subsequent experimental procedures
Mentions: To explore the effect of IFN-λ1 and IFN-λ2 on the DENV-2 infection, a replication inhibition curve was determined. C33-A cells were pretreated with increasing concentrations of IFN-λ1, IFN-λ2 (0, 10, 20, 30 and 40 ng/ml) or IFN-α (0, 100, 200, 300 and 400 IU/ml) prior to infection with DENV-2 (MOI = 0.1). Forty-eight hours after infection, the supernatants from each condition were quantified by lytic plaque-forming assays (Fig. 5).Fig. 5

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