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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

Transcription levels of IFN-stimulated genes during DENV-2 infection. Expression of OAS1 (a), MX1 (b), IFN-β (c), IFN-λ1 (d), SOCS1 (e), and SOCS3 genes (f) in cells infected with DENV-2 (MOI = 0.1) or treated with 10 ng/ml of IFN-λ1 or treated with IFN-λ1 before DENV-2 infection, were determined by RT-qPCR at 0, 6, 12, 18, 24, and 48 h post-infection or post-treatment. Results are presented as expression relative to untreated cells, which have a value of 1. mRNA levels were normalized to the HPRT gene. Data points represent the mean of three replicates and error bars indicate standard deviation
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Fig8: Transcription levels of IFN-stimulated genes during DENV-2 infection. Expression of OAS1 (a), MX1 (b), IFN-β (c), IFN-λ1 (d), SOCS1 (e), and SOCS3 genes (f) in cells infected with DENV-2 (MOI = 0.1) or treated with 10 ng/ml of IFN-λ1 or treated with IFN-λ1 before DENV-2 infection, were determined by RT-qPCR at 0, 6, 12, 18, 24, and 48 h post-infection or post-treatment. Results are presented as expression relative to untreated cells, which have a value of 1. mRNA levels were normalized to the HPRT gene. Data points represent the mean of three replicates and error bars indicate standard deviation

Mentions: Figure 8a shows that stimulation of C33-A cells with IFN-λ1 slightly increases the transcription of OAS1 to similar levels between 6 and 48 h post-treatment (pt). DENV-2 infection significantly increases OAS1 mRNA but at earlier times, peaking at 6 h post-infection. However, the most striking effect was present with IFN-λ1 pre-treatment followed by DENV-2 infection, that showed a pronounced increase in OAS1 expression from 6 to 18 h post-infection, at which time it peaked and then underwent a slow steady decrease, remaining high at 24 and 48 h. This result suggests that IFN-λ1 decreases DENV-2 replication in part by viral mRNA degradation via RNase L. Moreover, stimulation of cells with IFN-λ1 dramatically increases MX1 gene transcription, suggesting that the efficient activation of IFN-III signal transduction is useful in controlling DENV-2 infection. IFN-λ1 treatment induced a bimodal MX1 increase with a first peak at 6 h post-infection and a second peak nearly 100-fold higher than basal at 24 h (Fig. 8b). Bimodality probably is due as a result of a first wave of activation by IFN-β and IFN-λ1 and subsequently, the increase at 24 h could be by IFN-λ2, which has demonstrated late induction, as well as by the late wave of endogenous interferons [22]. MX1 mRNA was detected at similar levels in infected and non-infected cells at all the studied times, suggesting that DENV induces an impairment in IFN signal transduction; however, this experiment cannot discriminate between IFN-I or IFN-III specific pathways. In cells pre-treated with IFN-λ1 and then infected, MX1 mRNA increased between 18 and 48 h post-infection, although not as much as OAS1.Fig. 8


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)

Transcription levels of IFN-stimulated genes during DENV-2 infection. Expression of OAS1 (a), MX1 (b), IFN-β (c), IFN-λ1 (d), SOCS1 (e), and SOCS3 genes (f) in cells infected with DENV-2 (MOI = 0.1) or treated with 10 ng/ml of IFN-λ1 or treated with IFN-λ1 before DENV-2 infection, were determined by RT-qPCR at 0, 6, 12, 18, 24, and 48 h post-infection or post-treatment. Results are presented as expression relative to untreated cells, which have a value of 1. mRNA levels were normalized to the HPRT gene. Data points represent the mean of three replicates and error bars indicate standard deviation
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig8: Transcription levels of IFN-stimulated genes during DENV-2 infection. Expression of OAS1 (a), MX1 (b), IFN-β (c), IFN-λ1 (d), SOCS1 (e), and SOCS3 genes (f) in cells infected with DENV-2 (MOI = 0.1) or treated with 10 ng/ml of IFN-λ1 or treated with IFN-λ1 before DENV-2 infection, were determined by RT-qPCR at 0, 6, 12, 18, 24, and 48 h post-infection or post-treatment. Results are presented as expression relative to untreated cells, which have a value of 1. mRNA levels were normalized to the HPRT gene. Data points represent the mean of three replicates and error bars indicate standard deviation
Mentions: Figure 8a shows that stimulation of C33-A cells with IFN-λ1 slightly increases the transcription of OAS1 to similar levels between 6 and 48 h post-treatment (pt). DENV-2 infection significantly increases OAS1 mRNA but at earlier times, peaking at 6 h post-infection. However, the most striking effect was present with IFN-λ1 pre-treatment followed by DENV-2 infection, that showed a pronounced increase in OAS1 expression from 6 to 18 h post-infection, at which time it peaked and then underwent a slow steady decrease, remaining high at 24 and 48 h. This result suggests that IFN-λ1 decreases DENV-2 replication in part by viral mRNA degradation via RNase L. Moreover, stimulation of cells with IFN-λ1 dramatically increases MX1 gene transcription, suggesting that the efficient activation of IFN-III signal transduction is useful in controlling DENV-2 infection. IFN-λ1 treatment induced a bimodal MX1 increase with a first peak at 6 h post-infection and a second peak nearly 100-fold higher than basal at 24 h (Fig. 8b). Bimodality probably is due as a result of a first wave of activation by IFN-β and IFN-λ1 and subsequently, the increase at 24 h could be by IFN-λ2, which has demonstrated late induction, as well as by the late wave of endogenous interferons [22]. MX1 mRNA was detected at similar levels in infected and non-infected cells at all the studied times, suggesting that DENV induces an impairment in IFN signal transduction; however, this experiment cannot discriminate between IFN-I or IFN-III specific pathways. In cells pre-treated with IFN-λ1 and then infected, MX1 mRNA increased between 18 and 48 h post-infection, although not as much as OAS1.Fig. 8

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