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Suppressed expression of miR-378 targeting gzmb in NK cells is required to control dengue virus infection

View Article: PubMed Central - PubMed

ABSTRACT

Dengue virus (DENV) remains a major public health threat because no vaccine or drugs are available for the prevention and treatment of DENV infection, and the immunopathogenesis mechanisms of DENV infection are not fully understood. Cytotoxic molecules, such as granzyme B (GrzB), may be necessary to control viral infections. However, the exact role of GrzB during DENV infection and the mechanisms regulating GrzB expression during DENV infection are not clear. This study found that miR-27a*, miR-30e, and miR-378 were down-regulated in DENV-infected patients, and DENV infection in humans induced a significant up-regulation of GrzB in natural killer (NK) cells and CD8+ T cells. Further investigation indicated that NK cells, but not CD8+ T cells, were the major sources of GrzB, and miR-378, but not miR-27a* or miR-30e, suppressed GrzB expression in NK cells. Notably, we found that overexpression of miR-378 using a miR-378 agomir in DENV-infected mice inhibited GrzB expression and promoted DENV replication. These results suggest the critical importance of miR-378 in the regulation of GrzB expression and a protective role for GrzB in controlling DENV replication in vivo. Therefore, this study provides a new insight into the immunopathogenesis mechanism of DENV infection and a biological basis for the development of new therapeutic strategies to control DENV infection.

No MeSH data available.


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miR-27a*, miR-378, and miR-30e directly target perforin and/or GrzB, which are significantly down-regulated in DENV patients. (a) Human perforin and/or GrzB are putative targets of miR-27a*, miR-30e, and/or miR-378, as predicted by miRanda and TargetScan. Numbers indicate the position of nucleotides in the 3′-UTR that are targeted by miRNAs. (b) Pooled data show the expression levels of miR-27a*, miR-30e, and miR-378 in PBMCs of DENV-infected patients (DENV patients), which is much lower than expression in PBMCs of Healthy Ctrls. Freshly isolated PBMCs were obtained from 10 DENV-infected patients and 10 Healthy Ctrls, and total RNA was extracted for analyses of miRNA expression using qPCR analysis. Data are representative of three independent experiments (mean ± SD; independent samples t-test, **p < 0.01, ***p < 0.001).
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fig1: miR-27a*, miR-378, and miR-30e directly target perforin and/or GrzB, which are significantly down-regulated in DENV patients. (a) Human perforin and/or GrzB are putative targets of miR-27a*, miR-30e, and/or miR-378, as predicted by miRanda and TargetScan. Numbers indicate the position of nucleotides in the 3′-UTR that are targeted by miRNAs. (b) Pooled data show the expression levels of miR-27a*, miR-30e, and miR-378 in PBMCs of DENV-infected patients (DENV patients), which is much lower than expression in PBMCs of Healthy Ctrls. Freshly isolated PBMCs were obtained from 10 DENV-infected patients and 10 Healthy Ctrls, and total RNA was extracted for analyses of miRNA expression using qPCR analysis. Data are representative of three independent experiments (mean ± SD; independent samples t-test, **p < 0.01, ***p < 0.001).

Mentions: We used miRanda and TargetScan software to predict the sequence of miRNAs that potentially bind the 3′-UTR regions of perforin and GrzB mRNA to determine whether miRNAs regulate the expression of these human cytotoxic molecules. MiR-27a*, miR-30e, and miR-378 most potently targeted perforin and GrzB (Figure 1a), which suggests that miR-27a*, miR-30e, and miR-378 are the primary miRNAs that regulate perforin and GrzB expression. Previous studies suggested a role of miR-27a*, miR-30e, and miR-378 in the regulation of perforin and GrzB.22,23 Therefore, we chose miR-27a*, miR-30e, and miR-378 as targets to determine the relationship between miRNA expression and perforin and GrzB production.


Suppressed expression of miR-378 targeting gzmb in NK cells is required to control dengue virus infection
miR-27a*, miR-378, and miR-30e directly target perforin and/or GrzB, which are significantly down-regulated in DENV patients. (a) Human perforin and/or GrzB are putative targets of miR-27a*, miR-30e, and/or miR-378, as predicted by miRanda and TargetScan. Numbers indicate the position of nucleotides in the 3′-UTR that are targeted by miRNAs. (b) Pooled data show the expression levels of miR-27a*, miR-30e, and miR-378 in PBMCs of DENV-infected patients (DENV patients), which is much lower than expression in PBMCs of Healthy Ctrls. Freshly isolated PBMCs were obtained from 10 DENV-infected patients and 10 Healthy Ctrls, and total RNA was extracted for analyses of miRNA expression using qPCR analysis. Data are representative of three independent experiments (mean ± SD; independent samples t-test, **p < 0.01, ***p < 0.001).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5037283&req=5

fig1: miR-27a*, miR-378, and miR-30e directly target perforin and/or GrzB, which are significantly down-regulated in DENV patients. (a) Human perforin and/or GrzB are putative targets of miR-27a*, miR-30e, and/or miR-378, as predicted by miRanda and TargetScan. Numbers indicate the position of nucleotides in the 3′-UTR that are targeted by miRNAs. (b) Pooled data show the expression levels of miR-27a*, miR-30e, and miR-378 in PBMCs of DENV-infected patients (DENV patients), which is much lower than expression in PBMCs of Healthy Ctrls. Freshly isolated PBMCs were obtained from 10 DENV-infected patients and 10 Healthy Ctrls, and total RNA was extracted for analyses of miRNA expression using qPCR analysis. Data are representative of three independent experiments (mean ± SD; independent samples t-test, **p < 0.01, ***p < 0.001).
Mentions: We used miRanda and TargetScan software to predict the sequence of miRNAs that potentially bind the 3′-UTR regions of perforin and GrzB mRNA to determine whether miRNAs regulate the expression of these human cytotoxic molecules. MiR-27a*, miR-30e, and miR-378 most potently targeted perforin and GrzB (Figure 1a), which suggests that miR-27a*, miR-30e, and miR-378 are the primary miRNAs that regulate perforin and GrzB expression. Previous studies suggested a role of miR-27a*, miR-30e, and miR-378 in the regulation of perforin and GrzB.22,23 Therefore, we chose miR-27a*, miR-30e, and miR-378 as targets to determine the relationship between miRNA expression and perforin and GrzB production.

View Article: PubMed Central - PubMed

ABSTRACT

Dengue virus (DENV) remains a major public health threat because no vaccine or drugs are available for the prevention and treatment of DENV infection, and the immunopathogenesis mechanisms of DENV infection are not fully understood. Cytotoxic molecules, such as granzyme B (GrzB), may be necessary to control viral infections. However, the exact role of GrzB during DENV infection and the mechanisms regulating GrzB expression during DENV infection are not clear. This study found that miR-27a*, miR-30e, and miR-378 were down-regulated in DENV-infected patients, and DENV infection in humans induced a significant up-regulation of GrzB in natural killer (NK) cells and CD8+ T cells. Further investigation indicated that NK cells, but not CD8+ T cells, were the major sources of GrzB, and miR-378, but not miR-27a* or miR-30e, suppressed GrzB expression in NK cells. Notably, we found that overexpression of miR-378 using a miR-378 agomir in DENV-infected mice inhibited GrzB expression and promoted DENV replication. These results suggest the critical importance of miR-378 in the regulation of GrzB expression and a protective role for GrzB in controlling DENV replication in vivo. Therefore, this study provides a new insight into the immunopathogenesis mechanism of DENV infection and a biological basis for the development of new therapeutic strategies to control DENV infection.

No MeSH data available.


Related in: MedlinePlus