miRNA profiling of primary lamb testicle cells infected with lumpy skin disease virus
Sakshi Pandita,
Assim Verma,
Himanshu Kamboj
et al.
Abstract:In this study, miRNA pro ling of cells infected with lumpy skin disease virus (LSDV) was conducted for the rst time. As compared to the mock-infected cells, LSDV-infected primary lamb testicle (LT) cells revealed dysregulation of 64, 85, and 85 miRNAs at 12 hours post-infection (hpi), 48 hpi and 72 hpi, respectively. While some of these miRNAs were found to be speci cally dysregulated at a particular time point following LSDV infection, others were commonly dysregulated across all three time points. The analys… Show more
“…Among the numerous dysregulated miRNAs that were identified in LSDV-infected primary lamb testicle cells [ 27 ], at least five more novel miRNAs were predicted for target sites on genes associated with regulation of interferon induction (Supplementary Figure S1).…”
Section: Resultsmentioning
confidence: 99%
“…In addition to regulating various physiological, metabolic, and developmental processes, miRNAs have been shown to regulate virus replication and immune responses [ 50 , 51 ]. We selected virus-associated dysregulated miRNAs [ 27 , 52 , 53 ] and confirmed the downregulation of circulating miR-29a levels in LSDV-infected cattle. miR-29a expression in NK cells, CD4 + T cells, and CD8 + T cells from Mycobacterium tuberculosis -infected mice was previously shown to suppress IFN-γ-production [ 52 , 54 ].…”
Section: Discussionmentioning
confidence: 99%
“…Taken together, it was concluded that miR-29a expression is indirectly involved in antagonizing the IFN-γ response in LSDV-infected cattle, and that reduced miR-29a expression may serve as a novel biomarker during the acute phase of LSDV infection. Among the hundreds of dysregulated miRNAs that have been identified in cells infected with LSDV [ 27 ], at least five more novel miRNAs were predicted to have target sites in genes that are associated with interferon regulation. Their further validation, along with elucidating their role in LSDV infection, is essential and needs further investigation.…”
Section: Discussionmentioning
confidence: 99%
“…miRNAs have also emerged as crucial regulators of the innate and adaptive immune responses [ 22–26 ]. We recently conducted miRNA profiling of LSDV-infected primary lamb testicular cells at various time points post-infection and identified several dysregulated miRNAs [ 27 ]. However, the specific roles of miRNAs in LSDV replication and pathogenesis remain unclear.…”
Micro RNAs (miRNAs) have been implicated in the regulation of maturation, proliferation, differentiation, and activation of immune cells. In this study, we demonstrated that miR-29a antagonizes IFN-γ production at early times post-LSDV infection in cattle. miR-29a was predicted to target upstream IFN-γ regulators, and its inhibition resulted in enhanced IFN-γ production in sensitized peripheral blood mononuclear cells (PBMCs). Further, stimulation of PBMCs with LSDV antigen exhibited lower levels of miR-29a, concomitant with a potent cell-mediated immune response (CMI), characterized by an increase in LSDV-specific CD8+ T cell counts and enhanced levels of IFN-γ, which eventually facilitated virus clearance. In addition, a few immunocompromised cattle (developed secondary LSDV infection at ~ 6 months) that failed to mount a potent cell-mediated immune response, were shown to maintain higher miR-29a levels. Furthermore, as compared to the sensitized crossbred cattle, PBMCs from sensitized Rathi (a native Indian breed) animals exhibited lower levels of miR-29a along with an increase in CD8+ T cell counts and enhanced levels of IFN-γ. Finally, we analysed that a ≥ 60% decrease in miR-29a expression levels in the PBMCs of sensitized cattle correlated with a potent CMI response. In conclusion, miR-29a expression is involved in antagonizing the IFN-γ response in LSDV-infected cattle and may serve as a novel biomarker for the acute phase of LSDV infection, as well as predicting the functionality of T cells in sensitized cattle. In addition, Rathi cattle mount a more potent CMI response against LSDV than crossbred cattle.
“…Among the numerous dysregulated miRNAs that were identified in LSDV-infected primary lamb testicle cells [ 27 ], at least five more novel miRNAs were predicted for target sites on genes associated with regulation of interferon induction (Supplementary Figure S1).…”
Section: Resultsmentioning
confidence: 99%
“…In addition to regulating various physiological, metabolic, and developmental processes, miRNAs have been shown to regulate virus replication and immune responses [ 50 , 51 ]. We selected virus-associated dysregulated miRNAs [ 27 , 52 , 53 ] and confirmed the downregulation of circulating miR-29a levels in LSDV-infected cattle. miR-29a expression in NK cells, CD4 + T cells, and CD8 + T cells from Mycobacterium tuberculosis -infected mice was previously shown to suppress IFN-γ-production [ 52 , 54 ].…”
Section: Discussionmentioning
confidence: 99%
“…Taken together, it was concluded that miR-29a expression is indirectly involved in antagonizing the IFN-γ response in LSDV-infected cattle, and that reduced miR-29a expression may serve as a novel biomarker during the acute phase of LSDV infection. Among the hundreds of dysregulated miRNAs that have been identified in cells infected with LSDV [ 27 ], at least five more novel miRNAs were predicted to have target sites in genes that are associated with interferon regulation. Their further validation, along with elucidating their role in LSDV infection, is essential and needs further investigation.…”
Section: Discussionmentioning
confidence: 99%
“…miRNAs have also emerged as crucial regulators of the innate and adaptive immune responses [ 22–26 ]. We recently conducted miRNA profiling of LSDV-infected primary lamb testicular cells at various time points post-infection and identified several dysregulated miRNAs [ 27 ]. However, the specific roles of miRNAs in LSDV replication and pathogenesis remain unclear.…”
Micro RNAs (miRNAs) have been implicated in the regulation of maturation, proliferation, differentiation, and activation of immune cells. In this study, we demonstrated that miR-29a antagonizes IFN-γ production at early times post-LSDV infection in cattle. miR-29a was predicted to target upstream IFN-γ regulators, and its inhibition resulted in enhanced IFN-γ production in sensitized peripheral blood mononuclear cells (PBMCs). Further, stimulation of PBMCs with LSDV antigen exhibited lower levels of miR-29a, concomitant with a potent cell-mediated immune response (CMI), characterized by an increase in LSDV-specific CD8+ T cell counts and enhanced levels of IFN-γ, which eventually facilitated virus clearance. In addition, a few immunocompromised cattle (developed secondary LSDV infection at ~ 6 months) that failed to mount a potent cell-mediated immune response, were shown to maintain higher miR-29a levels. Furthermore, as compared to the sensitized crossbred cattle, PBMCs from sensitized Rathi (a native Indian breed) animals exhibited lower levels of miR-29a along with an increase in CD8+ T cell counts and enhanced levels of IFN-γ. Finally, we analysed that a ≥ 60% decrease in miR-29a expression levels in the PBMCs of sensitized cattle correlated with a potent CMI response. In conclusion, miR-29a expression is involved in antagonizing the IFN-γ response in LSDV-infected cattle and may serve as a novel biomarker for the acute phase of LSDV infection, as well as predicting the functionality of T cells in sensitized cattle. In addition, Rathi cattle mount a more potent CMI response against LSDV than crossbred cattle.
“…While smallpox has been eradicated, the monkeypox virus (MPXV) has recently become an international concern for human health 1 . Likewise, the lumpy skin disease virus (LSDV) has emerged as an important pathogen in cattle population across many Asian countries with regard to animal health 2–7 . The poxviruses are usually host‐specific 8 .…”
In this study, we demonstrated the antiviral efficacy of hesperetin against multiple poxviruses, including buffalopox virus (BPXV), vaccinia virus (VACV), and lumpy skin disease virus (LSDV). The time‐of‐addition and virus step‐specific assays indicated that hesperetin reduces the levels of viral DNA, mRNA, and proteins in the target cells. Further, by immunoprecipitation (IP) of the viral RNA from BPXV‐infected Vero cells and a cell‐free RNA‐IP assay, we demonstrated that hesperetin‐induced reduction in BPXV protein synthesis is also consistent with diminished interaction between eukaryotic translation initiation factor eIF4E and the 5′ cap of viral mRNA. Molecular docking and MD simulation studies were also consistent with the binding of hesperetin to the cap‐binding pocket of eIF4E, adopting a conformation similar to m7GTP binding. Furthermore, in a BPXV egg infection model, hesperetin was shown to suppress the development of pock lesions on the chorioallantoic membrane and associated mortality in the chicken embryos. Most importantly, long‐term culture of BPXV in the presence of hesperetin did not induce the generation of drug‐resistant viral mutants. In conclusion, we, for the first time, demonstrated the antiviral activity of hesperetin against multiple poxviruses, besides providing some insights into its potential mechanisms of action.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.