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Proof-of-concept study: profile of circulating microRNAs in Bovine serum harvested during acute and persistent FMDV infection

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ABSTRACT

Background: Changes in the levels of circulating microRNAs (miRNAs) in the serum of humans and animals have been detected as a result of infection with a variety of viruses. However, to date, such a miRNA profiling study has not been conducted for foot-and-mouth disease virus (FMDV) infection.

Methods: The relative abundance of 169 miRNAs was measured in bovine serum collected at three different phases of FMDV infection in a proof-of-concept study using miRNA PCR array plates.

Results: Alterations in specific miRNA levels were detected in serum during acute, persistent, and convalescent phases of FMDV infection. Subclinical FMDV persistence produced a circulating miRNA profile distinct from cattle that had cleared infection. bta-miR-17-5p was highest expressed during acute infection, whereas bta-miR-31 was the highest during FMDV persistence. Interestingly, miR-1281was significantly down-regulated during both acute and persistent infection. Cattle that cleared infection resembled the baseline profile, adding support to applying serum miRNA profiling for identification of sub-clinically infected FMDV carriers. Significantly regulated miRNAs during acute or persistent infection were associated with cellular proliferation, apoptosis, modulation of the immune response, and lipid metabolism.

Conclusions: These findings suggest a role for non-coding regulatory RNAs in FMDV infection of cattle. Future studies will delineate the individual contributions of the reported miRNAs to FMDV replication, determine if this miRNA signature is applicable across all FMDV serotypes, and may facilitate development of novel diagnostic applications.

Electronic supplementary material: The online version of this article (doi:10.1186/s12985-017-0743-3) contains supplementary material, which is available to authorized users.

No MeSH data available.


Related in: MedlinePlus

Schematic overview of miRNA profiling study. a Serum was collected from three different groups of FMDV-infected cattle: acutely infected (viremic; 3–4 dpi), persistently infected (“FMDV carriers”; 35 dpi) and convalescent (“non-carriers”; 35 dpi), and were compared to uninfected controls. Each group comprised serum samples from three animals. b miRNAs were purified from individual serum samples. The purified miRNAs were reverse transcribed into complementary DNA (cDNA). The cDNA samples were then analyzed by RT-PCR on bovine miRNome miRNA array plates containing primers to 169 different bovine miRNAs. The results obtained indicated which miRNAs were left unchanged, up-regulated, or down-regulated in response to FMDV infection. c schematic of the organization of the miRNA PCR array plates utilized in this study
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Fig1: Schematic overview of miRNA profiling study. a Serum was collected from three different groups of FMDV-infected cattle: acutely infected (viremic; 3–4 dpi), persistently infected (“FMDV carriers”; 35 dpi) and convalescent (“non-carriers”; 35 dpi), and were compared to uninfected controls. Each group comprised serum samples from three animals. b miRNAs were purified from individual serum samples. The purified miRNAs were reverse transcribed into complementary DNA (cDNA). The cDNA samples were then analyzed by RT-PCR on bovine miRNome miRNA array plates containing primers to 169 different bovine miRNAs. The results obtained indicated which miRNAs were left unchanged, up-regulated, or down-regulated in response to FMDV infection. c schematic of the organization of the miRNA PCR array plates utilized in this study

Mentions: This current work comprises a proof-of-concept study in which the differential regulation of serum miRNAs was investigated in cattle infected with FMDV serotype A isolate A24 Cruzeiro. The bovine serum miRNA profile was characterized using quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) array plates designed to detect the relative abundance levels of 169 of the bovine miRNAs characterized in the past several years (Table 1, Fig. 1c). Serum from uninfected animals was compared to samples obtained through three distinct phases of FMDV infection: acute infection (peak viremia), persistent infection (subclinical persistence of FMDV in the upper respiratory tract), and convalescent phase, comprising animals that had successfully cleared the infection (Fig. 1a, b). Distinct miRNA signatures of up- and down-regulated miRNAs in circulation in response to FMDV infection were detected. The results obtained from this proof-of-concept study reinforce the application of this approach to future large scale testing and the investigation of miRNA signatures for other serotypes of FMDV.Table 1


Proof-of-concept study: profile of circulating microRNAs in Bovine serum harvested during acute and persistent FMDV infection
Schematic overview of miRNA profiling study. a Serum was collected from three different groups of FMDV-infected cattle: acutely infected (viremic; 3–4 dpi), persistently infected (“FMDV carriers”; 35 dpi) and convalescent (“non-carriers”; 35 dpi), and were compared to uninfected controls. Each group comprised serum samples from three animals. b miRNAs were purified from individual serum samples. The purified miRNAs were reverse transcribed into complementary DNA (cDNA). The cDNA samples were then analyzed by RT-PCR on bovine miRNome miRNA array plates containing primers to 169 different bovine miRNAs. The results obtained indicated which miRNAs were left unchanged, up-regulated, or down-regulated in response to FMDV infection. c schematic of the organization of the miRNA PCR array plates utilized in this study
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC5384155&req=5

Fig1: Schematic overview of miRNA profiling study. a Serum was collected from three different groups of FMDV-infected cattle: acutely infected (viremic; 3–4 dpi), persistently infected (“FMDV carriers”; 35 dpi) and convalescent (“non-carriers”; 35 dpi), and were compared to uninfected controls. Each group comprised serum samples from three animals. b miRNAs were purified from individual serum samples. The purified miRNAs were reverse transcribed into complementary DNA (cDNA). The cDNA samples were then analyzed by RT-PCR on bovine miRNome miRNA array plates containing primers to 169 different bovine miRNAs. The results obtained indicated which miRNAs were left unchanged, up-regulated, or down-regulated in response to FMDV infection. c schematic of the organization of the miRNA PCR array plates utilized in this study
Mentions: This current work comprises a proof-of-concept study in which the differential regulation of serum miRNAs was investigated in cattle infected with FMDV serotype A isolate A24 Cruzeiro. The bovine serum miRNA profile was characterized using quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) array plates designed to detect the relative abundance levels of 169 of the bovine miRNAs characterized in the past several years (Table 1, Fig. 1c). Serum from uninfected animals was compared to samples obtained through three distinct phases of FMDV infection: acute infection (peak viremia), persistent infection (subclinical persistence of FMDV in the upper respiratory tract), and convalescent phase, comprising animals that had successfully cleared the infection (Fig. 1a, b). Distinct miRNA signatures of up- and down-regulated miRNAs in circulation in response to FMDV infection were detected. The results obtained from this proof-of-concept study reinforce the application of this approach to future large scale testing and the investigation of miRNA signatures for other serotypes of FMDV.Table 1

View Article: PubMed Central - PubMed

ABSTRACT

Background: Changes in the levels of circulating microRNAs (miRNAs) in the serum of humans and animals have been detected as a result of infection with a variety of viruses. However, to date, such a miRNA profiling study has not been conducted for foot-and-mouth disease virus (FMDV) infection.

Methods: The relative abundance of 169 miRNAs was measured in bovine serum collected at three different phases of FMDV infection in a proof-of-concept study using miRNA PCR array plates.

Results: Alterations in specific miRNA levels were detected in serum during acute, persistent, and convalescent phases of FMDV infection. Subclinical FMDV persistence produced a circulating miRNA profile distinct from cattle that had cleared infection. bta-miR-17-5p was highest expressed during acute infection, whereas bta-miR-31 was the highest during FMDV persistence. Interestingly, miR-1281was significantly down-regulated during both acute and persistent infection. Cattle that cleared infection resembled the baseline profile, adding support to applying serum miRNA profiling for identification of sub-clinically infected FMDV carriers. Significantly regulated miRNAs during acute or persistent infection were associated with cellular proliferation, apoptosis, modulation of the immune response, and lipid metabolism.

Conclusions: These findings suggest a role for non-coding regulatory RNAs in FMDV infection of cattle. Future studies will delineate the individual contributions of the reported miRNAs to FMDV replication, determine if this miRNA signature is applicable across all FMDV serotypes, and may facilitate development of novel diagnostic applications.

Electronic supplementary material: The online version of this article (doi:10.1186/s12985-017-0743-3) contains supplementary material, which is available to authorized users.

No MeSH data available.


Related in: MedlinePlus