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Identification and expression analysis of the interferon-induced protein with tetratricopeptide repeats 5 (IFIT5) gene in duck (Anas platyrhynchos domesticus).

Wang B, Chen Y, Mu C, Su Y, Liu R, Huang Z, Li Y, Yu Q, Chang G, Xu Q, Chen G - PLoS ONE (2015)

Bottom Line: Based on the sequence obtained, we performed a series of bioinformatics analyses, and found that duIFIT5 was most similar to homologs in other avian species.Finally, we used duck hepatitis virus type 1 (DHV-1) and polyriboinosinicpolyribocytidylic acid (poly (I:C)) as a pathogen or a pathogen-associated molecular pattern induction to infect three-day-old domestic ducklings.Taken together, these results provide a greater understanding of avian IFIT5.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Animal Genetics & Breeding and Molecular Design of Jiangsu province, Yangzhou University, Yangzhou, People's Republic of China.

ABSTRACT
The interferon-induced proteins with tetratricopeptide repeats (IFITs) protein family mediates antiviral effects by inhibiting translation initiation, cell proliferation, and migration in the interferon (IFN) dependent innate immune system. Several members of this family, including IFIT1, IFIT2, IFIT3 and IFIT5, have been heavily studied in mammals. Avian species contain only one family member, IFIT5, and little is known about the role of this protein in birds. In this study, duck IFIT5 (duIFIT5) full-length mRNA was cloned by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of the cDNA ends (RACE). Based on the sequence obtained, we performed a series of bioinformatics analyses, and found that duIFIT5 was most similar to homologs in other avian species. Also, duIFIT5 contained eight conserved TPR motifs and two conserved multi-domains (TPR_11 and TPR_12). Finally, we used duck hepatitis virus type 1 (DHV-1) and polyriboinosinicpolyribocytidylic acid (poly (I:C)) as a pathogen or a pathogen-associated molecular pattern induction to infect three-day-old domestic ducklings. The liver and spleen were collected to detect the change in duIFIT5 transcript level upon infection by quantitative real-time PCR (qRT-PCR). DuIFIT5 expression rapidly increased after DHV-1 infection and maintained a high level, while the transcripts of duIFIT5 peaked at 8h after poly (I:C) infection and then returned to normal. Taken together, these results provide a greater understanding of avian IFIT5.

No MeSH data available.


Related in: MedlinePlus

Relative expression of duIFIT5 in liver (A) and spleen (B) after poly (I:C) injection.qRT-PCR was used to determine the relative expression of duIFIT5 in liver and spleen tissues at 0, 4, 8, 12, 24, 36, 48, 72 and 96 h after infection with poly (I:C). The expression of duIFIT5 was normalized to GAPDH. Different letter showed significant difference (p < 0.05).
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pone.0121065.g006: Relative expression of duIFIT5 in liver (A) and spleen (B) after poly (I:C) injection.qRT-PCR was used to determine the relative expression of duIFIT5 in liver and spleen tissues at 0, 4, 8, 12, 24, 36, 48, 72 and 96 h after infection with poly (I:C). The expression of duIFIT5 was normalized to GAPDH. Different letter showed significant difference (p < 0.05).

Mentions: Upon infection of ducklings with DHV-1, the major pathological change is hepatitis. The same situation also appeared in the dead ducklings of our experiment. In addition, the spleen is an important organ for immune system function. Therefore, liver and spleen tissues of treated and untreated groups were collected to study the temporal expression of duIFIT5 with injection of foreign RNA. The mRNA expression levels of duIFIT5 in liver and spleen were measured at various time points after injection (0, 4, 8, 12, 24, 36, 48, 72, 96 h) by qRT-PCR. For animals infected with DHV-1, some subtle differences were observed in the expression patterns of the spleen compared to the liver. In the liver, duIFIT5 expression sharply increased more than 200- fold from 0–48 hours post-infection and expression levels remained high compared with that of the control for the duration of the experiment. In the spleen, duIFIT5 expression peaked at 36 h post-infection, but only demonstrated a 40-fold increase with that of the control (Fig. 5). After poly (I:C) injection, duIFIT5 expression rapidly increased in both liver and spleen during the first eight hours post infection and reached a peak of 10–20 fold with that of that control, then returned to normal levels by 36 h post induction (Fig. 6). These results showed that duIFIT5 sharply increased following both types of viral infection, but the temporal expression patterns of DHV-1 group and poly (I:C) group were different. The expression pattern of DHV-1group was similar between the liver and spleen, but duIFIT5 shown a stronger increase in liver.


Identification and expression analysis of the interferon-induced protein with tetratricopeptide repeats 5 (IFIT5) gene in duck (Anas platyrhynchos domesticus).

Wang B, Chen Y, Mu C, Su Y, Liu R, Huang Z, Li Y, Yu Q, Chang G, Xu Q, Chen G - PLoS ONE (2015)

Relative expression of duIFIT5 in liver (A) and spleen (B) after poly (I:C) injection.qRT-PCR was used to determine the relative expression of duIFIT5 in liver and spleen tissues at 0, 4, 8, 12, 24, 36, 48, 72 and 96 h after infection with poly (I:C). The expression of duIFIT5 was normalized to GAPDH. Different letter showed significant difference (p < 0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4376821&req=5

pone.0121065.g006: Relative expression of duIFIT5 in liver (A) and spleen (B) after poly (I:C) injection.qRT-PCR was used to determine the relative expression of duIFIT5 in liver and spleen tissues at 0, 4, 8, 12, 24, 36, 48, 72 and 96 h after infection with poly (I:C). The expression of duIFIT5 was normalized to GAPDH. Different letter showed significant difference (p < 0.05).
Mentions: Upon infection of ducklings with DHV-1, the major pathological change is hepatitis. The same situation also appeared in the dead ducklings of our experiment. In addition, the spleen is an important organ for immune system function. Therefore, liver and spleen tissues of treated and untreated groups were collected to study the temporal expression of duIFIT5 with injection of foreign RNA. The mRNA expression levels of duIFIT5 in liver and spleen were measured at various time points after injection (0, 4, 8, 12, 24, 36, 48, 72, 96 h) by qRT-PCR. For animals infected with DHV-1, some subtle differences were observed in the expression patterns of the spleen compared to the liver. In the liver, duIFIT5 expression sharply increased more than 200- fold from 0–48 hours post-infection and expression levels remained high compared with that of the control for the duration of the experiment. In the spleen, duIFIT5 expression peaked at 36 h post-infection, but only demonstrated a 40-fold increase with that of the control (Fig. 5). After poly (I:C) injection, duIFIT5 expression rapidly increased in both liver and spleen during the first eight hours post infection and reached a peak of 10–20 fold with that of that control, then returned to normal levels by 36 h post induction (Fig. 6). These results showed that duIFIT5 sharply increased following both types of viral infection, but the temporal expression patterns of DHV-1 group and poly (I:C) group were different. The expression pattern of DHV-1group was similar between the liver and spleen, but duIFIT5 shown a stronger increase in liver.

Bottom Line: Based on the sequence obtained, we performed a series of bioinformatics analyses, and found that duIFIT5 was most similar to homologs in other avian species.Finally, we used duck hepatitis virus type 1 (DHV-1) and polyriboinosinicpolyribocytidylic acid (poly (I:C)) as a pathogen or a pathogen-associated molecular pattern induction to infect three-day-old domestic ducklings.Taken together, these results provide a greater understanding of avian IFIT5.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Animal Genetics & Breeding and Molecular Design of Jiangsu province, Yangzhou University, Yangzhou, People's Republic of China.

ABSTRACT
The interferon-induced proteins with tetratricopeptide repeats (IFITs) protein family mediates antiviral effects by inhibiting translation initiation, cell proliferation, and migration in the interferon (IFN) dependent innate immune system. Several members of this family, including IFIT1, IFIT2, IFIT3 and IFIT5, have been heavily studied in mammals. Avian species contain only one family member, IFIT5, and little is known about the role of this protein in birds. In this study, duck IFIT5 (duIFIT5) full-length mRNA was cloned by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of the cDNA ends (RACE). Based on the sequence obtained, we performed a series of bioinformatics analyses, and found that duIFIT5 was most similar to homologs in other avian species. Also, duIFIT5 contained eight conserved TPR motifs and two conserved multi-domains (TPR_11 and TPR_12). Finally, we used duck hepatitis virus type 1 (DHV-1) and polyriboinosinicpolyribocytidylic acid (poly (I:C)) as a pathogen or a pathogen-associated molecular pattern induction to infect three-day-old domestic ducklings. The liver and spleen were collected to detect the change in duIFIT5 transcript level upon infection by quantitative real-time PCR (qRT-PCR). DuIFIT5 expression rapidly increased after DHV-1 infection and maintained a high level, while the transcripts of duIFIT5 peaked at 8h after poly (I:C) infection and then returned to normal. Taken together, these results provide a greater understanding of avian IFIT5.

No MeSH data available.


Related in: MedlinePlus