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Lamivudine Inhibits the Replication of ALV-J Associated Acutely Transforming Virus and its Helper Virus and Tumor Growth In vitro and In vivo.

Wang Y, Xu S, Li S, Su H, Chang S, Li Y, Sun X, Zhao P, Cui Z - Front Microbiol (2015)

Bottom Line: To study the antiviral effects of lamivudine on avian leukosis virus subgroup J (ALV-J) and its inhibitory effect on the growth of fibrosarcomas caused by acute transforming avian leukosis virus, a series of experiments were performed in chicken embryo fibroblast cultures and 1-day-old chickens inoculated with an acutely transforming viral stock Fu-J (SDAU1005).Furthermore, the effect was dose dependent in the concentration range of 1-4 μg/ml.In chicken experiments, lamivudine could decrease the viral loads of SDAU1005 and Fu-J in the plasma of inoculated chickens, delay the appearance of acute sarcomas, and decrease chicken mortality in the early stage.

View Article: PubMed Central - PubMed

Affiliation: College of Animal Science and Veterinary Medicine, Shandong Agricultural University Tai'an, China.

ABSTRACT
To study the antiviral effects of lamivudine on avian leukosis virus subgroup J (ALV-J) and its inhibitory effect on the growth of fibrosarcomas caused by acute transforming avian leukosis virus, a series of experiments were performed in chicken embryo fibroblast cultures and 1-day-old chickens inoculated with an acutely transforming viral stock Fu-J (SDAU1005). This stock was prepared from an acutely fibrosarcoma of field cases in chicken farms and contained both the replication-defective virus Fu-J carrying v-fps oncogene and its helper virus ALV-J strain SDAU1005. The results from three different assays in cell cultures demonstrated the significant inhibitory effect of lamivudine on the replication of both SDAU1005 and Fu-J viruses. Furthermore, the effect was dose dependent in the concentration range of 1-4 μg/ml. In chicken experiments, lamivudine could decrease the viral loads of SDAU1005 and Fu-J in the plasma of inoculated chickens, delay the appearance of acute sarcomas, and decrease chicken mortality in the early stage. This model may be used to directly evaluate the inhibitory effects of lamivudine on such tumors and to understand the relationship between the replication-defective virus and its helper virus while also assessing tumor processes.

No MeSH data available.


Related in: MedlinePlus

Chicken embryo fibroblast (CEF) cells were infected with primary viral stock and administrated isolated viral stock respectively and maintained in DMEM with or without 1 μg/mL lamivudine for 6 days, and ALV-p27 antigen was measured from cellular supernatant every day (A). Viral RNA was extracted on the 7th day and both helper virus and replication-defective virus were quantitated by real-time PCR. Inhibition ratio of lamivudine was calculated to estimate the inhibitory effect of lamivudine on primary viral stock and isolated viral stock (B). Differences in the expression level were assessed by Student’s t-tests. The error bars represent the SEM. ns, no significant difference (p > 0.05).
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Figure 3: Chicken embryo fibroblast (CEF) cells were infected with primary viral stock and administrated isolated viral stock respectively and maintained in DMEM with or without 1 μg/mL lamivudine for 6 days, and ALV-p27 antigen was measured from cellular supernatant every day (A). Viral RNA was extracted on the 7th day and both helper virus and replication-defective virus were quantitated by real-time PCR. Inhibition ratio of lamivudine was calculated to estimate the inhibitory effect of lamivudine on primary viral stock and isolated viral stock (B). Differences in the expression level were assessed by Student’s t-tests. The error bars represent the SEM. ns, no significant difference (p > 0.05).

Mentions: Chicken embryo fibroblasts were infected with primary viral stock and isolated viral stock, respectively, and maintained in DMEM with 1 μg/ml lamivudine for 6 days. Helper virus and replication-defective virus were quantified by real-time PCR, and the inhibition ratio of lamivudine was calculated. The ELISA results showed that lamivudine could inhibit the replication of both primary virus and administered isolated virus. However, no significance was observed between primary virus multiplication and administered isolated virus multiplication during cell maintenance (Figure 3A). Moreover, the quantitative data also revealed that there was no significance observed between the inhibition ratio of lamivudine on the replication of both helper virus and replication-defective virus (Figure 3B). In addition, the viral sequence coding for reverse transcriptase in the ALV pol gene was amplified from primary virus and isolated virus, and sequences were compared. However, no regular mutations related to drug resistance were observed. Above all, there was no detectable drug resistance mutations that had occurred in the viral stock prepared from subcutaneous tumors collected from chickens infected with primary viral stocks.


Lamivudine Inhibits the Replication of ALV-J Associated Acutely Transforming Virus and its Helper Virus and Tumor Growth In vitro and In vivo.

Wang Y, Xu S, Li S, Su H, Chang S, Li Y, Sun X, Zhao P, Cui Z - Front Microbiol (2015)

Chicken embryo fibroblast (CEF) cells were infected with primary viral stock and administrated isolated viral stock respectively and maintained in DMEM with or without 1 μg/mL lamivudine for 6 days, and ALV-p27 antigen was measured from cellular supernatant every day (A). Viral RNA was extracted on the 7th day and both helper virus and replication-defective virus were quantitated by real-time PCR. Inhibition ratio of lamivudine was calculated to estimate the inhibitory effect of lamivudine on primary viral stock and isolated viral stock (B). Differences in the expression level were assessed by Student’s t-tests. The error bars represent the SEM. ns, no significant difference (p > 0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664723&req=5

Figure 3: Chicken embryo fibroblast (CEF) cells were infected with primary viral stock and administrated isolated viral stock respectively and maintained in DMEM with or without 1 μg/mL lamivudine for 6 days, and ALV-p27 antigen was measured from cellular supernatant every day (A). Viral RNA was extracted on the 7th day and both helper virus and replication-defective virus were quantitated by real-time PCR. Inhibition ratio of lamivudine was calculated to estimate the inhibitory effect of lamivudine on primary viral stock and isolated viral stock (B). Differences in the expression level were assessed by Student’s t-tests. The error bars represent the SEM. ns, no significant difference (p > 0.05).
Mentions: Chicken embryo fibroblasts were infected with primary viral stock and isolated viral stock, respectively, and maintained in DMEM with 1 μg/ml lamivudine for 6 days. Helper virus and replication-defective virus were quantified by real-time PCR, and the inhibition ratio of lamivudine was calculated. The ELISA results showed that lamivudine could inhibit the replication of both primary virus and administered isolated virus. However, no significance was observed between primary virus multiplication and administered isolated virus multiplication during cell maintenance (Figure 3A). Moreover, the quantitative data also revealed that there was no significance observed between the inhibition ratio of lamivudine on the replication of both helper virus and replication-defective virus (Figure 3B). In addition, the viral sequence coding for reverse transcriptase in the ALV pol gene was amplified from primary virus and isolated virus, and sequences were compared. However, no regular mutations related to drug resistance were observed. Above all, there was no detectable drug resistance mutations that had occurred in the viral stock prepared from subcutaneous tumors collected from chickens infected with primary viral stocks.

Bottom Line: To study the antiviral effects of lamivudine on avian leukosis virus subgroup J (ALV-J) and its inhibitory effect on the growth of fibrosarcomas caused by acute transforming avian leukosis virus, a series of experiments were performed in chicken embryo fibroblast cultures and 1-day-old chickens inoculated with an acutely transforming viral stock Fu-J (SDAU1005).Furthermore, the effect was dose dependent in the concentration range of 1-4 μg/ml.In chicken experiments, lamivudine could decrease the viral loads of SDAU1005 and Fu-J in the plasma of inoculated chickens, delay the appearance of acute sarcomas, and decrease chicken mortality in the early stage.

View Article: PubMed Central - PubMed

Affiliation: College of Animal Science and Veterinary Medicine, Shandong Agricultural University Tai'an, China.

ABSTRACT
To study the antiviral effects of lamivudine on avian leukosis virus subgroup J (ALV-J) and its inhibitory effect on the growth of fibrosarcomas caused by acute transforming avian leukosis virus, a series of experiments were performed in chicken embryo fibroblast cultures and 1-day-old chickens inoculated with an acutely transforming viral stock Fu-J (SDAU1005). This stock was prepared from an acutely fibrosarcoma of field cases in chicken farms and contained both the replication-defective virus Fu-J carrying v-fps oncogene and its helper virus ALV-J strain SDAU1005. The results from three different assays in cell cultures demonstrated the significant inhibitory effect of lamivudine on the replication of both SDAU1005 and Fu-J viruses. Furthermore, the effect was dose dependent in the concentration range of 1-4 μg/ml. In chicken experiments, lamivudine could decrease the viral loads of SDAU1005 and Fu-J in the plasma of inoculated chickens, delay the appearance of acute sarcomas, and decrease chicken mortality in the early stage. This model may be used to directly evaluate the inhibitory effects of lamivudine on such tumors and to understand the relationship between the replication-defective virus and its helper virus while also assessing tumor processes.

No MeSH data available.


Related in: MedlinePlus