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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

Inhibitory effects of lamivudine in different concentrations on replication of Fu-J and its helper virus in CEF cells infected with Fu-1 stock. (A) Comparisons of ALV p27 antigen levels in cell culture supernatants by ELASA. The vertical axis represents the s/p values in ELISA, the cut-off value of positive criteria was 0.2. (B) Comparisons of expression level of genomic RNA fragments gp85 specific to SDAU1005 and fps to Fu-J by real-time PCR. The expression levels were normalized with the expression level of chicken β-actin mRNA and performed by using the 2-ΔΔCT method. Differences in the expression level were assessed by Student’s t-tests. (A,B) Differences were considered significant when p < 0.01 (∗), highly significant when p ≤ 0.01 (∗∗) and extremely significant p ≤ 0.001 (∗∗∗). The error bars represent the SEM. The data are based on the results of three independent experiments. (C) Detection of ALV-J gp85 and fps proteins in Fu-J (SDAU1005) viral stock infected CEF cells by western blot analysis. Line 1: control group; lines 2–4: cells treated with 1, 2, 4 μg/mL lamivudine.
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Figure 1: Inhibitory effects of lamivudine in different concentrations on replication of Fu-J and its helper virus in CEF cells infected with Fu-1 stock. (A) Comparisons of ALV p27 antigen levels in cell culture supernatants by ELASA. The vertical axis represents the s/p values in ELISA, the cut-off value of positive criteria was 0.2. (B) Comparisons of expression level of genomic RNA fragments gp85 specific to SDAU1005 and fps to Fu-J by real-time PCR. The expression levels were normalized with the expression level of chicken β-actin mRNA and performed by using the 2-ΔΔCT method. Differences in the expression level were assessed by Student’s t-tests. (A,B) Differences were considered significant when p < 0.01 (∗), highly significant when p ≤ 0.01 (∗∗) and extremely significant p ≤ 0.001 (∗∗∗). The error bars represent the SEM. The data are based on the results of three independent experiments. (C) Detection of ALV-J gp85 and fps proteins in Fu-J (SDAU1005) viral stock infected CEF cells by western blot analysis. Line 1: control group; lines 2–4: cells treated with 1, 2, 4 μg/mL lamivudine.

Mentions: Figure 1 shows that significant inhibitory effects of lamivudine occurred with respect to replication of both Fu-J and its helper virus SDAU1005 in infected CEF cultures. This was based on different criteria in three different assays compared with the control without lamivudine. These criteria and assay included ALV-p27 expression levels in cell culture supernatants by ELSA kit (Figure 1A), specific viral genomic RNA fragment levels in cell culture supernatants by real-time RT-PCR (Figure 1B), and specific gp85 or fps protein expression levels in cell lysates by western blot analysis (Figure 1C). Such inhibitory effects of lamivudine were dose dependent in the concentration range from 1 to 4 μg/ml as indicated in Figure 1. To rule out the possibility that the antiviral activity was due to the cytotoxicity of the chemical, a CCK-8 cell viability assay was performed, and this indicated that there was no toxic effect of lamivudine on CEF viability with the concentration used (data was not included), while lamivudine with a concentration exceeding 8 μg/ml resulted in mild damage to CEF.


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)

Inhibitory effects of lamivudine in different concentrations on replication of Fu-J and its helper virus in CEF cells infected with Fu-1 stock. (A) Comparisons of ALV p27 antigen levels in cell culture supernatants by ELASA. The vertical axis represents the s/p values in ELISA, the cut-off value of positive criteria was 0.2. (B) Comparisons of expression level of genomic RNA fragments gp85 specific to SDAU1005 and fps to Fu-J by real-time PCR. The expression levels were normalized with the expression level of chicken β-actin mRNA and performed by using the 2-ΔΔCT method. Differences in the expression level were assessed by Student’s t-tests. (A,B) Differences were considered significant when p < 0.01 (∗), highly significant when p ≤ 0.01 (∗∗) and extremely significant p ≤ 0.001 (∗∗∗). The error bars represent the SEM. The data are based on the results of three independent experiments. (C) Detection of ALV-J gp85 and fps proteins in Fu-J (SDAU1005) viral stock infected CEF cells by western blot analysis. Line 1: control group; lines 2–4: cells treated with 1, 2, 4 μg/mL lamivudine.
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Figure 1: Inhibitory effects of lamivudine in different concentrations on replication of Fu-J and its helper virus in CEF cells infected with Fu-1 stock. (A) Comparisons of ALV p27 antigen levels in cell culture supernatants by ELASA. The vertical axis represents the s/p values in ELISA, the cut-off value of positive criteria was 0.2. (B) Comparisons of expression level of genomic RNA fragments gp85 specific to SDAU1005 and fps to Fu-J by real-time PCR. The expression levels were normalized with the expression level of chicken β-actin mRNA and performed by using the 2-ΔΔCT method. Differences in the expression level were assessed by Student’s t-tests. (A,B) Differences were considered significant when p < 0.01 (∗), highly significant when p ≤ 0.01 (∗∗) and extremely significant p ≤ 0.001 (∗∗∗). The error bars represent the SEM. The data are based on the results of three independent experiments. (C) Detection of ALV-J gp85 and fps proteins in Fu-J (SDAU1005) viral stock infected CEF cells by western blot analysis. Line 1: control group; lines 2–4: cells treated with 1, 2, 4 μg/mL lamivudine.
Mentions: Figure 1 shows that significant inhibitory effects of lamivudine occurred with respect to replication of both Fu-J and its helper virus SDAU1005 in infected CEF cultures. This was based on different criteria in three different assays compared with the control without lamivudine. These criteria and assay included ALV-p27 expression levels in cell culture supernatants by ELSA kit (Figure 1A), specific viral genomic RNA fragment levels in cell culture supernatants by real-time RT-PCR (Figure 1B), and specific gp85 or fps protein expression levels in cell lysates by western blot analysis (Figure 1C). Such inhibitory effects of lamivudine were dose dependent in the concentration range from 1 to 4 μg/ml as indicated in Figure 1. To rule out the possibility that the antiviral activity was due to the cytotoxicity of the chemical, a CCK-8 cell viability assay was performed, and this indicated that there was no toxic effect of lamivudine on CEF viability with the concentration used (data was not included), while lamivudine with a concentration exceeding 8 μg/ml resulted in mild damage to CEF.

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