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Therapeutic Strategy for the Prevention of Pseudorabies Virus Infection in C57BL/6 Mice by 3D8 scFv with Intrinsic Nuclease Activity.

Lee G, Cho S, Hoang PM, Kim D, Lee Y, Kil EJ, Byun SJ, Lee TK, Kim DH, Kim S, Lee S - Mol. Cells (2015)

Bottom Line: The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays.Intraperitoneal injection of 5 and 10 μg 3D8 scFv resulted in no detectable toxicity.The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Korea.

ABSTRACT
3D8 single chain variable fragment (scFv) is a recombinant monoclonal antibody with nuclease activity that was originally isolated from autoimmune-prone MRL mice. In a previous study, we analyzed the nuclease activity of 3D8 scFv and determined that a HeLa cell line expressing 3D8 scFv conferred resistance to herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PRV). In this study, we demonstrate that 3D8 scFv could be delivered to target tissues and cells where it exerted a therapeutic effect against PRV. PRV was inoculated via intramuscular injection, and 3D8 scFv was injected intraperitoneally. The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays. Intraperitoneal injection of 5 and 10 μg 3D8 scFv resulted in no detectable toxicity. The survival rate in C57BL/6 mice was 9% after intramuscular injection of 10 LD50 PRV. In contrast, the 3D8 scFv-injected C57BL/6 mice showed survival rates of 57% (5 μg) and 47% (10 μg). The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.

No MeSH data available.


Related in: MedlinePlus

The antiviral resistance observed in the 3D8 scFv-treated mice is not dependent on chemokines. Quantitative real-time reverse transcription-polymerase chain reaction analysis of the chemokine-related genes in the pseudorabies virus (PRV)-infected mice treated with (groups E and F) or without 3D8 scFv (group D). (A) Inducible nitric oxide synthase (iNOS) and C-X-C motif chemokine 10 (CXCL10) mRNA expression in muscle of mice following PRV infection. (B) iNOS and CXCL10 mRNA expression in the brains of mice following PRV infection. Data are mean ± standard error. ***Significantly different from group D at p < 0.001 (one-way analysis of variance and Tukey’s post hoc t-test). M: Mock-treated group; D challenged with 10LD50 PRV followed by PBS treatment; E challenged with 10LD50 PRV followed by treatment with 5 μg of 3D8 scFv; F challenged with 10LD50 PRV followed by 10 μg of 3D8 scFv treatment.
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f5-molce-38-9-773: The antiviral resistance observed in the 3D8 scFv-treated mice is not dependent on chemokines. Quantitative real-time reverse transcription-polymerase chain reaction analysis of the chemokine-related genes in the pseudorabies virus (PRV)-infected mice treated with (groups E and F) or without 3D8 scFv (group D). (A) Inducible nitric oxide synthase (iNOS) and C-X-C motif chemokine 10 (CXCL10) mRNA expression in muscle of mice following PRV infection. (B) iNOS and CXCL10 mRNA expression in the brains of mice following PRV infection. Data are mean ± standard error. ***Significantly different from group D at p < 0.001 (one-way analysis of variance and Tukey’s post hoc t-test). M: Mock-treated group; D challenged with 10LD50 PRV followed by PBS treatment; E challenged with 10LD50 PRV followed by treatment with 5 μg of 3D8 scFv; F challenged with 10LD50 PRV followed by 10 μg of 3D8 scFv treatment.

Mentions: The expression levels of genes involved in the inflammation response were assessed via qRT-PCR with RNA samples obtained from the groups described in Fig. 3A to determine whether the antiviral mechanisms of 3D8 scFv include any indirect effects such as triggering of endogenous antiviral responses. As demonstrated in Fig. 5, the levels of two transcripts associated with the inflammation response, CXCL10 and iNOS, were detected at elevated levels in the PRV-infected samples (groups D–F). However, no significant differences in the expression levels of these transcripts were observed between the survivors and dead among these groups, thereby confirming the absence of any indirect effect of the antiviral mechanism exploited by 3D8 scFv (Figs. 5A and 5B).


Therapeutic Strategy for the Prevention of Pseudorabies Virus Infection in C57BL/6 Mice by 3D8 scFv with Intrinsic Nuclease Activity.

Lee G, Cho S, Hoang PM, Kim D, Lee Y, Kil EJ, Byun SJ, Lee TK, Kim DH, Kim S, Lee S - Mol. Cells (2015)

The antiviral resistance observed in the 3D8 scFv-treated mice is not dependent on chemokines. Quantitative real-time reverse transcription-polymerase chain reaction analysis of the chemokine-related genes in the pseudorabies virus (PRV)-infected mice treated with (groups E and F) or without 3D8 scFv (group D). (A) Inducible nitric oxide synthase (iNOS) and C-X-C motif chemokine 10 (CXCL10) mRNA expression in muscle of mice following PRV infection. (B) iNOS and CXCL10 mRNA expression in the brains of mice following PRV infection. Data are mean ± standard error. ***Significantly different from group D at p < 0.001 (one-way analysis of variance and Tukey’s post hoc t-test). M: Mock-treated group; D challenged with 10LD50 PRV followed by PBS treatment; E challenged with 10LD50 PRV followed by treatment with 5 μg of 3D8 scFv; F challenged with 10LD50 PRV followed by 10 μg of 3D8 scFv treatment.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4588720&req=5

f5-molce-38-9-773: The antiviral resistance observed in the 3D8 scFv-treated mice is not dependent on chemokines. Quantitative real-time reverse transcription-polymerase chain reaction analysis of the chemokine-related genes in the pseudorabies virus (PRV)-infected mice treated with (groups E and F) or without 3D8 scFv (group D). (A) Inducible nitric oxide synthase (iNOS) and C-X-C motif chemokine 10 (CXCL10) mRNA expression in muscle of mice following PRV infection. (B) iNOS and CXCL10 mRNA expression in the brains of mice following PRV infection. Data are mean ± standard error. ***Significantly different from group D at p < 0.001 (one-way analysis of variance and Tukey’s post hoc t-test). M: Mock-treated group; D challenged with 10LD50 PRV followed by PBS treatment; E challenged with 10LD50 PRV followed by treatment with 5 μg of 3D8 scFv; F challenged with 10LD50 PRV followed by 10 μg of 3D8 scFv treatment.
Mentions: The expression levels of genes involved in the inflammation response were assessed via qRT-PCR with RNA samples obtained from the groups described in Fig. 3A to determine whether the antiviral mechanisms of 3D8 scFv include any indirect effects such as triggering of endogenous antiviral responses. As demonstrated in Fig. 5, the levels of two transcripts associated with the inflammation response, CXCL10 and iNOS, were detected at elevated levels in the PRV-infected samples (groups D–F). However, no significant differences in the expression levels of these transcripts were observed between the survivors and dead among these groups, thereby confirming the absence of any indirect effect of the antiviral mechanism exploited by 3D8 scFv (Figs. 5A and 5B).

Bottom Line: The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays.Intraperitoneal injection of 5 and 10 μg 3D8 scFv resulted in no detectable toxicity.The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Korea.

ABSTRACT
3D8 single chain variable fragment (scFv) is a recombinant monoclonal antibody with nuclease activity that was originally isolated from autoimmune-prone MRL mice. In a previous study, we analyzed the nuclease activity of 3D8 scFv and determined that a HeLa cell line expressing 3D8 scFv conferred resistance to herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PRV). In this study, we demonstrate that 3D8 scFv could be delivered to target tissues and cells where it exerted a therapeutic effect against PRV. PRV was inoculated via intramuscular injection, and 3D8 scFv was injected intraperitoneally. The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays. Intraperitoneal injection of 5 and 10 μg 3D8 scFv resulted in no detectable toxicity. The survival rate in C57BL/6 mice was 9% after intramuscular injection of 10 LD50 PRV. In contrast, the 3D8 scFv-injected C57BL/6 mice showed survival rates of 57% (5 μg) and 47% (10 μg). The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.

No MeSH data available.


Related in: MedlinePlus