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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

3D8 scFv-treated C57BL/6 mice demonstrate high survival rates against pseudorabies virus (PRV) infection. (A) Schematic diagram for PRV and 3D8 scFv challenge protocol. M: Mock-treated group; A, B, and C: Groups challenged with PRV at LD50; D–F: Groups challenged with 10 LD50. Different amounts of 3D8 scFv administered to each group, indicated at right. (B) Viability of mouse groups showing correlation with the amount of 3D8 scFv treatment. (a) Effect of 3D8 scFv on the survival rate of LD50 PRV infection groups (A: PBS, B: 5 μg 3D8 scFv, C: 10 μg 3D8 scFv) and (b). Effect of 3D8 scFv on the survival rate of 10 LD50 PRV infection groups (D: PBS, E: 5 μg 3D8 scFv, F: 10 μg 3D8 scFv). Results obtained from 21 mice per group (M group: eight mice) for up to 5 days after PRV infection. (C) Varying degrees of symptom development after PRV infection in the mice groups, as described in (A). Dissection picture of group D shows the most severe dermatitis formation, as indicated by red arrows.
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f3-molce-38-9-773: 3D8 scFv-treated C57BL/6 mice demonstrate high survival rates against pseudorabies virus (PRV) infection. (A) Schematic diagram for PRV and 3D8 scFv challenge protocol. M: Mock-treated group; A, B, and C: Groups challenged with PRV at LD50; D–F: Groups challenged with 10 LD50. Different amounts of 3D8 scFv administered to each group, indicated at right. (B) Viability of mouse groups showing correlation with the amount of 3D8 scFv treatment. (a) Effect of 3D8 scFv on the survival rate of LD50 PRV infection groups (A: PBS, B: 5 μg 3D8 scFv, C: 10 μg 3D8 scFv) and (b). Effect of 3D8 scFv on the survival rate of 10 LD50 PRV infection groups (D: PBS, E: 5 μg 3D8 scFv, F: 10 μg 3D8 scFv). Results obtained from 21 mice per group (M group: eight mice) for up to 5 days after PRV infection. (C) Varying degrees of symptom development after PRV infection in the mice groups, as described in (A). Dissection picture of group D shows the most severe dermatitis formation, as indicated by red arrows.

Mentions: The effect of 3D8 scFv as an antiviral agent against PRV in a mouse model was evaluated at protein injection concentrations of 5 and 10 μg (Fig. 3A). The virus was inoculated i.m. in parallel with an i.p. injection of 3D8 scFv. The mice generally began to exhibit clinical signs of illness 3–5 days post-challenge. Approximately 57% of the group inoculated with the LD50 of the virus without 3D8 scFv (group A) survived, whereas the groups treated with the 3D8 scFv at a concentration of 5 μg (group B) or 10 μg (group C) evidenced survival rates of 81% and 76%, respectively, at the same lethal viral dose. Survival rate dropped significantly to 9% in the group that did not receive 3D8 scFv (group D) when inoculated with a much higher viral titer (10× LD50). However, the 3D8 scFv-treated groups had survival rates of 57% (5 μg; group E) and 47% (10 μg; group F) (Fig. 3B). A typical symptom of PRV infection in each group is shown in Fig. 3C. Lesions in the femoral muscles of mice in group D were induced as a result of viral infection, leading to death of the mice, which represented typical symptoms of the disease (Fig. 3C).


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)

3D8 scFv-treated C57BL/6 mice demonstrate high survival rates against pseudorabies virus (PRV) infection. (A) Schematic diagram for PRV and 3D8 scFv challenge protocol. M: Mock-treated group; A, B, and C: Groups challenged with PRV at LD50; D–F: Groups challenged with 10 LD50. Different amounts of 3D8 scFv administered to each group, indicated at right. (B) Viability of mouse groups showing correlation with the amount of 3D8 scFv treatment. (a) Effect of 3D8 scFv on the survival rate of LD50 PRV infection groups (A: PBS, B: 5 μg 3D8 scFv, C: 10 μg 3D8 scFv) and (b). Effect of 3D8 scFv on the survival rate of 10 LD50 PRV infection groups (D: PBS, E: 5 μg 3D8 scFv, F: 10 μg 3D8 scFv). Results obtained from 21 mice per group (M group: eight mice) for up to 5 days after PRV infection. (C) Varying degrees of symptom development after PRV infection in the mice groups, as described in (A). Dissection picture of group D shows the most severe dermatitis formation, as indicated by red arrows.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4588720&req=5

f3-molce-38-9-773: 3D8 scFv-treated C57BL/6 mice demonstrate high survival rates against pseudorabies virus (PRV) infection. (A) Schematic diagram for PRV and 3D8 scFv challenge protocol. M: Mock-treated group; A, B, and C: Groups challenged with PRV at LD50; D–F: Groups challenged with 10 LD50. Different amounts of 3D8 scFv administered to each group, indicated at right. (B) Viability of mouse groups showing correlation with the amount of 3D8 scFv treatment. (a) Effect of 3D8 scFv on the survival rate of LD50 PRV infection groups (A: PBS, B: 5 μg 3D8 scFv, C: 10 μg 3D8 scFv) and (b). Effect of 3D8 scFv on the survival rate of 10 LD50 PRV infection groups (D: PBS, E: 5 μg 3D8 scFv, F: 10 μg 3D8 scFv). Results obtained from 21 mice per group (M group: eight mice) for up to 5 days after PRV infection. (C) Varying degrees of symptom development after PRV infection in the mice groups, as described in (A). Dissection picture of group D shows the most severe dermatitis formation, as indicated by red arrows.
Mentions: The effect of 3D8 scFv as an antiviral agent against PRV in a mouse model was evaluated at protein injection concentrations of 5 and 10 μg (Fig. 3A). The virus was inoculated i.m. in parallel with an i.p. injection of 3D8 scFv. The mice generally began to exhibit clinical signs of illness 3–5 days post-challenge. Approximately 57% of the group inoculated with the LD50 of the virus without 3D8 scFv (group A) survived, whereas the groups treated with the 3D8 scFv at a concentration of 5 μg (group B) or 10 μg (group C) evidenced survival rates of 81% and 76%, respectively, at the same lethal viral dose. Survival rate dropped significantly to 9% in the group that did not receive 3D8 scFv (group D) when inoculated with a much higher viral titer (10× LD50). However, the 3D8 scFv-treated groups had survival rates of 57% (5 μg; group E) and 47% (10 μg; group F) (Fig. 3B). A typical symptom of PRV infection in each group is shown in Fig. 3C. Lesions in the femoral muscles of mice in group D were induced as a result of viral infection, leading to death of the mice, which represented typical symptoms of the disease (Fig. 3C).

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