Establishment of a novel therapeutic vector targeting the trigeminal ganglion in rats

Figures & data

Figure 1 Construction of the rdHSV-IFNγ therapeutic system.

Notes: (A) HSV backbone of the recombined HSV-1. (B) Shuttle plasmids carrying the IFNγ gene. (C) The complete rdHSV-1-IFNγ therapeutic system.
Abbreviations: HSV-1, herpes simplex virus type 1; IFNγ, interferon gamma; rdHSV, replication-defective HSV-1; EF-1α, elongation factor 1 alpha; CMV, cytomegalovirus promoter.

Figure 2 Safety evaluation of rdHSV-IFNγ in vivo and in vitro.

Notes: (A) Immunization procedure for the rdHSV-IFNγ vector. Corneas were scarified ten times with syringe needle (1 mL type), and drops of rdHSV-IFNγ solution were applied to the ocular mucosa. (B) Kaplan–Meier survival analysis for the three treatment groups. (C) Infection rates in the three treatment groups. (D) CC₅₀ of 0.1% DMSO, aciclovir, and rdHSV-IFNγ. ***P<0.001, CC₅₀, or EC₅₀ in the rdHSV-IFNγ group vs the aciclovir group.
Abbreviations: CC₅₀, 50% cellular cytotoxicity; CN, control; DMSO, dimethyl sulfoxide; EC₅₀, 50% effective concentration; HSV-1, herpes simplex virus type 1; IFNγ, interferon gamma; rdHSV, replication-defective HSV-1.

Figure 3 RT-PCR assay of mRNA expression in the cornea, trigeminal nerve, and TG from day 1 to day 21.

Notes: (A) IFNγ mRNA on day 1 to day 21 post immunization in the cornea, trigeminal nerve, and TG. (B) Statistical analysis for change in IFNγ mRNA expression in the cornea, trigeminal nerve, and TG from day 1 to day 21.
Abbreviations: IFNγ, interferon gamma; RT-PCR, real-time polymerase chain reaction; TG, trigeminal ganglion.

Figure 4 Immunohistochemical assay of IFNγ protein localization in the cornea (A), trigeminal nerve (B), and TG (C) from day 1 to day 21 post immunization.

Notes: The long arrow in the graph illustrates the direction of change in the expression of IFNγ protein. The gray arrow illustrates the rdHSV-IFNγ targeting pathway from cornea to TG.
Abbreviations: HSV-1, herpes simplex virus type 1; IFNγ, interferon gamma; rdHSV, replication-defective HSV-1; TG, trigeminal ganglion.

Figure 5 IFNγ mRNA (A) and protein (B) expression in the TG tissues from day 1 to day 21 post immunization.

Notes: **P<0.01. IFNγ mRNA or protein expression at 21 days vs 14 days.
Abbreviations: IFNγ, interferon gamma; mRNA, messenger RNA; TG, trigeminal ganglion.