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Expression of HSV-TK.007 variant does not affect vaccinia virus replication specificity and selectivity in tumors in vitro and in vivo ( A) Replication of vaccinia virus expressing HSV-TK.007 in different cancer and normal cells in vitro was similar to the control VV-TK(−) vaccinia. A549 (lung cancer cell line), MDA-MB-231 (breast cancer cell line), COLO 205 (colon cancer cell line), and SAEC (normal primary human small airway <t>epithelial</t> cells) were infected with vaccinia viruses WT (TK.WT), control (VV-TK[−]), HSV-TK.WT and HSV-TK.007 at an MOI of 3. 24 h post-infection, infected cells were harvested, and virus titers were determined by plaque assays. PFU per cell values were determined in three biological replicates. Bars indicate SD. Asterisks indicate statistical significance against the control virus, determined by two-way ANOVA followed by Tukey’s multiple comparisons test: ∗ p < 0.05, ∗∗ p < 0.01. (B) Female mice implanted with MC38 were randomized and treated intravenously with vehicle, control (VV-TK[−]), or HSV-TK.007 1E8 PFU ( n = 8 per group). Normal and tumor tissues were collected 72 h post-treatment. Viral genomes were recovered from tissues, and the expression of the E9L viral gene was measured by RT-qPCR. Bars indicate SEM. Two-way ANOVA followed by Tukey’s multiple comparisons test was performed. No statistically significant differences were observed between VV-TK(−) and VV HSV-TK.007. (C) Same study as B, comparing control (VV-TK[−]) and HSV-TK.007 virus dosed i.v. at 1E8 PFU ( n = 8 per group). Tissues were collected 10 days post-viral treatment. Bars indicate SEM. Two-way ANOVA followed by Tukey’s multiple comparisons test was performed. No statistically significant differences were observed between VV-TK(−) and VV HSV-TK.007.
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Expression of HSV-TK.007 variant does not affect vaccinia virus replication specificity and selectivity in tumors in vitro and in vivo ( A) Replication of vaccinia virus expressing HSV-TK.007 in different cancer and normal cells in vitro was similar to the control VV-TK(−) vaccinia. A549 (lung cancer cell line), MDA-MB-231 (breast cancer cell line), COLO 205 (colon cancer cell line), and SAEC (normal primary human small airway epithelial cells) were infected with vaccinia viruses WT (TK.WT), control (VV-TK[−]), HSV-TK.WT and HSV-TK.007 at an MOI of 3. 24 h post-infection, infected cells were harvested, and virus titers were determined by plaque assays. PFU per cell values were determined in three biological replicates. Bars indicate SD. Asterisks indicate statistical significance against the control virus, determined by two-way ANOVA followed by Tukey’s multiple comparisons test: ∗ p < 0.05, ∗∗ p < 0.01. (B) Female mice implanted with MC38 were randomized and treated intravenously with vehicle, control (VV-TK[−]), or HSV-TK.007 1E8 PFU ( n = 8 per group). Normal and tumor tissues were collected 72 h post-treatment. Viral genomes were recovered from tissues, and the expression of the E9L viral gene was measured by RT-qPCR. Bars indicate SEM. Two-way ANOVA followed by Tukey’s multiple comparisons test was performed. No statistically significant differences were observed between VV-TK(−) and VV HSV-TK.007. (C) Same study as B, comparing control (VV-TK[−]) and HSV-TK.007 virus dosed i.v. at 1E8 PFU ( n = 8 per group). Tissues were collected 10 days post-viral treatment. Bars indicate SEM. Two-way ANOVA followed by Tukey’s multiple comparisons test was performed. No statistically significant differences were observed between VV-TK(−) and VV HSV-TK.007.

Journal: Molecular Therapy Oncology

Article Title: Intravenous oncolytic vaccinia expressing transgenes for enhanced safety, inhibition of shedding, imaging, and systemic cancer immunotherapy

doi: 10.1016/j.omton.2026.201153

Figure Lengend Snippet: Expression of HSV-TK.007 variant does not affect vaccinia virus replication specificity and selectivity in tumors in vitro and in vivo ( A) Replication of vaccinia virus expressing HSV-TK.007 in different cancer and normal cells in vitro was similar to the control VV-TK(−) vaccinia. A549 (lung cancer cell line), MDA-MB-231 (breast cancer cell line), COLO 205 (colon cancer cell line), and SAEC (normal primary human small airway epithelial cells) were infected with vaccinia viruses WT (TK.WT), control (VV-TK[−]), HSV-TK.WT and HSV-TK.007 at an MOI of 3. 24 h post-infection, infected cells were harvested, and virus titers were determined by plaque assays. PFU per cell values were determined in three biological replicates. Bars indicate SD. Asterisks indicate statistical significance against the control virus, determined by two-way ANOVA followed by Tukey’s multiple comparisons test: ∗ p < 0.05, ∗∗ p < 0.01. (B) Female mice implanted with MC38 were randomized and treated intravenously with vehicle, control (VV-TK[−]), or HSV-TK.007 1E8 PFU ( n = 8 per group). Normal and tumor tissues were collected 72 h post-treatment. Viral genomes were recovered from tissues, and the expression of the E9L viral gene was measured by RT-qPCR. Bars indicate SEM. Two-way ANOVA followed by Tukey’s multiple comparisons test was performed. No statistically significant differences were observed between VV-TK(−) and VV HSV-TK.007. (C) Same study as B, comparing control (VV-TK[−]) and HSV-TK.007 virus dosed i.v. at 1E8 PFU ( n = 8 per group). Tissues were collected 10 days post-viral treatment. Bars indicate SEM. Two-way ANOVA followed by Tukey’s multiple comparisons test was performed. No statistically significant differences were observed between VV-TK(−) and VV HSV-TK.007.

Article Snippet: Primary SAEC (Lonza CC-2547, Basel, Switzerland) were maintained in airway epithelial cell basal medium (ATCC PCS-300-030), fully supplemented with components from the bronchial epithelial growth kit (ATCC PCS-300-040).

Techniques: Expressing, Variant Assay, Virus, In Vitro, In Vivo, Control, Infection, Quantitative RT-PCR