Volume 3, Issue 2, Pages 160-168 (February 2001) Development of a Syngenic Murine B16 Cell Line-Derived Melanoma Susceptible to Destruction by Neuroattenuated HSV-1  Cathie G. Miller, Claude Krummenacher, Rosalyn J. Eisenberg, Gary H. Cohen, Nigel W. Fraser  Molecular Therapy  Volume 3, Issue 2, Pages 160-168 (February 2001) DOI: 10.1006/mthe.2000.0240 Copyright © 2001 American Society for Gene Therapy Terms and Conditions

FIG. 1 HSV-1 17 + efficiently replicates in and kills A10 and C10 cells, but not B5 or control cells. Subconfluent cell cultures were infected with wild-type HSV-1 17+ with different multiplicities of infection (m.o.i.) as described under Methods and percentages of surviving cells were recorded at 24, 48, and 72 h. These data are representative of two separate experiments. (A) m.o.i. 0.01, (B) m.o.i. 0.1, (C) m.o.i. 1.0. Molecular Therapy 2001 3, 160-168DOI: (10.1006/mthe.2000.0240) Copyright © 2001 American Society for Gene Therapy Terms and Conditions

FIG. 2 HSV-1 17+ efficiently replicates in and kills A10 and C10 cells, but not B5 or control cells. Subconfluent cell cultures were infected with HSV-1 17+ with different multiplicities of infection (m.o. i.) as described under Methods and percentages of surviving cells were recorded at 24, 48, and 72 h. These data are representative of two separate experiments. (A) m.o.i. 0.01, (B) m.o.i. 0.1, (C) m.o.i. 1.0. Molecular Therapy 2001 3, 160-168DOI: (10.1006/mthe.2000.0240) Copyright © 2001 American Society for Gene Therapy Terms and Conditions

FIG. 3 Production of tumors in vivo after subcutaneous injection of 5 × 106 Hve-transfected cells. (A) Primary tumor growth. All mice injected with A10, C10, and control cells grew tumors. Only 33% of the mice injected with B5 cells grew tumors (N = 10 for A10, B5, and control cells; N = 15 for C10 cells). (B) Secondary tumor growth. After allowing growth of primary tumors in mice for 6 weeks, tumor cells were injected into opposite flanks of mice that grew primary tumors. All A10, C10, and control cells injected into mice grew secondary tumors. Only 33% of the B5-injected mice grew secondary tumors (N = 10 for A10 and control cells, N = 15 for C10, and N = 3 for B5). Molecular Therapy 2001 3, 160-168DOI: (10.1006/mthe.2000.0240) Copyright © 2001 American Society for Gene Therapy Terms and Conditions

FIG. 4 Survival of C57BI/6 mice after intracranial injection of 5 × 104 Hve-transfected cells followed by treatment injection of 5 × 104 pfu HSV-1 1716 on day 10. (A) A10. There is a significant increase in survival of mice receiving viral treatment compared to those receiving mock treatment, P = 0.002. N = 10 for HSV-1 1716 and mock treated. (B) B5. There is no significant increase in survival of mice receiving viral treatment compared to those receiving mock treatment. N = 5 for HSV-1 1716 and mock treated. (C) C10. There is a significant increase in survival of mice receiving viral treatment compared to those receiving mock treatment, P < 0.0001. (D) Control cells. There is no significant increase in survival of mice receiving viral treatment compared to those receiving mock treatment, P = 0.313. N = 10 for HSV-1 1716 and mock treated. Molecular Therapy 2001 3, 160-168DOI: (10.1006/mthe.2000.0240) Copyright © 2001 American Society for Gene Therapy Terms and Conditions