Journal: Cancer Immunology, Immunotherapy

Article Title: Antitumor in situ vaccination effect of TNFα and IL-12 plasmid DNA electrotransfer in a murine melanoma model

doi: 10.1007/s00262-018-2133-0

Figure Lengend Snippet: Experimental design. The tested therapeutic groups were as follows: TNF, gene electrotransfer (GET) of the TNFα plasmid; IL-12 GET, GET of the IL-12 plasmid repeated twice with an interval of 6 days; and TNF + IL-12, concomitant GET of the TNFα and IL-12 plasmids, followed 6 days later by GET of the IL-12 plasmid. Additional control groups were as follows: CTRL complete control, EP electroporation only, pControl GET of a control plasmid. On days 2 and 8 tumors were harvested for determination of TNFα and IL-12 expression by RT-PCR. Tumor growth was monitored until the tumor reached a volume of 300 mm 3 , which was also used as the endpoint event for plotting the Kaplan–Meier survival curve. Mice that were tumor free for 90 days were subjected to secondary challenge with an injection of tumor cells. Tumors, blood and lymph nodes were collected on days 4 and 10. Tumors were used for histological determination of immune cell infiltration (H&E, hematoxylin and eosin staining) and immunohistochemical (IHC) determination of granzyme B-positive cells. Granzyme B ELISpot was performed on the PBMC and single-cell suspensions isolated from the lymph nodes

Article Snippet: A Mouse Granzyme B ELISpot kit (R&D Systems, Minneapolis, MN, USA) was used to detect granzyme B-positive cells in blood and lymph node samples.

Techniques: Electrotransfer, Plasmid Preparation, Control, Electroporation, Expressing, Reverse Transcription Polymerase Chain Reaction, Injection, Staining, Immunohistochemical staining, Enzyme-linked Immunospot, Isolation

Journal: Cancer Immunology, Immunotherapy

Article Title: Antitumor in situ vaccination effect of TNFα and IL-12 plasmid DNA electrotransfer in a murine melanoma model

doi: 10.1007/s00262-018-2133-0

Figure Lengend Snippet: Results of the granzyme B ELISpot performed on single-cell suspensions from the lymph nodes and PBMC 4 days after treatment. a Number of cells isolated from the lymph nodes. b Number of PBMC isolated from blood. c Number of granzyme B-positive cells in the lymph nodes after exposure to tumor cells, normalized to cells that were not stimulated with tumor cells. d Number of granzyme B-positive PBMC after exposure to tumor cells, normalized to cells that were not stimulated with tumor cells. Circles represent values for individual samples; boxes show the variance with the median values for three samples per experimental group. CTRL control group, EP application of electroporation only, TNF TNFα GET, IL-12 IL-12 GET, TNF + IL-12 TNFα and IL-12 GET

Article Snippet: A Mouse Granzyme B ELISpot kit (R&D Systems, Minneapolis, MN, USA) was used to detect granzyme B-positive cells in blood and lymph node samples.

Techniques: Enzyme-linked Immunospot, Isolation, Control, Electroporation