Journal: Cancer Medicine

Article Title: Schedule-dependent therapeutic efficacy of L19mTNF-α and melphalan combined with gemcitabine

doi: 10.1002/cam4.89

Figure Lengend Snippet: Administration schedules of gemcitabine (G) and L19mTNF-α/melphalan (L-M) as single or combined treatments

Article Snippet: WEHI-164 mouse fibrosarcoma (3 × 10 6 ) (ECACC, Sigma-Aldrich, Milan, Italy) and K7M2 mouse osteosarcoma (0.3 × 10 6 ) (American Type Culture Collection, Rockville, MD), all BALB/c origin, were subcutaneously (s.c.) implanted in the left flank of 8- to 10-week-old immunocompetent syngeneic BALB/c mice, purchased from Harlan UK (Oxon, U.K.).

Techniques: In Vivo

Journal: Cancer Medicine

Article Title: Schedule-dependent therapeutic efficacy of L19mTNF-α and melphalan combined with gemcitabine

doi: 10.1002/cam4.89

Figure Lengend Snippet: Therapeutic efficacy of the different schedules of drugs administration. Tumor-free survival curves (%) versus time (days) of WEHI-164 (A and B) and K7M2 (C and D) tumor-bearing mice subjected to G-G (black triangles), L-M (black circles), G-L-M-G (black asterisks), or L-M-G-G (black squares) administration schedules. Data are illustrative of at least 10 mice per treatment group.

Article Snippet: WEHI-164 mouse fibrosarcoma (3 × 10 6 ) (ECACC, Sigma-Aldrich, Milan, Italy) and K7M2 mouse osteosarcoma (0.3 × 10 6 ) (American Type Culture Collection, Rockville, MD), all BALB/c origin, were subcutaneously (s.c.) implanted in the left flank of 8- to 10-week-old immunocompetent syngeneic BALB/c mice, purchased from Harlan UK (Oxon, U.K.).

Techniques: Drug discovery

Journal: Cancer Medicine

Article Title: Schedule-dependent therapeutic efficacy of L19mTNF-α and melphalan combined with gemcitabine

doi: 10.1002/cam4.89

Figure Lengend Snippet: Immune population involved in WEHI-164 and K7M2 tumor eradication in in vivo -depleted mice. Tumor-free survival curves (%) versus time (days) of the WEHI-164 and K7M2 tumor-bearing mice subjected to G-G treatment, in vivo depleted with antibodies direct with CD4 + or CD8 + T cells as described in the Materials and Methods section. The tumor-free survival of groups of five WEHI-164 (A) and K7M2 (B) tumor-bearing mice subjected to G-G treatment (open squares), co-CD4-depleted (black triangles), co-CD8-depleted (black asterisks), or untreated tumor-bearing mice (black diamonds) is indicated. Specific cytolytic activity of the immune splenocytes after WEHI-164 and K7M2 tumor cure and persistence of antitumor memory. Specific lysis (%) of WEHI-164 (C) and K7M2 (D) cells by different E:T ratio of splenocytes from WEHI-164- and K7M2-cured mice at 6 months after G-G (open squares) or L-M-G-G (black diamonds) treatment and after s.c. tumor challenge. The specific lysis is totally inhibited by anti-MHC class I antibodies (open diamonds) and unaffected by anti-MHC class II antibodies (black asterisks). Results are representative of three independent 51 Chromium-release experiments with similar results. (E) The ability of WEHI-164- and K7M2-cured mice subjected to G-G and L-M-G-G treatments to reject the first tumor challenge at different times post cure. The number of challenged mice is indicated in round brackets.

Article Snippet: WEHI-164 mouse fibrosarcoma (3 × 10 6 ) (ECACC, Sigma-Aldrich, Milan, Italy) and K7M2 mouse osteosarcoma (0.3 × 10 6 ) (American Type Culture Collection, Rockville, MD), all BALB/c origin, were subcutaneously (s.c.) implanted in the left flank of 8- to 10-week-old immunocompetent syngeneic BALB/c mice, purchased from Harlan UK (Oxon, U.K.).

Techniques: In Vivo, Activity Assay, Lysis

Journal: Cancer Medicine

Article Title: Schedule-dependent therapeutic efficacy of L19mTNF-α and melphalan combined with gemcitabine

doi: 10.1002/cam4.89

Figure Lengend Snippet: Immunohistochemical assessment of tumor infiltrates. Immunohistochemical assessment of CD4 + T cells, CD8 + T cells, and Gr-1 + CD11b + MDSCs in untreated or treated WEHI-164 (A–C) and K7M2 (D–F) tumor-bearing mice 3 days after the conclusion of all therapeutic protocols. Untreated group of mice received PBS only. Results are expressed as cell number (mean ± SD) per high-magnification microscopic field (HMMF). Data are representative of at least three mice per each treatment group. *** P ≤ 0.001; ** P ≤ 0.01; * P ≤ 0.05.

Article Snippet: WEHI-164 mouse fibrosarcoma (3 × 10 6 ) (ECACC, Sigma-Aldrich, Milan, Italy) and K7M2 mouse osteosarcoma (0.3 × 10 6 ) (American Type Culture Collection, Rockville, MD), all BALB/c origin, were subcutaneously (s.c.) implanted in the left flank of 8- to 10-week-old immunocompetent syngeneic BALB/c mice, purchased from Harlan UK (Oxon, U.K.).

Techniques: Immunohistochemical staining

Journal: Cancer Medicine

Article Title: Schedule-dependent therapeutic efficacy of L19mTNF-α and melphalan combined with gemcitabine

doi: 10.1002/cam4.89

Figure Lengend Snippet: Tumor volume assessment after the different therapeutic treatments. Three days after the term of the all therapeutic protocols, the tumor volumes of WEHI-164 and K7M2 untreated and treated tumor-bearing mice were compared. Untreated mice received PBS only. Tumor volumes (cm 3 ) are expressed as mean ± SD. Data are illustrative of least 10 mice per each treatment group.

Article Snippet: WEHI-164 mouse fibrosarcoma (3 × 10 6 ) (ECACC, Sigma-Aldrich, Milan, Italy) and K7M2 mouse osteosarcoma (0.3 × 10 6 ) (American Type Culture Collection, Rockville, MD), all BALB/c origin, were subcutaneously (s.c.) implanted in the left flank of 8- to 10-week-old immunocompetent syngeneic BALB/c mice, purchased from Harlan UK (Oxon, U.K.).

Techniques:

Journal: Cancer Medicine

Article Title: Schedule-dependent therapeutic efficacy of L19mTNF-α and melphalan combined with gemcitabine

doi: 10.1002/cam4.89

Figure Lengend Snippet: Flow-cytometric assessment of regulatory T cells

Article Snippet: WEHI-164 mouse fibrosarcoma (3 × 10 6 ) (ECACC, Sigma-Aldrich, Milan, Italy) and K7M2 mouse osteosarcoma (0.3 × 10 6 ) (American Type Culture Collection, Rockville, MD), all BALB/c origin, were subcutaneously (s.c.) implanted in the left flank of 8- to 10-week-old immunocompetent syngeneic BALB/c mice, purchased from Harlan UK (Oxon, U.K.).

Techniques:

Journal: Molecular Therapy Oncolytics

Article Title: Local administration of IL-12 with an HC vector results in local and metastatic tumor control in pediatric osteosarcoma

doi: 10.1016/j.omto.2020.11.003

Figure Lengend Snippet: The HCA-EFZP-IL-12 vector as a therapy for osteosarcoma (A). Schematic illustration showing the backbone of the HC-AdV HCA-EFZP-IL-12. The adenovirus encodes mouse IL-12 (mIL-12) with a mifepristone (RU-486)-inducible system for controlled expression of IL-12. (B) Scheme of the experimental schedule. K7M2 murine osteosarcoma cells (5 × 10 5 per tibia) were injected into the tibial tuberosity of BALB/c mice (day −10). Animals were treated twice intratumorally with either a saline solution (control group) or 2 × 10 8 IU of HCA-EFZP-IL-12 (days −5 and −7). 3 days later, both groups were administered mifepristone intraperitoneally for 2 weeks with increasing doses (1 mg/kg on days 0–4, 4 mg/kg on days 7–9, and 8 mg/kg on days 10 and 11). Animals were sacrificed when they presented symptoms of disease, or the tumor volume reached 430 mm 3 . (C) Serum concentrations of IL-12 determined 10 h after mifepristone induction on the indicated days. (D) Determination of IL-12 specificity expression after mifepristone induction. K7M2 cells were engrafted into the tibial tuberosity following the same schedule as (B). Animals were randomized to four groups and serum concentration of IL-12 determined 10 h after mifepristone induction on the indicated days. The groups were as follows: (1) control without mifepristone (PBS−), (2) control plus mifepristone (PBS+), (3) HCA-EFZP-IL-12 without mifepristone (HCA-IL-12−), and (4) HCA-EFZP-IL-12 plus mifepristone (HCA-IL-12+).

Article Snippet: The osteosarcoma cell line K7M2 ( ATCC CRL-836 ) was purchased from the American Type Culture Collection (Sigma-Aldrich).

Techniques: Plasmid Preparation, Expressing, Injection, Saline, Control, Concentration Assay

Journal: Molecular Therapy Oncolytics

Article Title: Local administration of IL-12 with an HC vector results in local and metastatic tumor control in pediatric osteosarcoma

doi: 10.1016/j.omto.2020.11.003

Figure Lengend Snippet: Treatment with the HCA-EFZP-IL-12 vector results in immunological memory in treated animals (A) Rechallenge experiment performed with the long-term survivors identified in Figure 1 D. The long-term survivors in the HCA-EFZP-IL-12-treated group (N = 3) were subjected to rechallenge with K7M2 cells and compared with PBS-treated control mice (N = 5). Kaplan-Meier survival curves were constructed, and the p value was calculated with the log-rank test (p = 0.0123). (B) Analyses of tumor burden development in the PBS group (control group) and HCA-EFZP-IL-12-treated group. Tumor volume in mouse tibias was measured on different days until the end of the experiment. (C) Representative macroscopic H&E images of the tibias and lungs from a long-term survivor. Note that the tissues are completely normal.

Article Snippet: The osteosarcoma cell line K7M2 ( ATCC CRL-836 ) was purchased from the American Type Culture Collection (Sigma-Aldrich).

Techniques: Plasmid Preparation, Control, Construct

Journal: Molecular Therapy Oncolytics

Article Title: Local administration of IL-12 with an HC vector results in local and metastatic tumor control in pediatric osteosarcoma

doi: 10.1016/j.omto.2020.11.003

Figure Lengend Snippet: Local HCA-EFZP-IL-12 administration results in T cell infiltration in the osteosarcoma microenvironment (A) Scheme of the experimental schedule. K7M2 murine osteosarcoma cells (5 × 10 5 per tibia) were injected into the tibial tuberosity of BALB/c mice (day −10). Animals were treated twice intratumorally with either a saline solution (control group) or 2 × 10 8 IU of HCA-EFZP-IL-12 (days −5 and −7). 3 days later, both groups were administered mifepristone intraperitoneally for 2 weeks with increasing doses (1 mg/kg on days 0–4, 4 mg/kg on days 7–9, and 8 mg/kg on days 10 and 11). Animals were sacrificed at day 25 after first mifepristone induction. (B and C) Flow cytometry of T cell populations in tumor-infiltrating lymphocytes (TILs) in tibia (B) or lung (C) of mice bearing K7M2 intratibial tumors at 25 days after IL-12 induction. Bars indicate mean ± SD (N = 4–5). Mann-Whitney test.

Article Snippet: The osteosarcoma cell line K7M2 ( ATCC CRL-836 ) was purchased from the American Type Culture Collection (Sigma-Aldrich).

Techniques: Injection, Saline, Control, Flow Cytometry, MANN-WHITNEY