rabbit anti-vacv polyclonal antibody  (Thermo Fisher)

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: A new MVA ancestor-derived oncolytic vaccinia virus induces immunogenic tumor cell death and robust antitumor immune responses.

doi: 10.1016/j.ymthe.2024.05.014

Figure Lengend Snippet: Figure 3. VACV IOVA induces hallmarks of ICD in cancer cells (A and B) Detection of CALR on the surface of infected cancer cells. Cancer cells were infected with the indicated viruses at a MOI of 5 and, 24 h after infection, CALR was detected by flow cytometry. A fixable viability dye eFluor520 was used to discard cells with a disrupted membrane. (A) Percentage of eFluor520CALR+ cells are plotted ± SD for four different replicates. (B) Representative distribution of the CALR+ population within HeLa cells after infection. (C and D) Release of HMGB1 (C) and ATP (D) to cell supernatants after infection. Cells were infected with the indicated viruses at a MOI of 5 and an ELISA assay was used to detect HMGB1 or luminescent detection kit to detect ATP. Data are plotted as fold change versus levels released after WR/TK infection. *p < 0.05; **p < 0.01 ***p < 0.001.

Article Snippet: Primary antibody for immunohistochemistry was a polyclonal rabbit anti-VACV antibody (1:1000, Ref BP1076, OriGene) and biotinylated secondary antibody was a goat anti-rabbit antibody (1:200, Ref BA1000, Vector).

Techniques: Infection, Cytometry, Membrane, Enzyme-linked Immunosorbent Assay

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: A new MVA ancestor-derived oncolytic vaccinia virus induces immunogenic tumor cell death and robust antitumor immune responses.

doi: 10.1016/j.ymthe.2024.05.014

Figure Lengend Snippet: Figure 4. Replication of VACV IOVA and syncytia formation in tumor models in vivo (A) Flow diagram of the experiment. Nine days before, 5 105 Renca or CT26 cells were subcutaneously im- planted in the flank of BALB/c mice. At day 0, a virus dose of 1 107 PFU was intratumorally injected; mice were sacri- ficed and tumors were harvested 4 days later. (B) Quantifi- cation of virus-driven mCherry-specific fluorescence. Plotted are fluorescence of individual tumors and group means ± SD. (C) Viral titers were determined by plaque assays after tumor homogenization. Titers obtained from each independent tumor are plotted with means ± SD. (D) Detection of VACV by immunohistochemistry in paraffin- embedded CT26 tumors. Larger infected cells compatible with syncytia (black arrow) were detected in IOVA-injected tumors (scale bar, 50 mm). ns, not significant; *p < 0.05.

Article Snippet: Primary antibody for immunohistochemistry was a polyclonal rabbit anti-VACV antibody (1:1000, Ref BP1076, OriGene) and biotinylated secondary antibody was a goat anti-rabbit antibody (1:200, Ref BA1000, Vector).

Techniques: In Vivo, Virus, Injection, Homogenization, Immunohistochemistry, Infection

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: A new MVA ancestor-derived oncolytic vaccinia virus induces immunogenic tumor cell death and robust antitumor immune responses.

doi: 10.1016/j.ymthe.2024.05.014

Figure Lengend Snippet: Figure 5. Antitumor activity of VACV IOVA after intratumoral administration (A) Tumor implantation and treatment plan. C57BL/6 mice harboring subcutaneous B16-F10 tumors were randomized and injected twice (days 0 and 4) with an intratumoral dose of 1 107 PFU of tested viruses. Injection of PBS was used as a control. (B) Tumor volume of individual animals, (C) mean tumor size after treatments, and (D) overall survival are plotted for 5–8 mice/group ± SEM. **p < 0.01; ***p < 0.001.

Article Snippet: Primary antibody for immunohistochemistry was a polyclonal rabbit anti-VACV antibody (1:1000, Ref BP1076, OriGene) and biotinylated secondary antibody was a goat anti-rabbit antibody (1:200, Ref BA1000, Vector).

Techniques: Activity Assay, Tumor Implantation, Injection, Control

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: A new MVA ancestor-derived oncolytic vaccinia virus induces immunogenic tumor cell death and robust antitumor immune responses.

doi: 10.1016/j.ymthe.2024.05.014

Figure Lengend Snippet: Figure 7. Toxicity and antitumor activity of VACV IOVA after systemic administration (A) Body weight change after intravenous administration. Balb/c mice were injected intravenously with 1 108 PFU of WR/TK, IOVA, or IOVA/A56+. PBS was used as a control. Weight profiles of IOVA/A56+ and IOVA-injected mice were similar to those injected with PBS, whereas WR/TK-injected mice lost more than 10% in body weight at day 2 after virus injection. (B–E) IOVA strain improves anti-tumor activity after systemic administration. (B) Tumor implantation and treatment plan. Balb/c-bearing Renca tumors were treated with a single intravenous dose of the tested viruses (1 108 PFU/mouse). (C) Tumor size in individual animals, (D) mean tumor size after treatments, and (E) overall survival are plotted for 5–10 mice/group ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: Primary antibody for immunohistochemistry was a polyclonal rabbit anti-VACV antibody (1:1000, Ref BP1076, OriGene) and biotinylated secondary antibody was a goat anti-rabbit antibody (1:200, Ref BA1000, Vector).

Techniques: Activity Assay, Injection, Control, Virus, Tumor Implantation

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: A new MVA ancestor-derived oncolytic vaccinia virus induces immunogenic tumor cell death and robust antitumor immune responses.

doi: 10.1016/j.ymthe.2024.05.014

Figure Lengend Snippet: Figure 8. Antitumor activity and protective immunity induced by VACV IOVA after repeated high-dose intratumoral administrations (A) Tumor implantation and treatment scheme. Balb/c mice harboring subcutaneous Renca tumors were randomized and injected thrice (on days 0, 3, and 6) with an intratumoral dose of 5 107 PFU of the indicated viruses. Injection of PBS was used as a control. (B) Tumor size in individual animals, (C) mean tumor size after treatments, and (D) overall survival are plotted for 10–13 mice/group ± SEM. (E) Tumor rechallenge of recovered mice with Renca cells results in sustained protection. We implanted 5 105 Renca cells in the contralateral flank of mice that had cleared tumor following IOVA treatment (n = 8). Age-matched naive mice (n = 10) were used as controls. CR, complete responses. **p < 0.01; ***p < 0.001.

Article Snippet: Primary antibody for immunohistochemistry was a polyclonal rabbit anti-VACV antibody (1:1000, Ref BP1076, OriGene) and biotinylated secondary antibody was a goat anti-rabbit antibody (1:200, Ref BA1000, Vector).

Techniques: Activity Assay, Tumor Implantation, Injection, Control