vero cells  (ATCC)

Journal: Molecular Therapy Oncology

Article Title: A measles virus encoding a CD19/CD3 bispecific T cell engager shows enhanced preclinical anti-BCP-ALL efficacy without significant toxicity

doi: 10.1016/j.omton.2026.201127

Figure Lengend Snippet: MV-Blina, an oncolytic measles virus encoding the CD19/CD3-bispecific T cell engager, replicates and is cytotoxic (A) Schematic of MV-Blina. Genomes of the parental MV-Edm strain and the recombinant MV-Blina. The bsTE construct contains the anti-human-CD19 variable heavy/light domain (CD19 V H/L ) and the anti-human-CD3 variable heavy/light domain (CD3 V H/L ). (B) Enhanced replication of MV-Blina compared to MV-Edm. Replication of MV-Blina was assessed by infecting Vero cells at MOI 0.01 or 1.0 with MV-Blina and MV-Edm, respectively. Viral titers at the indicated time points were determined with a titration assay. Results are shown as means ± SD of n = 3 from independent experiments seeded in duplicates. (C) Comparable cytotoxicity of MV-Blina and parental MV-Edm. Vero cells were infected with MOI 0.01, 0.1, or 1.0 of MV-Blina and MV-Edm, respectively. Cell viability was determined using the MTT assay at the indicated time points. Results are shown as means ± SD of n ≥ 3 from independent experiments seeded in sextuplicates. Statistical analysis was performed using two-way ANOVA with Dunnett’s correction. ns, not significant; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; and ∗∗∗∗, p < 0.0001.

Article Snippet: The recombinant MV-bsTE virus was generated by transfecting Vero cells with antigenomic cDNA constructs as described previously., The MV-Edmonston B strain (VR-24, ATCC, Manassas, VA, USA) was obtained and amplified prior to experiments.

Techniques: Virus, Recombinant, Construct, Titration, Infection, MTT Assay

Journal: Molecular Therapy Oncology

Article Title: A measles virus encoding a CD19/CD3 bispecific T cell engager shows enhanced preclinical anti-BCP-ALL efficacy without significant toxicity

doi: 10.1016/j.omton.2026.201127

Figure Lengend Snippet: MV-Blina infected cells secrete target-binding secBlina, which initiates ALL cell kill more efficiently than CD19/CD3-bsTE (A) Vero cells secrete secBlina. secBlina was harvested from the supernatant of infected Vero cells, followed by purification and concentration. Eluate (2 μg protein) was detected via His-tag by Western Blot in comparison to the commercial CD19/CD3-bsTE (1.8 μg). (B) Purified and concentrated secBlina binds specifically to target cells. Binding to target cells was confirmed by flow cytometry using CD3 − CD19 + REH, CD3 + CD19 - Jurkat, and CD3 − CD19 - K562 cell lines incubated with secBlina. Binding of secBlina to target cells was detected by flow cytometry via a His-tag antibody. (C) secBlina strongly binds to REH target cells. The REH cell line was first incubated with 1 μg secBlina. Increasing concentrations of CD19/CD3-bsTE were then added to displace cell-bound secBlina. Cell-bound secBlina was detected via an HA tag antibody by flow cytometry. (D) T cells in human PBMCs are activated by secBlina. Pooled PBMCs were stimulated with an (E) T cells are activated by secBlina during co-culture of PBMCs with REH cells. Pooled PBMCs were co-cultured with REH cells in a 1:1-ratio for 24 h while being treated with secBlina or the CD19/CD3-bsTE. Activated CD2 + CD69 + T cells were detected by flow cytometry. (F) MV-Blina infected leukemia cells produce and secrete secBlina. REH and Jurkat cells are shown 72 h post MV infection or Opti-MEM control. secBlina was detected by His-tag IF staining. Scale bars 50 μm. (G) secBlina effectively induces REH target cell kill compared to CD19/CD3-bsTE, whereas K562 control cells are unaffected. Target (T) cells REH (left panels) and negative control cells K562 (right panels) were incubated with secBlina (green) or CD19/CD3-bsTE (blue) for 24 h at indicated concentrations in the presence of PBMCs (effector; E) at different E:T ratios. Specific cell kill was determined by flow cytometry. Results in A, B, C, and F are shown as representatives of three independent experiments. Data are shown as mean ± SD of n = 3 (D, E) or n = 5 (G) independent experiments. Statistical analysis was performed using two-way ANOVA with Dunnett’s correction. ns, not significant; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; and ∗∗∗∗, p < 0.0001.

Article Snippet: The recombinant MV-bsTE virus was generated by transfecting Vero cells with antigenomic cDNA constructs as described previously., The MV-Edmonston B strain (VR-24, ATCC, Manassas, VA, USA) was obtained and amplified prior to experiments.

Techniques: Infection, Binding Assay, Purification, Concentration Assay, Western Blot, Comparison, Flow Cytometry, Incubation, Co-Culture Assay, Cell Culture, Control, Staining, Negative Control