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. 2022 Jun 27;14(8):e15418. doi: 10.15252/emmm.202115418

Figure 5. Design, construction and in vitro investigation of TPA carrying IL15 .

Figure 5

  • A
    TPA DNA carrying a sequence encoding a mouse IL15IgK whose secretion is under the mouse IgK sp.
  • B
    Investigation of the TPA.IL15 IgK in comparison to the native TPA.IL15 in B16.F1 and CT26.CL25 tumour cells. The TPA DNA constructs were transfected into tumour cells, then the bioactivity of secreted IL15 was assessed by evaluating the effects of media from transfected cells on the proliferation of mouse CTLL‐2. TPA DNA without IL15 (mock) was included as control. Data are representative of one experiment and expressed as mean ± SEM. Experiments were repeated twice and performed in triplicates, and one‐way ANOVA with Tukey's HSD test was used for data analysis.
  • C
    Targeted (RGD4C) and non‐targeted (NT) TPA particles carrying IL15 IgK were produced and used to transduce B16.F1 or CT26.CL25 cells at 0.5 × 106 or 1 × 106 TU/cell. Then secreted IL15 was quantified by ELISA in tumour cell media collected on day 5 post‐transduction. RGD4C.TPA without IL15 IgK (mock) was also included as control. Experiments were repeated twice and shown are the results from a representative experiment. Data are expressed as mean ± SEM, one‐way ANOVA with Tukey's HSD test was used for data analysis.
  • D
    Evaluation of CTLL‐2 proliferation in the presence of media collected from B16.F1 or CT26.CL25 tumour cells after transduction with TPA particles carrying IL15 IgK . Cells treated with a recombinant IL2 cytokine were included as a positive control. Data are representative of one experiment, expressed as mean ± SEM and shown as change in CTLL‐2 number relative to treatment initiation day. Experiments were repeated twice and in triplicates, one‐way ANOVA with Tukey's HSD test was used to analyse the data.

Source data are available online for this figure.