Extended Data Fig. 7: Evaluation of ΔXIV biodistribution and detection of checkpoint blockade nanobodies expression in a bilateral tumor model.

a, Schematic of the experimental procedure and schedule for evaluating the biodistribution of ΔXIV in a bilateral A20 lymphoma mouse model. Mice received an intratumoral injection of ΔXIV-lux (5 × 10⁶ c.f.u.) in the left flank tumors when the tumor volume reached ~100 mm³. b, The bioluminescence signal intensity was quantified with an in vivo imaging system on Days 0, 1, 3, 6, 9, and 12 after bacteria injection to track the colonization of ΔXIV in tumors (n = 4 mice per group). c, Bioluminescence quantification of the IVIS imaging in b. d, Schematic illustrating the experimental procedure and schedule for evaluating immune checkpoint blockade nanobodies expression via ΔXIV_PadC4-NETMAP_{PD-L1nb+CTLA-4nb} in a bilateral A20 lymphoma mouse tumor model. A bilateral A20 lymphoma mouse model was established. ΔXIV_PadC4-NETMAP_{PD-L1nb+CTLA-4nb} (5 × 10⁶ c.f.u.) were injected into the left hind flank tumor and exposed to 710 nm light for 2 h daily. Four days after illumination, tumors on both sides were collected and stained for immunofluorescence. e, Immunofluorescence staining of tumor slices. PD-L1nb-Flag tag (red) and CTLA-4nb-HA tag (green) secreted by ΔXIV-NETMAP_{PD-L1nb+CTLA-4nb} were stained with anti-Flag and anti-HA antibodies. Nuclei were stained with DAPI (blue). Three times the experiment was repeated with similar results. Images are representative of 3 mice per group. Scale bar, 25 μm. Data (c) are presented as means ± s.e.m.; n = 4 mice per group. Statistical analyses in c were performed using a two-tailed unpaired t-test.

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