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. Author manuscript; available in PMC: 2020 Jan 3.
Published in final edited form as: Nat Med. 2019 Jul 3;25(7):1057–1063. doi: 10.1038/s41591-019-0498-z

Figure 1 |. Quorum-induced release of functional anti-CD47 blocking nanobody by engineered immunotherapeutic bacteria encoding a synchronized lysis circuit (SLC).

Figure 1 |

a, E. coli with SLC reach a quorum and induce the phage lysis protein ϕX174E, leading to bacterial lysis and release of a constitutively produced, anti-CD47 blocking nanobody which binds to CD47 on the tumor cell surface. b, Bacterial growth dynamics over time of SLC+ and SLC− E. coli in batch liquid culture. Data are representative of three independent experimental replicates c, A20 cells were co-incubated with constant concentration of FITC conjugated αCD47 monoclonal antibody (FITC-miap301) along with varying concentrations of bacterial lysates containing constitutively expressed CD47nb (pSC02) or empty vector (pSC03). Data are representative of two independent experimental replicates d, in vitro phagocytosis of DiI labeled A20 cells pretreated with PBS, SLC+ bacteria lysate or SLC+ CD47nb+ bacteria lysate by bone-marrow derived macrophages (n= 4 fields of view × 3 replicates, *** P<0.001, one-way ANOVA with Bonferroni’s multiple comparisons test)