Extended Data Fig. 8: ho22R and hoGCSFR signalling enhance the antitumor potency of CD19 CAR-T cells and promote T cell stemness.

a, CD19 CAR, ho22R, hoGCSFR constructs were introduced via retroviral vectors. Expression levels of CD19 CAR were measured by Myc-tag fluorescence using flow cytometry. Expression levels of ho22R and hoGCSFR were measured by YFP fluorescence using flow cytometry. Shown are representative flow cytometry plots of transduction efficiency. b, pSTAT-1, -3, -4, and -5 signalling dose-response curves (n = 2 biologically independent samples). Data are presented as the MFI of pSTAT against the log₁₀ concentration of MSA-hoIL2. c,d, CD19 CAR T, ho22R CD19 CAR T, and hoGCSFR CD19 CAR T cells were cocultured with NALM6-Luciferase (NALM6-Luc) cells or Raji cells at different E:T ratios in the presence of MSA-hoIL2 (100 nM) for 24 h (n  =  3 biologically independent samples). Shown are the percentages of killing of NALM6-Luc cells (c) and Raji cells (d). e–g, CD19 CAR T, ho22R CD19 CAR T, and hoGCSFR CD19 CAR T cells were cultured in MSA-hoIL2 (100 nM) supplemented T cell medium for 3 days (n  =  3 biologically independent samples). e, Shown are MFI of CD62L and CD45RA among CD19 CAR T cells. f, Representative flow cytometry plots showing the gating strategy for CD45RA⁺ CD62L⁺ CCR7⁺ CD95⁺ subpopulations in NT, hoGCSFR, and ho22R CD3⁺ NY-ESO-1 TCR-T cells. g, Frequencies of CD45RA⁺ CD62L⁺ CCR7⁺ CD95⁺ cells. h–j, NT and oGCSFR CD8⁺ human T cells were stimulated with MSA-hoIL2 (500 nM) for 72 h, followed by flow cytometry analysis (n = 6 biologically independent samples). h, Representative flow cytometry plots of Mac-1⁺ and CD66b⁺ cells among CD8⁺ T cells. i,j, Shown are the frequencies of Mac-1⁺ (i) and CD66b⁺ (j) cells among CD8⁺ T cells. All data represent mean ± s.e.m. and are analysed by two-tailed Student’s t-test (i,j), one-way ANOVA (g), or two-way (c–e) ANOVA with Tukey’s post-test.

Source data