Fig. 7: In vivo rhIL-7-hyFc therapy enhances both the quantity and quality of CAR T cells. | Nature Communications

Fig. 7: In vivo rhIL-7-hyFc therapy enhances both the quantity and quality of CAR T cells.

From: A long-acting interleukin-7, rhIL-7-hyFc, enhances CAR T cell expansion, persistence, and anti-tumor activity

Fig. 7

a–g Balb/c mice were given cyclophosphamide 250 mg/kg on day −1, then 2 × 106 CD45.1+ UTD or mCART19 cells on day 0, followed by rhIL-7-hyFc on day +1. Spleens were harvested on days +7 and +14 after T cell injection (n = 3/group). Bar graphs represent median values, with each data point representing one mouse. a Spleen sizes (left, day +14) and cell numbers (right) were significantly increased in mice given rhIL-7-hyFc. b Fractionation of spleen cell subsets shows that the majority of cells are derived from the CD45.2+ recipient. c Absolute numbers of mCART19 cells were higher in the spleens of mice after rhIL-7-hyFc treatment. d Apoptosis was reduced in mCART19 at day +14 with rhIL-7-hyFc. e Both UTD and mCART19 show marked proliferation in response to rhIL-7-hyFc on day +7, which is better sustained at day +14 for mCART19. f, g On day +7 mCART19 was purified from spleens and subject to in vitro functional assays. f mCART19 cytotoxicity against A20 tumor cells was measured at various E:T ratios by BLI after 48 h of incubation. Each line represents mCART19 cells extracted from one mouse. g Intracellular IFNγ and TNFα production of mCART19 was measured after 4 h of incubation with A20 tumor cells. h-j NSG mice received 5×105 RamosCBR-GFP followed by UCART19 treatment and rhIL-7-hyFc on days +1 and +15. Spleens were harvested from mice on day +28. h Spleens harvested from rhIL-7-hyFc-treated mice were notably larger than controls. i, j Target cell viability was determined by FACS-based cytotoxicity assay of UCART19 from day 0 (input) as compared to UCART19 derived from spleens of rhIL-7-hyFc treated mice. The original Ramos tumor cells (i) or an independent CD19+ B cell line, Nalm6 cells (j) were incubated with CD34-purified UCART19 at E:T ratios ranging from 4:1 to 1:4 for 20 h. Bar graphs represent median values, with each data point representing a technical replicate. All p values were determined by two-sided unpaired Student’s t test. p Values and source data are provided in the Source data file. ns: not significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001.

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