Skip to main content
Fig. 4 | Journal of Hematology & Oncology

Fig. 4

From: Anti-GD2/4-1BB chimeric antigen receptor T cell therapy for the treatment of Chinese melanoma patients

Fig. 4

Functional analysis of GD2/CAR-T cells in vitro. a Cytotoxic activity of GD2/CAR-T cells. We used an LDH release assay to evaluate the cytotoxic activity of GD2.BBζ CAR-T cells and non-transduced T cells. Target cells were melanoma lines with varying GD2 expression levels. The figure illustrates the mean and SD of LDH release from 9 T cell lines after 4 h of incubation. A significant difference was detected in GD2-specific antitumor activity at each E:T ratio between GD2.BBζ CAR-T cells and control non-transduced CAR-T cells. In contrast, GD2.BBζ CAR-T cells had little antitumor activity against the GD2-negative tumor cell lines 293T, A875 (3.1% GD2 expression), and MMYC-7 (10.2% GD2 expression). b Th1 cytokine release of GD2/CAR-T cells. Non-transduced T cells and GD2.BBζ CAR-T cells were co-cultured (ratio of T lymphocytes:tumor cells of 20:1) with four different cell lines that were GD2-negative (293T) or were 27.4% GD2-positive (GAK), were 47.3% GD2-positive (HMV-II) or exhibited high (WM-266-4) levels of GD2-positive cells. Culture supernatant was collected 24 h later, and the production of IL-2, TNF-α, and IFN-γ were measured using a CBA assay. A substantial amount of IL2, TNF-α, and IFN-γ was released by GD2.BBζ CAR-T cells, and their releases were associated with the level of GD2 expression in the melanoma cells. In contrast, the release of IL2, TNF-α, and IFN-γ remained unchanged in non-transduced T cells or 293 T cells. The results are presented as the mean and SD from experiments that were performed in triplicate

Back to article page