Page 248 - Haematologica Vol. 109 - July 2024

ARTICLE - Application of CAAR T-cell therapy in ITP J. Zhou et al. A
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Figure 3. GPIbα chimeric autoantibody receptor expression on primary human T cells directs specific cytolysis of anti-GPIbα B cells in vitro. (A) Hybridoma cells (Gvb1/Gvb2/Gvb3/Gvb4) were co-incubated with non-transduced T cells (NTD T) or GPIbα chi- meric autoantibody receptor (CAAR) T cells at the indicated effector to target (E:T) ratios for 16 hours. Specific lysis was measured by lactate dehydrogenase (LDH) release into the supernatant. (B-E) NTD T or LBD-mut223k T cells were co-incubated with con- trol hybridoma (PE) and anti-GPIbα hybridomas (APC) at an E:T ratio of 5:1. The ratio of the target hybridoma to total control hybridoma cells was 3:1. Cytotoxicity was evaluated at 24 hours using flow cytometry, as indicated by the changes in the ratio of the target hybridoma to the control hybridoma. **P<0.01; ****P<0.0001. (F) Cytokines (interleukin [IL]-2, interferon [IFN]-γ) pro- duced by NTD T or GPIbα CAAR T cells after 5:1 co-culture with each anti-GPIbα hybridoma cell for 16 hours, as measured in the supernatant by enzyme-linked immunosorbant assay.
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