Journal: Journal for Immunotherapy of Cancer

Article Title: Delivery of CD47 blocker SIRPα-Fc by CAR-T cells enhances antitumor efficacy

doi: 10.1136/jitc-2021-003737

Figure Lengend Snippet: In vitro antitumor function of CAR-T cells. (A, B) The cytotoxic effect of CAR-T cells with different trop2-positive murine tumor cells. The expression of IFN-γ in CD3 + T cells when cocultured with MC38-Trop2 + cells (C), and 4T1-Trop2 + cells (D) were conducted by flow cytometry. For the in vitro phagocytosis assay, BMDMs were cocultured with CFSE-labeled MC38-Trop + cells at a 1:4 ratio in the supernatant of different T cells (E) or the medium of cocultured T cells with tumor cells (F). The cells were harvested after 4 hours coculture, and primary macrophages were identified by flow cytometry using anti-F4/80. Phagocytosis rate was determined as the percentage of CFSE + macrophages. (mean±SEM, n=3). Representative data from at least three independent experiments are shown. *p<0.05, **p<0.01, ****p<0.0001. BMDMs, bone marrow-derived macrophages; CAR, chimeric antigen receptor; CFSE, carboxyfluorescein diacetate succinimidyl ester; ns, not significant; UTD, untransduced.

Article Snippet: Mouse T cells isolated from wild-type C57BL/6 or BALB/c female mouse splenocytes were purified with a mouse CD3 + T cell negative enrichment kit (BD, USA).

Techniques: In Vitro, Expressing, Flow Cytometry, Phagocytosis Assay, Labeling, Derivative Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Delivery of CD47 blocker SIRPα-Fc by CAR-T cells enhances antitumor efficacy

doi: 10.1136/jitc-2021-003737

Figure Lengend Snippet: Secretion of SIRPα-Fc by CAR T cells modulate the immune cells in tumor-bearing mouse. Flow cytometry was used to detect the proportion of CAR + in tumor-bearing mouse spleen. (A) CAR positive rate in CD3 + T cells. (B) CAR positive rate in CD4 + T cells, (C) CAR positive rate in CD8 + T cells. (D) CD44 and CD62L of CD3 + CAR + T cells in mouse spleen. Flow cytometry analysis of (E) CD4 + CAR + T cells, (F) CD8 + CAR + T cells, (G) CD45 + CD11c + cells, and (H) CD11b + Ly6G + cells in tumor tissues. (I) Frequency of M1-like (CD206 — MHCII high ) and (J) M2-like (CD206 + MHC II low/neg ) TAMs cell populations in CD45 + CD3 — CD11b + F4/80 + in tumor tissues. All data are presented as mean±SEM from experiments, N=3 mice/group. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. CAR, chimeric antigen receptor; MHC, major histocompatibility complex; ns, not significant; SIRPα, signal regulatory protein α; TAM, tumor-associated macrophages; UTD, untransduced.

Article Snippet: Mouse T cells isolated from wild-type C57BL/6 or BALB/c female mouse splenocytes were purified with a mouse CD3 + T cell negative enrichment kit (BD, USA).

Techniques: Flow Cytometry