Anti-MUC1 scFv h(CD28) CART, pCDCAR1 vector

Complete CAR data Funcs

Fig.1 In vitro antitumor efficacy of CAR-MUC1 T cells co-culture with OME for 6hrs.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.1 In vitro antitumor efficacy of CAR-MUC1 T cells co-culture with OME for 6hrs.

Flow cytometry tested the lysis of different tumor cells by CAR-MUC1 T cells and GFP+ T cells, respectively. CAR-MUC1 T and GFP+ T cells were coculture with tumors cell from OME for 6 h.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.2 In vitro antitumor efficacy of CAR-MUC1 T cells co-culture with HN4 for 6hrs.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.2 In vitro antitumor efficacy of CAR-MUC1 T cells co-culture with HN4 for 6hrs.

Flow cytometry tested the lysis of different tumor cells by CAR-MUC1 T cells and GFP+ T cells, respectively. CAR-MUC1 T and GFP+ T cells were coculture with tumors cell from HN4 for 6 h.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.3 In vitro antitumor efficacy of CAR-MUC1 T cells co-culture with CAL33 for 6hrs.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.3 In vitro antitumor efficacy of CAR-MUC1 T cells co-culture with CAL33 for 6hrs.

Flow cytometry tested the lysis of different tumor cells by CAR-MUC1 T cells and GFP+ T cells, respectively. CAR-MUC1 T and GFP+ T cells were coculture with tumors cell from CAL33 for 6 h.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.4 IL-2 secretion of CAR-MUC1 T cells and GFP+ T cells in coculture supernatants after different E/T ratio were measured by ELISA assay.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.4 IL-2 secretion of CAR-MUC1 T cells and GFP+ T cells in coculture supernatants after different E/T ratio were measured by ELISA assay.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.5 IFN-γ secretion of CAR-MUC1 T cells and GFP+ T cells in coculture supernatants after different E/T ratio were measured by ELISA assay.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.5 IFN-γ secretion of CAR-MUC1 T cells and GFP+ T cells in coculture supernatants after different E/T ratio were measured by ELISA assay.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.6 TNF-α secretion of CAR-MUC1 T cells and GFP+ T cells in coculture supernatants after different E/T ratio were measured by ELISA assay.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.6 TNF-α secretion of CAR-MUC1 T cells and GFP+ T cells in coculture supernatants after different E/T ratio were measured by ELISA assay.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.7 In vitro antitumor efficacy of CAR-MUC1 T cells co-culture with tumor cell after exogenous addition of IL22 recombinant protein.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.7 In vitro antitumor efficacy of CAR-MUC1 T cells co-culture with tumor cell after exogenous addition of IL22 recombinant protein.

After 72 h of exogenous addition of IL22 recombinant protein, detected the killing of CAR-MUC1 T cells against OME cell line and different HNSCC cell lines in different E/T ratios.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.8 Cytokine secretion of CAR-MUC1 T cells and GFP+ T cells in coculture supernatants with different E/T ratio after exogenous addition of IL22 addition.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.8 Cytokine secretion of CAR-MUC1 T cells and GFP+ T cells in coculture supernatants with different E/T ratio after exogenous addition of IL22 addition.

After 72 hours of exogenous addition of IL22 recombinant protein, cytokine secretion in coculture supernatants after different E/T ratio were measured by ELISA assay.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.9 Microscopic observation of the ability of two different kinds of CAR-T cells and GFP+T cells to capture cells in 1:2 E/T ratio.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.9 Microscopic observation of the ability of two different kinds of CAR-T cells and GFP+T cells to capture cells in 1:2 E/T ratio.

In CAR-MUC1 T cells group, a little number of CAR-MUC1 T cells agglomerate around tumor cells; In CAR-MUC1-IL22 T cells group, large number of CAR-MUC1-IL22 T cells agglomerate around tumor cells (Red arrow is point to the tumor cell; the blue arrow is point to the T cell or two kinds of CAR-T cells).

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.10 In vitro antitumor efficacy of CAR-MUC1 T cells co-culture with tumor cell after for 72 h.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.10 In vitro antitumor efficacy of CAR-MUC1 T cells co-culture with tumor cell after for 72 h.

Three different T cells were incubated with tumor cell (Cal33) and non-neoplastic epithelial cell (OME) at a low E/T ratio (1:1; 1:2; 1:5; 1:10) for 72 h,
flow cytometry tested the apoptosis rate of tumor cells.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.11 Representative T-cell expansion profile of three different T cells.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.11 Representative T-cell expansion profile of three different T cells.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.12 ELISA detected the secretion of IL22 in the culture supernatant.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.12 ELISA detected the secretion of IL22 in the culture supernatant.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.13 ELISA detected the secretion of IL22 in the coculture supernatant of different E/T ratio.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.13 ELISA detected the secretion of IL22 in the coculture supernatant of different E/T ratio.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.14 ELISA detected the secretion of IL-2/IFN-γ/TNF-α in the coculture supernatant of low E/T ratio.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.14 ELISA detected the secretion of IL-2/IFN-γ/TNF-α in the coculture supernatant of low E/T ratio.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.15 CAR-T cells induce tumor degradation of HNSCC in vivo.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.15 CAR-T cells induce tumor degradation of HNSCC in vivo.

1E6 fLuc+ HN4 tumor cells were injected into the subcutaneous tissue in NSG mouse. After 10 d, tail intravenous injection of CAR T cells (n = 5). The mice were sacrificed on day 40 for tumor analysis. B, Bioluminescence images of four treatment groups mice.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.16 CAR-T cells induce tumor degradation of HNSCC in vivo.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.16 CAR-T cells induce tumor degradation of HNSCC in vivo.

Total body bioluminescence units quantitate were measured by bioluminescence photometry and flux values (photons per second, P < .001).

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.17 CAR-T cells induce tumor degradation of HNSCC in vivo.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.17 CAR-T cells induce tumor degradation of HNSCC in vivo.

Tumor volume growth curves of four treatment groups (P < .001).

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.18 CAR-T cells induce tumor degradation of HNSCC in vivo.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.18 CAR-T cells induce tumor degradation of HNSCC in vivo.

T-cell infiltration statistics in four treatment groups (P < .01).

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.

Complete CAR data Funcs

Fig.19 CAR-T cells induce tumor degradation of HNSCC in vivo.

CAR Construction : SM3/HMFG2 scfv-41BB-CD3ζ-IL22

Fig.19 CAR-T cells induce tumor degradation of HNSCC in vivo.

CD3 in tumor tissue was detected using the anti-CD3 antibody (Abcam 5690, dilution 1:200); cell nuclei were stained with DAPI.

Mei, Z., Zhang, K., Lam, A. K. Y., Huang, J., Qiu, F., Qiao, B., & Zhang, Y. (2020). MUC1 as a target for CAR‐T therapy in head and neck squamous cell carinoma. Cancer medicine, 9(2), 640-652.