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Volume 19, Issue 5, Pages 617-628 (May 2017) Dendritic cells infected by Ad-sh-SOCS1 enhance cytokine-induced killer (CIK) cell immunotherapeutic efficacy in cervical cancer models Yi Zheng, Bicheng Hu, Shenggao Xie, Xiaofan Chen, Yuqian Hu, Wanping Chen, Shanshan Li, Bo Hu Cytotherapy Volume 19, Issue 5, Pages 617-628 (May 2017) DOI: 10.1016/j.jcyt.2017.01.008 Copyright © 2017 International Society for Cellular Therapy Terms and Conditions

Figure 1 The IL-12 and IFN-γ in DC culture supernatant were measured by ELISA. Ad-sh-SOCS1 and Ad-sh-mSOCS1 promoted the expression of (A) IL-12 and (B) IFN-γ (P < 0.01). Moreover, the expression of IL-12 and IFN-γ in DCs infected by Ad-sh-SOCS1 was much higher than that by Ad-sh-mSOCS1 (P < 0.05). *P < 0.05, **P < 0.01. Cytotherapy 2017 19, 617-628DOI: (10.1016/j.jcyt.2017.01.008) Copyright © 2017 International Society for Cellular Therapy Terms and Conditions

Figure 2 The expansion fold comparison of CIKs and DC-CIKs on day 10 and day 14. Ad-sh-SOCS1-infected DCs promoted the more rapid expansion of CIKs either on day 10 or 14 compared with others. The expansion folds of different CIKs (day 14) were higher that its counterpart (day 10). Cytotherapy 2017 19, 617-628DOI: (10.1016/j.jcyt.2017.01.008) Copyright © 2017 International Society for Cellular Therapy Terms and Conditions

Figure 3 The subpopulations of CIKs treated with DCs, mgmDCs or gmDCs. The percent of CD3+CD4+ and CD4+CD25+ from gmDCs-induced CIKs was the lowest among all groups on either day 10 or 14 (P < 0.05), meanwhile the percent of CD3+CD56+ from gmDC-induced CIKs was highest among all groups at days 10 and 14 (P < 0.05). Cytotherapy 2017 19, 617-628DOI: (10.1016/j.jcyt.2017.01.008) Copyright © 2017 International Society for Cellular Therapy Terms and Conditions

Figure 4 The IL-12 and IFN-γ level from different groups were analyzed using ELISA. The level of IL-12 and IFN-γ from DCs-activated CIKs were higher than that from CIKs alone (P < 0.01). The level of IL-12 from gmDC- or mgmDC-activated CIKs were higher than that from DCs-CIKs, and the level of IL-12 from gmDC-activated CIKs were higher than that from mgmDC-activated CIKs, whereas the IFN-γ from gmDC-activated CIKs were higher than that from DCs-CIKs or mgmDCs-CIKs (P < 0.05). Cytotherapy 2017 19, 617-628DOI: (10.1016/j.jcyt.2017.01.008) Copyright © 2017 International Society for Cellular Therapy Terms and Conditions

Figure 5 The cytotoxicity of different DC-activated CIKs on target cells. The cytotoxicity of DC-activated CIKs on TC-1 cells was higher than that of CIKs alone (P < 0.05) at various E:T ratios. The cytotoxicity of gmDC CIKs on TC-1 or HeLa cells was higher than that from other groups. The cytotoxicity of DCs-CIKs on days 10 and 14 showed no significant difference (P > 0.05) at various E:T ratios. Cytotherapy 2017 19, 617-628DOI: (10.1016/j.jcyt.2017.01.008) Copyright © 2017 International Society for Cellular Therapy Terms and Conditions

Figure 6 The survival time and rate of mice treated with different DCs-activated CIKs. The survival time of mice treated with gmDC-activated CIKs (day 10 or 14) was longer than the other groups (P < 0.01). The survival time of mice treated by different type of CIKs was longer than that from PBS-treated group. The survival time of mice treated by different DCs-activated CIKs was longer than that of treated with CIKs alone (day 10 or 14). The survival time of mice treated with DCs-CIKs (day 10 and 14) and mgmDC-CIKs (day 10 and 14) had no significant difference (P > 0.05). Cytotherapy 2017 19, 617-628DOI: (10.1016/j.jcyt.2017.01.008) Copyright © 2017 International Society for Cellular Therapy Terms and Conditions

Figure 7 The serum IL-12 and IFN-γ levels of treated mice. The IL-12 levels of mice treated with gmDC-CIKs (days 10 and 14) were much higher than other groups (P < 0.05), and the IL-12 level in gmDC-CIKs (day 10) group was much higher than gmDC-CIKs (day 14) group (P < 0.05). IL-12 level in serum from these groups treated by DC in combination with CIKs was higher than that treated by CIKs alone, but no significant difference among these groups except gmDC-CIKs. In addition, the IFN-γ level in serum from mice treated by gmDC-CIKs (either day 10 or day 14) was higher than that of others (P < 0.05), and no significant difference among other groups (P > 0.05). Cytotherapy 2017 19, 617-628DOI: (10.1016/j.jcyt.2017.01.008) Copyright © 2017 International Society for Cellular Therapy Terms and Conditions

Figure 8 Hematoxylin and eosin staining of transplantation tumors in nude mice (10 × 10). (A) PBS; (B) CIK (day 10); (C) CIK (day 14); (D) DC-CIK (day 10); (E) DC-CIK (day 14); (F) mgmDC-CIK (day 10); (G) mgmDC-CIK (day 14); (H) gmDC (day 10); (I) gmDC-CIK (day 14). Cytotherapy 2017 19, 617-628DOI: (10.1016/j.jcyt.2017.01.008) Copyright © 2017 International Society for Cellular Therapy Terms and Conditions