Cytokine-induced killer (CIK) cells bound with anti-CD3/anti-CD133 bispecific antibodies target CD133high cancer stem cells in vitro and in vivo

•We generate anti-CD3/anti-CD133 BsAb to target CD133high cancer cells.•BsAb-CIKs kill CD133high cancer cells significantly more than CIKs in vitro.•BsAb-CIKs inhibit CD133high tumor growth significantly more than CIKs in vivo.•CD133 targeting modulates S100P, IL-18 bp and STAT1 expression in tumor cells.•We report a novel immunotherapy for cancer containing CD133high cancer stem cells.

CD133 is a common marker of cancer stem cells (CSCs). We generated an anti-CD3/anti-CD133 bispecific antibody (BsAb) and bound it to the cytokine-induced killer (CIK) cells as effector cells (BsAb-CIK) to target CD133high CSCs. The killing of CD133high pancreatic (SW1990) and hepatic (Hep3B) cancer cells by the BsAb-CIK cells was significantly (p < 0.05) higher than the killing by the parental CIK or by CIK cells bound with anti-CD3 (CD3-CIK) without CD133 targeting. In nude mice, the BsAb-CIK cells inhibited CD133high tumor growth significantly (p < 0.05) more than that by CIK or CD3-CIK cells, or by the BsAb alone. BsAb-CIK cells co-cultured with CD133high cells produced significantly (p < 0.05) higher amount of IFN-γ. Treatment with the BsAb-CIK cells significantly downregulated the expression of S100P and IL-18 bp, but upregulated STAT1. The findings may help with the development of novel immunotherapies for patients with cancer containing CD133high CSCs by selectively targeting this cell population.