title: Development of inCVAX,In situCancer Vaccine, and Its Immune Response in Mice with Hepatocellular Cancer.
abstract: Manipulation of immune system toward the rejection of established cancers has become the standard of care in some patients. Here we propose the development of anin situautologous cancer vaccine, inCVAX, for the treatment of hepatocellular cancer (HCC). inCVAX is based on the induction of local immunogenic cancer cell death combined with local dendritic cell stimulation by intratumoral injection of the immune-activatorN-dihydro-galacto-chitosan (GC). In a first set of experiments, cellular and molecular studies were performed to investigate the effect of inCVAX on immune activation in a murine model of HCC that we previously developed. Once large tumors were formed in mice, the tumor is surgically exposed and a laser fiber was inserted into the center of an individual tumor mass. Using a 10 mm diffuser tip, laser irradiation of 1.5 W was applied to heat the tumor at different durations (6-10 min) to assess tolerability of photothermal application at different temperatures. The laser application was followed by immediate injection of GC, and each mouse received one laser treatment and one GC injection. ELISA was used to assess the level of cytokines; immunohistochemical staining was conducted to analyze the effect of inCVAX on immune cell tumor-filtration and expression of tumor-specific antigens (TSAs) and tumor-associated antigens (TAAs). Results indicate that survival correlated to thermal exposure. At lower temperatures the photothermal effect was sufficient to induce tumor necrosis, but without obvious complication to the mice, although at these temperatures the treatment didn't alter the level of TSAs and TAAs, so further optimization is suggested. Nevertheless, in response to the inCVAX treatment, cytotoxic cytokine IFN-γ was significantly increased, but suppressive cytokine TGF-β was dramatically reduced. Furthermore, inCVAX prompted tumor infiltration of CD3+, CD4+, and CD8+T cells; but modulated macrophage subsets differently. In conclusion, while the protocol needs further optimization, it would appear that inCVAX for the treatment of HCC activates an immune response in tumor-bearing mice, which in turn may have potential for the treatment of HCC.
authors.affiliation: Department of Surgery, University of Missouri, Columbia, MO 65212, USA ; Ellis Fischel Cancer Center, University of Missouri, Columbia, MO 65212, USA.
authors.forename: Xiaoqiang
authors.initials: X
authors.lastname: Qi
authors.affiliation: Immunophotonics Inc., 4320 Forest Park Avenue #303, St. Louis, Missouri 63108, USA.
authors.forename: Samuel Sk
authors.initials: SS
authors.lastname: Lam
authors.affiliation: Department of Surgery, University of Missouri, Columbia, MO 65212, USA ; Ellis Fischel Cancer Center, University of Missouri, Columbia, MO 65212, USA.
authors.forename: Dai
authors.initials: D
authors.lastname: Liu
authors.affiliation: Veterinary Medical Diagnostic Laboratory, College of Veterinary Medicine, University of Missouri, Columbia, MO 65212, USA.
authors.forename: Dae Young
authors.initials: DY
authors.lastname: Kim
authors.affiliation: Department of Radiology, University of Missouri, Columbia, MO 65212; Harry S. Truman Memorial VA Hospital Biomolecular Imaging Center, USA.
authors.forename: Lixin
authors.initials: L
authors.lastname: Ma
authors.affiliation: Immunophotonics Inc., 4320 Forest Park Avenue #303, St. Louis, Missouri 63108, USA.
authors.forename: Lu
authors.initials: L
authors.lastname: Alleruzzo
authors.affiliation: Veterinary Medical and Surgery, College of Veterinary Medicine, University of Missouri, Columbia, MO 65212, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.affiliation: Immunophotonics Inc., 4320 Forest Park Avenue #303, St. Louis, Missouri 63108, USA.
authors.forename: Tomas
authors.initials: T
authors.lastname: Hode
authors.affiliation: Department of Molecular Microbiology and Immunology, University of Missouri, Columbia, MO 65212, USA.
authors.forename: Carolyn J
authors.initials: CJ
authors.lastname: Henry
authors.affiliation: Department of Surgery, University of Missouri, Columbia, MO 65212, USA ; Ellis Fischel Cancer Center, University of Missouri, Columbia, MO 65212, USA.
authors.forename: Jussuf
authors.initials: J
authors.lastname: Kaifi
authors.affiliation: Department of Surgery, University of Missouri, Columbia, MO 65212, USA ; Ellis Fischel Cancer Center, University of Missouri, Columbia, MO 65212, USA.
authors.forename: Eric T
authors.initials: ET
authors.lastname: Kimchi
authors.affiliation: Department of Surgery, University of Missouri, Columbia, MO 65212, USA ; Ellis Fischel Cancer Center, University of Missouri, Columbia, MO 65212, USA.
authors.forename: Guangfu
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of Surgery, University of Missouri, Columbia, MO 65212, USA ; Ellis Fischel Cancer Center, University of Missouri, Columbia, MO 65212, USA ; Department of Molecular Microbiology and Immunology, University of Missouri, Columbia, MO 65212, USA.
authors.forename: Kevin F
authors.initials: KF
authors.lastname: Staveley-O'Carroll
keywords: Cytokine; Hepatocellular cancer (HCC); Laser immunotherapy (LIT); Macrophage; T cell; in situ cancer vaccine (inCVAX)