title: Membrane-bound complement regulatory proteins inhibit complement activation by an immunotherapeutic mAb in a syngeneic rat colorectal cancer model.
abstract: MAb-mediated immunotherapy offers a potential tool for destroying metastasizing colorectal tumor cells. Promising results have been obtained by using xenograft models. However, overexpression of membrane-bound complement regulatory proteins (mCRP) impedes complement-mediated destruction of tumor cells in vitro. mCRP operate in a species selective manner. Therefore a syngeneic animal model is needed to investigate the contribution of mCRP in mAb-mediated immunotherapy. Here we present a syngeneic rat colorectal carcinoma model, which fulfills the conditions necessary to investigate the effect of mCRP expression on mAb-mediated immunotherapy of metastases of solid tumors.CC531 rat colorectal cancer cells were injected subcapsularly into the liver of syngeneic WAG/Rij rats. Four mAb (MG1(IgG2a), MG2(IgG2a), MG3(IgG3) and MG4(2a)(IgG2a)) directed against CC531 cells, were tested for their complement activating abilities in vitro and tumor homing capacities in vivo. Only MG4(2a) was found to activate complement in vitro and home to the tumor cells in vivo. This mAb induced C3-deposition and C-mediated lysis of CC531 cells in vitro when the effects of the C-inhibitors Crry/p65 and CD59 were neutralized. This implies an important role for these mCRP in restricting the effector functions of tumor-associated mAb on these cells. Although C activation could be induced by MG4(2a) in situ on tumor tissue sections, no deposition of C3 could be found on the tumor cells positive for MG4(2a) in vivo. This suggests that complement activation in vivo was inhibited by mCRP. The results indicate the suitability of this syngeneic animal model for studying the effects of mAb immunotherapy. However, the effect of mCRP on tumor cells need to be overcome, e.g. by the use of mAb against tumor antigens and mCRP.
authors.affiliation: Department of Pathology, Leiden University Medical Center, L1-Q, Albinusdreef 2, P.O. Box 9600, 2300 RC Leiden, The Netherlands. k.gelderman@lumc.nl
authors.forename: K A
authors.initials: KA
authors.lastname: Gelderman
authors.forename: J
authors.initials: J
authors.lastname: Hakulinen
authors.forename: M
authors.initials: M
authors.lastname: Hagenaars
authors.forename: P J K
authors.initials: PJ
authors.lastname: Kuppen
authors.forename: S
authors.initials: S
authors.lastname: Meri
authors.forename: A
authors.initials: A
authors.lastname: Gorter
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000627:therapeutic use*; D000954:Antigens, Surface; D018958:CD55 Antigens / Q000032:analysis / Q000502:physiology*; D018957:CD59 Antigens / Q000032:analysis / Q000502:physiology*; D015179:Colorectal Neoplasms / Q000276:immunology / Q000628:therapy*; D003167:Complement Activation; D003165:Complement System Proteins / Q000276:immunology; D051381:Rats; D011956:Receptors, Cell Surface; D011951:Receptors, Complement / Q000032:analysis / Q000502:physiology*; D014407:Tumor Cells, Cultured
chemical_list: D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D018958:CD55 Antigens; D018957:CD59 Antigens; C513105:Cr1l protein, rat; D011956:Receptors, Cell Surface; D011951:Receptors, Complement; D003165:Complement System Proteins