title: Colonization of xenograft tumors by oncolytic vaccinia virus (VACV) results in enhanced tumor killing due to the involvement of myeloid cells.
abstract: The mechanisms by which vaccinia virus (VACV) interacts with the innate immune components are complex and involve different mechanisms. iNOS-mediated NO production by myeloid cells is one of the central antiviral mechanisms and this study aims to investigate specifically whether iNOS-mediated NO production by myeloid cells, is involved in tumor eradication following the virus treatment.
  
  
  
  Human colon adenocarcinoma (HCT-116) xenograft tumors were infected by VACV. Infiltration of iNOS+myeloid cell population into the tumor, and virus titer was monitored following the treatment. Single-cell suspensions were stained for qualitative and quantitative flow analysis. The effect of different myeloid cell subsets on tumor growth and colonization were investigated by depletion studies. Finally, in vitro culture experiments were carried out to study NO production and tumor cell killing. Student's t test was used for comparison between groups in all of the experiments.
  
  
  
  Infection of human colon adenocarcinoma (HCT-116) xenograft tumors by VACV has led to recruitment of many CD11b+ly6G+myeloid-derived suppressor cells (MDSCs), with enhanced iNOS expression in the tumors, and to an increased intratumoral virus titer between days 7 and 10 post-VACV therapy. In parallel, both single and multiple rounds of iNOS-producing cell depletions caused very rapid tumor growth within the same period after virus injection, indicating that VACV-induced iNOS+MDSCs could be an important antitumor effector component. A continuous blockade of iNOS by its specific inhibitor, L-NIL, showed similar tumor growth enhancement 7-10 days post-infection. Finally, spleen-derived iNOS+MDSCs isolated from virus-injected tumor bearing mice produced higher amounts of NO and effectively killed HCT-116 cells in in vitro transwell experiments.
  
  
  
  We initially hypothesized that NO could be one of the factors that limits active spreading of the virus in the cancerous tissue. In contrast to our initial hypothesis, we observed that PMN-MDSCs were the main producer of NO through iNOS and NO provided a beneficial antitumor effect, The results strongly support an important novel role for VACV infection in the tumor microenvironment. VACV convert tumor-promoting MDSCs into tumor-killing cells by inducing higher NO production.
authors.affiliation: Department of Biochemistry, Biocenter, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.|San Diego Science Center, Genelux Corporation, San Diego, CA, USA.
authors.forename: Mehmet Okyay
authors.initials: MO
authors.lastname: Kilinc
authors.affiliation: Department of Biochemistry, Biocenter, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.
authors.forename: Klaas
authors.initials: K
authors.lastname: Ehrig
authors.affiliation: San Diego Science Center, Genelux Corporation, San Diego, CA, USA.
authors.forename: Maysam
authors.initials: M
authors.lastname: Pessian
authors.affiliation: San Diego Science Center, Genelux Corporation, San Diego, CA, USA.|Department of Radiation Medicine and Applied Sciences, Rebecca & John Moores Comprehensive Cancer Center, University of California, San Diego, CA, USA.
authors.forename: Boris R
authors.initials: BR
authors.lastname: Minev
authors.affiliation: Department of Biochemistry, Biocenter, University of Würzburg, Am Hubland, 97074, Würzburg, Germany. aladar.szalay@virchow.uni-wuerzburg.de.|San Diego Science Center, Genelux Corporation, San Diego, CA, USA. aladar.szalay@virchow.uni-wuerzburg.de.|Department of Radiation Medicine and Applied Sciences, Rebecca & John Moores Comprehensive Cancer Center, University of California, San Diego, CA, USA. aladar.szalay@virchow.uni-wuerzburg.de.
authors.forename: Aladar A
authors.initials: AA
authors.lastname: Szalay
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D003602:Cytotoxicity, Immunologic; D045325:HCT116 Cells; D006801:Humans; D007700:Kinetics; D008297:Male; D008819:Mice, Nude; D022423:Myeloid Cells / Q000276:immunology*; D009504:Neutrophils / Q000473:pathology; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D050504:Oncolytic Viruses / Q000502:physiology*; D047368:Tumor Burden; D014616:Vaccinia virus / Q000502:physiology*; D023041:Xenograft Model Antitumor Assays
chemical_list: D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II
keywords: Antitumor immune response; Antiviral immunity; Innate immune system; MDSCs; NO; Oncolytic virus therapy; VACV; iNOS