title: Co-introduced functional CCR2 potentiates in vivo anti-lung cancer functionality mediated by T cells double gene-modified to express WT1-specific T-cell receptor.
abstract: OBJECTIVE
  Although gene-modification of T cells to express tumor-related antigen-specific T-cell receptor (TCR) or chimeric antigen receptor (CAR) has clinically proved promise, there still remains room to improve the clinical efficacy of re-directed T-cell based antitumor adoptive therapy. In order to achieve more objective clinical responses using ex vivo-expanded tumor-responsive T cells, the infused T cells need to show adequate localized infiltration into the tumor.
  
  
  RESULTS
  Human lung cancer cells variously express a tumor antigen, Wilms' Tumor gene product 1 (WT1), and an inflammatory chemokine, CCL2. However, CCR2, the relevant receptor for CCL2, is rarely expressed on activated T-lymphocytes. A HLA-A2402(+) human lung cancer cell line, LK79, which expresses high amounts of both CCL2 and WT1 mRNA, was employed as a target. Normal CD8(+) T cells were retrovirally gene-modified to express both CCR2 and HLA-A*2402-restricted and WT1(235-243) nonapeptide-specific TCR as an effector. Anti-tumor functionality mediated by these effector cells against LK79 cells was assessed both in vitro and in vivo. Finally the impact of CCL2 on WT1 epitope-responsive TCR signaling mediated by the effector cells was studied. Introduced CCR2 was functionally validated using gene-modified Jurkat cells and human CD3(+) T cells both in vitro and in vivo. Double gene-modified CD3(+) T cells successfully demonstrated both CCL2-tropic tumor trafficking and cytocidal reactivity against LK79 cells in vitro and in vivo. CCL2 augmented the WT1 epitope-responsive TCR signaling shown by relevant luciferase production in double gene-modified Jurkat/MA cells to express luciferase and WT1-specific TCR, and CCL2 also dose-dependently augmented WT1 epitope-responsive IFN-γ production and CD107a expression mediated by these double gene-modified CD3(+) T cells.
  
  
  CONCLUSIONS
  Introduction of the CCL2/CCR2 axis successfully potentiated in vivo anti-lung cancer reactivity mediated by CD8(+) T cells double gene-modified to express WT1-specific TCR and CCR2 not only via CCL2-tropic tumor trafficking, but also CCL2-enhanced WT1-responsiveness.
authors: Department of Bioregulatory Medicine, Ehime University Graduate School of Medicine, Ehime, Japan.
authors: Hiroaki
authors: H
authors: Asai
authors: Hiroshi
authors: H
authors: Fujiwara
authors: Jun
authors: J
authors: An
authors: Toshiki
authors: T
authors: Ochi
authors: Yukihiro
authors: Y
authors: Miyazaki
authors: Kozo
authors: K
authors: Nagai
authors: Sachiko
authors: S
authors: Okamoto
authors: Junichi
authors: J
authors: Mineno
authors: Kiyotaka
authors: K
authors: Kuzushima
authors: Hiroshi
authors: H
authors: Shiku
authors: Hirofumi
authors: H
authors: Inoue
authors: Masaki
authors: M
authors: Yasukawa
mesh_terms: D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D008175:Lung Neoplasms / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D051379:Mice; D016853:Muromonab-CD3 / Q000494:pharmacology; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D054390:Receptors, CCR2 / Q000235:genetics* / Q000378:metabolism; D015203:Reproducibility of Results; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D014162:Transfection; D014183:Transplantation, Heterologous; D025721:WT1 Proteins / Q000276:immunology*
chemical_list: D007376:Interleukin-2; D016853:Muromonab-CD3; D011948:Receptors, Antigen, T-Cell; D054390:Receptors, CCR2; D025721:WT1 Proteins