title: Induction of tumor-specific cytotoxicity in tumor infiltrating lymphocytes by HPV16 and HPV18 E7-pulsed autologous dendritic cells in patients with cancer of the uterine cervix.
abstract: OBJECTIVE
  To evaluate the potential of autologous dendritic cells (DC) pulsed with HPV16 and HPV18 E7 oncoprotein in restoring tumor-specific cytotoxicity in populations of tumor infiltrating lymphocytes (TIL) for adoptive immunotherapy of cervical cancer patients.
  
  
  METHODS
  Full-length E7-pulsed DC-stimulated CD8(+) T cells derived from peripheral blood (PBL) and from tumor tissues (TIL) were tested and compared for their ability to induce a HLA class-I-restricted cytotoxic T lymphocyte (CTL) response against autologous tumor cells. In addition, in order to correlate cytotoxic activity by CTL with a particular lymphoid subset, analysis of surface antigens and intracellular CD3 zeta chain and two-color flow cytometric analysis of intracellular cytokine expression (IFN-gamma vs IL-4) at the single cell level were performed.
  
  
  RESULTS
  DC stimulation induced powerful cytotoxicity against autologous tumor target cells by TIL-derived CD8(+) T cells from all three cervical cancer patients, while autologous Epstein-Barr virus-transformed lymphoblastoid cell lines were not lysed. Killing of autologous tumor cells was higher by CD8(+) T cells from TIL compared to PBL (P > 0.01) and was more strongly inhibited by anti-HLA class I MAb (P > 0.05). Phenotypically, all CTL populations were CD3(+)/CD8(+), with higher levels of CD56 expression by TIL-derived CTL. Finally, although a marked Type 1 cytokine bias (i.e., IFN-gamma(high)/IL-4(low)) was observable in both PBL- and TIL-derived DC-stimulated CD8(+) T cell populations, TIL-derived CD8(+) T cells showed a higher percentage of IFN-gamma-positive cells compared to PBL.
  
  
  CONCLUSIONS
  Full-length E7-pulsed DC can consistently restore strong CD8(+) CTL responses against autologous HPV16- and HPV18-infected cervical cancer cells. DC-stimulated TIL may represent a superior source of tumor-specific CTL compared to PBL for adoptive T cell immunotherapy of patients harboring metastatic or recurrent cervical cancer refractory to standard treatment modalities.
authors.affiliation: Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, University of Arkansas for Medical Sciences, Little Rock 72205-7199, USA. santinalessandrod@uams.edu
authors.forename: Alessandro D
authors.initials: AD
authors.lastname: Santin
authors.forename: Stefania
authors.initials: S
authors.lastname: Bellone
authors.forename: Michela
authors.initials: M
authors.lastname: Palmieri
authors.forename: Barbara
authors.initials: B
authors.lastname: Bossini
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Roman
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Cannon
authors.forename: Eliana
authors.initials: E
authors.lastname: Bignotti
authors.forename: Stefania
authors.initials: S
authors.lastname: Canè
authors.forename: Sergio
authors.initials: S
authors.lastname: Pecorelli
mesh_terms: D000328:Adult; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D004268:DNA-Binding Proteins; D003713:Dendritic Cells / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007371:Interferon-gamma / Q000096:biosynthesis; D015847:Interleukin-4 / Q000096:biosynthesis; D007962:Leukocytes / Q000276:immunology / Q000378:metabolism; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology* / Q000378:metabolism; D008565:Membrane Proteins / Q000096:biosynthesis; D009856:Oncogene Proteins, Viral / Q000276:immunology*; D050725:Papillomavirus E7 Proteins; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism; D002583:Uterine Cervical Neoplasms / Q000276:immunology* / Q000628:therapy*
chemical_list: D004268:DNA-Binding Proteins; C052633:E7 protein, Human papillomavirus type 18; D008565:Membrane Proteins; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D011948:Receptors, Antigen, T-Cell; C054979:antigen T cell receptor, zeta chain; C059731:oncogene protein E7, Human papillomavirus type 16; D015847:Interleukin-4; D007371:Interferon-gamma