title: Therapeutic dendritic cell vaccine preparation using tumor RNA transfection: a promising approach for the treatment of prostate cancer.
abstract: BACKGROUND
  Early prostate adenocarcinoma can be diagnosed through seric prostate-specific antigen (PSA) screenings. However, a fraction of patients progress to an incurable metastatic disease. Therefore, novel therapies for treating these patients are extremely desirable. Therapeutic vaccines based on Dendritic Cells (DCs) carrying tumor antigens have emerged as a promising strategy to initiate an immune response against tumor cells. These vaccines can be prepared using different methodologies, such as the application of tumor mRNA described in this work.
  
  
  METHODS
  Mature and immature DCs were obtained in vitro by adding specific cytokines to monocyte cell cultures. RNA extracted from prostate tumor lineage (LNCAP) was introduced into these cells by electroporation and co-incubation. Transfection success was measured by immunocytochemistry of the PSA expression level in DCs.
  
  
  RESULTS
  Cell surface markers, including CD14, CD80, CD86, CCR7, CD11c, and CD1a, confirmed mature and immature DC phenotypes. Both cell maturation stages were successfully used for RNA introduction as shown by PSA characterization.
  
  
  CONCLUSIONS
  Our data support the use of mature and immature DCs for vaccine preparation with either RNA electroporation or RNA co-incubation. The highest efficiency, however, was observed when RNA was delivered by electroporation into mature DCs. Due to in vitro RNA transcription, this method allows small tumors to be used for DC vaccine preparation; it is therefore a promising approach for the treatment of metastatic prostate cancer.
authors.affiliation: Oncocell Division, Genoa Biotechnology SA, Alameda Ministro Rocha Azevedo, 346, 1st floor, 01410-000, São Paulo, SP, Brazil. jmscanavez@genoabiotec.com.br
authors.forename: Juliana M
authors.initials: JM
authors.lastname: Sousa-Canavez
authors.forename: Flavio C
authors.initials: FC
authors.lastname: Canavez
authors.forename: Kátia R M
authors.initials: KR
authors.lastname: Leite
authors.forename: Luiz H
authors.initials: LH
authors.lastname: Camara-Lopes