Cotransfection of DC with TLR4 and MART-1 RNA induces MART-1-specific responses

Zeinab Abdel-Wahab, Robin Cisco, Jens Dannull, Tomio Ueno, Omar Abdel-Wahab, Matthew F. Kalady, Mark W. Onaitis, Douglas S. Tyler, Scott K. Pruitt

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

Background. Cotransfection of dendritic cells (DC) with MART-1 and constitutively active TLR4 (caTLR4) RNA enhances the maturation of DC. Materials and methods. Immature DC were cotransfected with RNA constructs encoding MART-1 and caTLR4, and CTL responses were analyzed. Results. Cotransfection of DC with MART-1 + caTLR4 enhanced the expression of CD80 and CD83 surface markers and increased the secretion of cytokines IL-6, IL-12, and TNFα. Neither the native nor the A27L-modified MART-1 RNA could induce significant DC maturation or cytokine secretion. More importantly, DC cotransfected with caTLR4 + MART-1 RNA induced MART-1-specific CTL responses of a higher magnitude than DC transfected with either the native or A27L MART-1 RNA. When the MART-1 RNA-transfected DC were treated with DC-maturing cytokines, the induced CTL were less frequent and less lytic than those induced with MART-1 + caTLR4. A 2- to 100-fold increase in MART-1 tetramer+ cells and 2- to 10-fold increases in IFNγ secretion and cytotoxicity were seen in CTL induced with MART-1 + caTLR4 compared to CTL induced with either MART-1 or A27L RNA. CTL induced with the mixed RNA displayed high percentages of CD8+ cells coexpressing CD45RA, CD56, and 2B4 antigens. Transfection with caTLR4 alone induced DC maturation, but did not induce lytic CTL, suggesting that CTL responses were induced solely by MART-1 epitopes. Conclusions. caTLR4 increases the CTL-inducing capacity of DC generating a lytic response specific for the accompanying antigen. These results demonstrate the possibility of enhancing the immunogenicity of the native MART-1 and other RNA derived from weakly immunogenic tumors in DC-based immunotherapy.

Original languageEnglish (US)
Pages (from-to)264-273
Number of pages10
JournalJournal of Surgical Research
Volume124
Issue number2
DOIs
StatePublished - Apr 2005
Externally publishedYes

Keywords

  • A27L RNA
  • CTL
  • DC
  • MART-1
  • Melanoma
  • TLR4

ASJC Scopus subject areas

  • Surgery

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  • Cite this

    Abdel-Wahab, Z., Cisco, R., Dannull, J., Ueno, T., Abdel-Wahab, O., Kalady, M. F., Onaitis, M. W., Tyler, D. S., & Pruitt, S. K. (2005). Cotransfection of DC with TLR4 and MART-1 RNA induces MART-1-specific responses. Journal of Surgical Research, 124(2), 264-273. https://doi.org/10.1016/j.jss.2004.10.002