Induction of anti-melanoma CTL response using DC transfected with mutated mRNA encoding full-length Melan-A/MART-1 antigen with an A27L amino acid substitution. Academic Article uri icon

Overview

MeSH

  • Amino Acid Substitution
  • Antigens, Neoplasm
  • Cell Line, Tumor
  • Humans
  • Immunophenotyping
  • Interferon-gamma
  • MART-1 Antigen
  • Mutation
  • Transfection

MeSH Major

  • Dendritic Cells
  • Melanoma
  • Neoplasm Proteins
  • RNA, Messenger
  • T-Lymphocytes, Cytotoxic

abstract

  • Modification of the parental immunodominant Melan-A/MART-1 peptide (MART-1(26-35)) by replacing the alanine with leucine (A27L) enhances its immunogenicity. Because of the reported advantages of RNA over peptides in DC vaccines, we sought to mutate the MART-1 gene to encode a full-length MART-1 antigen with an A27L amino acid substitution. Human DC were transfected with A27L-mutated MART-1 RNA (A27L RNA) or native MART-1 RNA, and then used to stimulate autologous T cells from a series of 8 HLA-A2+ volunteers. After three stimulations, all CTL induced with DC/A27L RNA exhibited more tetramer+ cells, and demonstrated stronger antigen-specific IFNgamma-secreting activity compared to CTL induced with DC/native RNA. A potent MART-1-specific, and predominantly class-I-restricted lysis was detected in most CTL induced with DC/A27L RNA, while native RNA-induced CTL showed minimal and non-specific lysis. HLA-A2+ DC and MART-1 negative/A2+ melanoma cells transfected with the A27L RNA were recognized and killed by MART-1-specific CTL, suggesting that these APC efficiently processed the A27L RNA and presented correct MART-1-specific epitope(s). In summary, introducing an A27L mutation into the MART-1 full-length mRNA sequence enhanced the immunogenicity of the encoded MART-1 Ag. The ease with which such a mutation can be made in RNA presents another potential advantage of using RNA for immunotherapy. Our results support considering this strategy for enhancing the immunogenicity of DC-based RNA vaccines.

publication date

  • August 2003

has subject area

  • Amino Acid Substitution
  • Antigens, Neoplasm
  • Cell Line, Tumor
  • Dendritic Cells
  • Humans
  • Immunophenotyping
  • Interferon-gamma
  • MART-1 Antigen
  • Melanoma
  • Mutation
  • Neoplasm Proteins
  • RNA, Messenger
  • T-Lymphocytes, Cytotoxic
  • Transfection

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed ID

  • 14609574

Additional Document Info

start page

  • 86

end page

  • 97

volume

  • 224

number

  • 2