Use of recombinant poxviruses to stimulate anti-melanoma T cell reactivity

Christina J. Kim, Janice Cormier, Matthew Roden, Linda Gritz, Gail P. Mazzara, Patricia Fetsch, Yasmine Hijazi, Kang Hun Lee, Steven A. Rosenberg, Francesco M. Marincola

Research output: Contribution to journalArticle

Abstract

Background: Dendritic cells (DC) are potent professional antigen- presenting cells that can activate naive T lymphocytes and initiate cellular immune responses. As adjuvants, DC may be useful for enhancing immunogenicity and mediating tumor regression. Endogenous expression of antigen by DC could offer the potential advantage of allowing prolonged constitutive presentation of endogenously processed epitopes and exploitation of multiple restriction elements for the presentation of the same antigen. Methods: DC were prepared from the peripheral blood of HLA A*0201 patients with metastatic melanoma in the presence of IL-4 (1000 IU/mL) and GMCSF (1000 IU/mL). Recombinant vaccinia and fowlpox viruses encoding the hMART-1 gene were constructed and used to infect DC. The efficiency of infection and expression of the MART-1 antigen were assessed by immunohistochemistry and intracellular FACS analyses. Cytotoxic lymphocytes (CTL) were generated by the stimulation of CD8+ T cells, with DC expressing the recombinant gene. Reactivity of the CTL was determined at weeks 1 and 2 by the amount of IFN-γ released. Results: DC were infected with recombinant poxviruses and demonstrated specific melanoma antigen expression by immunohistochemistry, immunofluorescence, and intracellular FACS analysis. The expression by DC of MART-1 MAA after vital infection was sufficient to generate CD8+ T lymphocytes that recognized naturally processed epitopes on tumor cells in 10 of 11 patients. Conclusions: Human DC are receptive to infection by recombinant poxviruses encoding MAA genes and are capable of efficiently processing and presenting these MAA to cytotoxic T cells. The potential advantage of this approach is the ability to present specific antigen independent of the identification of the epitope or the MHC restriction element. This strategy may be useful for the identification of relevant epitopes for a diverse number of HLA alleles and for active immunization in patients.

Original languageEnglish (US)
Pages (from-to)64-76
Number of pages13
JournalAnnals of Surgical Oncology
Volume5
Issue number1
DOIs
StatePublished - Jan 1 1998

Keywords

  • CTL
  • Dendritic cells
  • MART- 1
  • Melanoma
  • Melanoma-associated antigens
  • Poxviruses
  • Recombinant viral vectors

ASJC Scopus subject areas

  • Surgery
  • Oncology

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