Immunization with wild-type p53 gene sequences coadministered with Flt3 ligand induces an antigen-specific type 1 T-cell response

P. Parajuli, V. Pisarev, J. Sublet, A. Steffel, M. Varney, Rakesh K Singh, D. Laface, James E Talmadge

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Abstract

We examined the ability of immunization with sequential adenovirus/plasmid DNA vectors expressing human wild-type p53 to stimulate a type 1 T-cell response and induce protection against challenge from a metastatic tumor that expresses mutated murine p53. We found that tumor protection and an antigen (Ag)-specific immune response were enhanced by prior injection of Flt3 ligand (Flt3L) at a dose and schedule that significantly increased dendritic cell (DC) number and frequency. Preliminary studies using enzyme-linked immunospot and Winn assays suggested that Ag-specific CD8 cells, with their significant increase in IFN-γ-secreting activity (Tc1 cells), were responsible for the tumor protection. The delayed-type hypersensitivity response to p53 was increased in mice immunized with p53 alone or p53 and Flt3L compared with a negative control. In contrast, spleen cells from mice immunized with p53 and Flt3L exhibited a higher Ag-specific proliferative response than mice immunized with p53 alone. The frequencies of Ag-specific IFN-γ and interleukin (IL)-4-secreting cells were determined using an enzyme-linked immunospot assay, which demonstrated that the frequency of IFN-γ-secreting cells was significantly higher in mice immunized with p53 and Flt3L than in mice receiving Flt3L, excipient, or p53 treatment alone. In contrast, the frequency of IL-4-secreting cells did not differ significantly among these groups. We also observed an increased frequency of IL-12 and IFN-γ-secreting cells (but not IL-4 or IL-10) in the spleens of mice immediately after 10 days of Flt3L treatment, which was also the day of p53 priming. This observation supports the likelihood that there are multiple mechanisms of Flt3L adjuvant activity, including expansion of DC and type 1 T-cell number. Overall, these results suggest that immunization with p53 genetic sequences after in vivo expansion of DC, using Flt3L, provides a useful strategy to induce p53-specific, and protective, type 1 T-cell responses.

Original languageEnglish (US)
Pages (from-to)8227-8234
Number of pages8
JournalCancer Research
Volume61
Issue number22
StatePublished - Nov 15 2001

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p53 Genes
Immunization
T-Lymphocytes
Antigens
Interleukin-4
Dendritic Cells
Enzyme-Linked Immunospot Assay
Spleen
Cell Count
CD8 Antigens
flt3 ligand protein
Excipients
Delayed Hypersensitivity
Neoplasm Antigens
Interleukin-12
Adenoviridae
Interleukin-10
Neoplasms
Appointments and Schedules
Plasmids

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Immunization with wild-type p53 gene sequences coadministered with Flt3 ligand induces an antigen-specific type 1 T-cell response. / Parajuli, P.; Pisarev, V.; Sublet, J.; Steffel, A.; Varney, M.; Singh, Rakesh K; Laface, D.; Talmadge, James E.

In: Cancer Research, Vol. 61, No. 22, 15.11.2001, p. 8227-8234.

Research output: Contribution to journalArticle

Parajuli, P. ; Pisarev, V. ; Sublet, J. ; Steffel, A. ; Varney, M. ; Singh, Rakesh K ; Laface, D. ; Talmadge, James E. / Immunization with wild-type p53 gene sequences coadministered with Flt3 ligand induces an antigen-specific type 1 T-cell response. In: Cancer Research. 2001 ; Vol. 61, No. 22. pp. 8227-8234.
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AU - Steffel, A.

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AU - Singh, Rakesh K

AU - Laface, D.

AU - Talmadge, James E

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