Amplification of low-frequency antiviral CD8 T cell responses using autologous dendritic cells

Marie Larsson, David T. Wilkens, Jean François Fonteneau, Thomas J. Beadle, Melissa J. Merritt, Rhonda G. Kost, Patrick A.J. Haslett, Susan Cu-Uvin, Nina Bhardwaj, Douglas F. Nixon, Barbara L. Shacklett

Research output: Contribution to journalArticlepeer-review

40 Scopus citations


Objective: To utilize the potent antigen-presenting capacity of mature dendritic cells (MDC) in order to develop a rapid, sensitive method for quantifying antigen-specific CD8 T cells present at low frequency in peripheral blood. Design: Peripheral blood mononuclear cells (PBMC) were obtained from seven HIV-1-positive individuals with low to moderate CD8 T cell responses, including five on highly active antiretroviral therapy (HAART). IFN-γ ELISPOT assays were performed using either monocytes or MDC to present antigens expressed by recombinant vaccinia viruses (r-VV). Methods: Peripheral blood-derived monocytes were cultured for 5-6 days in the presence of IL-4 and granulocyte macrophage colony-stimulating factor, then matured in monocyte-conditioned medium. MDC were infected with r-VV and co-cultured in an ELISPOT assay with autologous monocyte-depleted PBMC. Results: Relative to autologous monocytes, MDC amplified detection of antigen-specific CD8 T cells by 2-30-fold in response to antigens from HIV-1, Epstein-Barr virus and cytomegalovirus. Furthermore, antigenic specificities were revealed that had not been detected using standard ELISPOT of PBMC. Conclusion: This assay will prove useful for the detection of memory T cells present at low frequency, and may be of interest for identifying subdominant cytotoxic T lymphocyte epitopes. This method may have broad applications for the detection of antiviral CD8 T cell responses in patient populations in whom such responses have been difficult to detect, including HIV-1-seropositive individuals with advanced disease or undergoing HAART.

Original languageEnglish
Pages (from-to)171-180
Number of pages10
Issue number2
StatePublished - 25 Jan 2002
Externally publishedYes


  • Cytotoxic T lymphocytes
  • Dendritic cells


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