In vitro generation of influenza-specific polyfunctional CD4+ T cells suitable for adoptive immunotherapy

Cytotherapy. 2012 Feb;14(2):182-93. doi: 10.3109/14653249.2011.613932. Epub 2011 Sep 28.

Abstract

Background aims: Influenza viruses cause potentially fatal respiratory infections in stem cell transplant patients. Specific T cells provide long-lived host adaptive immunity to influenza viruses, and the potential for generating such cells for clinical use was investigated.

Methods: The inactivated influenza vaccine (Fluvax) approved for human use was used as the antigen source. Monocyte-derived dendritic cells pulsed with Fluvax were used to stimulate autologous peripheral blood mononuclear cells (PBMC) on days 0 and 7. Cells were expanded with interleukin (IL)-2 from day 7 onwards. Cell numbers and phenotype were assessed on day 21. The presence of influenza virus-specific cells was assessed by cytokine production and proliferative responses following restimulation with influenza antigens.

Results: Over 21 days of culture, a mean fold increase of 26.3 in cell number was observed (n = 7). Cultures were predominantly effector and central memory CD4+ cells, and expressed a phenotype characteristic of activated antigen-specific cells capable of B-cell helper function. Cytotoxic CD4+ and CD8+ cells specific for influenza and a high percentage of CD4+ cells specific for each of three influenza viruses targeted by Fluvax (H1N1, H3N2 and Brisbane viruses) were generated. In addition, T cells expanded when restimulated with antigens derived from influenza viruses.

Conclusions: We have demonstrated a clinically usable method for producing influenza virus-specific T cells that yield high numbers of highly reactive CD4+ cells suitable for adoptive immunotherapy. We propose that reconstructing host immunity through adoptive transfer of influenza virus-specific T cells will reduce the frequency of influenza-related deaths in the period of severe immune suppression that follows stem cell transplantation.

MeSH terms

  • CD4-Positive T-Lymphocytes / cytology*
  • CD4-Positive T-Lymphocytes / immunology*
  • Cell Proliferation
  • Cytokines / immunology
  • Dendritic Cells / immunology
  • Humans
  • Immunotherapy, Adoptive*
  • Influenza Vaccines / therapeutic use
  • Influenza, Human / immunology*
  • Influenza, Human / therapy
  • Interleukin-2 / immunology
  • Interleukin-2 / metabolism
  • Monocytes / immunology
  • Orthomyxoviridae / immunology
  • Vaccines, Inactivated / therapeutic use

Substances

  • Cytokines
  • Influenza Vaccines
  • Interleukin-2
  • Vaccines, Inactivated