Activation, dysfunction and retention of T cells in vaccine sites after injection of incomplete Freund's adjuvant, with or without peptide

Cancer Immunol Immunother. 2013 Jul;62(7):1149-59. doi: 10.1007/s00262-013-1435-5. Epub 2013 May 9.


We conducted a randomized clinical trial in 45 patients with resected AJCC stage IIB-IV melanoma to characterize cellular and molecular events at sites of immunization with incomplete Freund's adjuvant (IFA) alone, or a melanoma vaccine in IFA. At a primary vaccine site, all patients received a multi-peptide melanoma vaccine in IFA. At a replicate vaccine site, which was biopsied, group 1 received IFA only; group 2 received vaccine in IFA. Lymphocytes isolated from replicate vaccine site microenvironments (VSME) were compared to time-matched peripheral blood mononuclear cells (PBMC) in ELISpot and flow cytometry assays. Compared to PBMC, the VSME had fewer naïve and greater proportions of effector memory CD8(+) T cells (TCD8). The vast majority of TCD8 within the VSME were activated (CD69(+)), with a concentration of antigen-specific (tetramer(pos)) cells in the VSME, particularly in vaccine sites with peptide (group 2). CXCR3(+) lymphocytes were concentrated in the VSME of all patients, suggesting IFA-induced chemokine recruitment. TCD8 expression of retention integrins αEβ7 and α1β1 was elevated in VSME, with the highest levels observed in antigen-specific cells in VSME containing peptide (group 2). TCD8 retained in the VSME of both groups were strikingly dysfunctional, with minimal IFN-γ production in response to peptide stimulation and few tetramer(pos) cells producing IFN-γ. These data suggest that vaccine-induced selective retention and dysfunction of antigen-specific TCD8 within VSME may represent a significant mechanism underlying transient immune responses and low clinical response rates to peptide vaccines administered in IFA.

Publication types

  • Clinical Trial
  • Randomized Controlled Trial
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / metabolism
  • Antigens, Differentiation, T-Lymphocyte / metabolism
  • CD8-Positive T-Lymphocytes / immunology*
  • Cancer Vaccines / administration & dosage
  • Cancer Vaccines / immunology*
  • Female
  • Freund's Adjuvant / administration & dosage
  • Freund's Adjuvant / immunology*
  • Humans
  • Integrin alpha1beta1 / metabolism
  • Integrins / metabolism
  • Interferon-gamma / metabolism
  • Lectins, C-Type / metabolism
  • Leukocytes, Mononuclear / immunology*
  • Lipids / administration & dosage
  • Lipids / immunology*
  • Lymphocyte Activation
  • Male
  • Melanoma / immunology*
  • Melanoma / metabolism
  • Middle Aged
  • Peptides / immunology*
  • Receptors, CXCR3 / metabolism


  • Antigens, CD
  • Antigens, Differentiation, T-Lymphocyte
  • CD69 antigen
  • CXCR3 protein, human
  • Cancer Vaccines
  • Integrin alpha1beta1
  • Integrins
  • Lectins, C-Type
  • Lipids
  • Peptides
  • Receptors, CXCR3
  • incomplete Freund's adjuvant
  • integrin alphaEbeta7
  • Interferon-gamma
  • Freund's Adjuvant