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

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Abstract

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 (tetramerpos) 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 tetramerpos 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.

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Abbreviations

CLA:

Cutaneous leukocyte antigen

HEV:

High endothelial venule

IFA:

Incomplete Freund’s adjuvant

PBMC:

Peripheral blood mononuclear cell

TCM :

Central memory T cell (CD45RAneg/CCR7+)

TEM :

Effector memory T cell (CD45RAneg/CCR7neg)

TEMRA :

CD45RApos effector memory T cell (CD45RAneg/CCR7neg)

TLO:

Tertiary lymphoid organ

Tnaive :

Naïve T cell (CD45RA+/CCR7+)

VSME:

Vaccine site microenvironment

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Acknowledgments

We greatly appreciate the work of Patrice Neese, Carmel Nail, and Priscilla Merrill in care of the patients on this trial, and Cheryl F.Murphy, Nadejda V. Galeassi, Kelly Smith and Elizabeth Coleman in immune analyses and data management. Funding support was provided by the National Institutes of Health Research Training Grant T32HL007849 (PI- Irving Kron) to EPS and the University of Virginia Rebecca Clay Harris Memorial Fellowship to SMS. Additional funding for this study was provided by the University of Virginia Cancer Center Support Grant (NIH/NCI P30 CA44579, Biorepository and Tissue Research Facility) and the University of Virginia General Clinical Research Center (NIH M01 RR00847). Peptides used in this vaccine were prepared with philanthropic support from the Commonwealth Foundation for Cancer Research and Alice and Bill Goodwin. Additional philanthropic support was provided by the James and Rebecca Craig Foundation, George S. Suddock, Richard and Sherry Sharp, and the Patients and Friends Research Funds of the University of Virginia Cancer Center. Montanide ISA-51 (produced by Seppic, Inc.) was used in the vaccines of this trial, but paid for by the University of Virginia. No corporate funding support was provided for this study.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Division of Surgical Oncology, Department of Surgery, University of Virginia, Charlottesville, VA, USA

    Elise P. Salerno, Walter C. Olson, Kimberly A. Chianese-Bullock & Craig L. Slingluff Jr.

  2. Department of Pathology, University of Virginia, Charlottesville, VA, USA

    Sofia M. Shea

  3. Department of Public Health Sciences, University of Virginia, Charlottesville, VA, USA

    Gina R. Petroni & Mark E. Smolkin

  4. Cardiovascular Research Center, University of Virginia, Charlottesville, VA, USA

    Chantel McSkimming

  5. UVA Health System, Carter Harrison Research Building, G713, P.O. Box 800709, Charlottesville, VA, 22908-0709, USA

    Craig L. Slingluff Jr.

Authors
  1. Elise P. Salerno
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  2. Sofia M. Shea
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  3. Walter C. Olson
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  4. Gina R. Petroni
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  5. Mark E. Smolkin
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  6. Chantel McSkimming
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  7. Kimberly A. Chianese-Bullock
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  8. Craig L. Slingluff Jr.
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Corresponding author

Correspondence to Craig L. Slingluff Jr..

Additional information

Elise P. Salerno and Sofia M. Shea contributed equally to this work.

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Salerno, E.P., Shea, S.M., Olson, W.C. et al. Activation, dysfunction and retention of T cells in vaccine sites after injection of incomplete Freund’s adjuvant, with or without peptide. Cancer Immunol Immunother 62, 1149–1159 (2013). https://doi.org/10.1007/s00262-013-1435-5

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  • DOI: https://doi.org/10.1007/s00262-013-1435-5

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