Cancer Immunology, Immunotherapy

, Volume 62, Issue 7, pp 1149–1159 | Cite as

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

  • Elise P. Salerno
  • Sofia M. Shea
  • Walter C. Olson
  • Gina R. Petroni
  • Mark E. Smolkin
  • Chantel McSkimming
  • Kimberly A. Chianese-Bullock
  • Craig L. SlingluffJr.
Original Article

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.

Keywords

Human  T cell  Incomplete Freund’s adjuvant  Peptide vaccine  Melanoma  CIMT 2012 Integrins Melanoma vaccine 

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

Supplementary material

262_2013_1435_MOESM1_ESM.pdf (391 kb)
Supplementary material 1 (PDF 390 kb)

References

  1. 1.
    Schwartzentruber DJ, Lawson DH, Richards JM, Conry RM, Miller DM, Treisman J, Gailani F, Riley L, Conlon K, Pockaj B, Kendra KL, White RL, Gonzalez R, Kuzel TM, Curti B, Leming PD, Whitman ED, Balkissoon J, Reintgen DS, Kaufman H, Marincola FM, Merino MJ, Rosenberg SA, Choyke P, Vena D, Hwu P (2011) gp100 Peptide vaccine and interleukin-2 in patients with advanced melanoma. N Engl J Med 364(22):2119–2127. doi:10.1056/NEJMoa1012863 PubMedCrossRefGoogle Scholar
  2. 2.
    Rosenberg SA, Yang JC, Restifo NP (2004) Cancer immunotherapy: moving beyond current vaccines. Nat Med 10(9):909–915. doi:10.1038/nm1100 PubMedCrossRefGoogle Scholar
  3. 3.
    Slingluff CL Jr (2011) The present and future of peptide vaccines for cancer: single or multiple, long or short, alone or in combination? Cancer J 17(5):343–350. doi:10.1097/PPO.0b013e318233e5b2 PubMedCrossRefGoogle Scholar
  4. 4.
    Slingluff CL Jr, Petroni GR, Chianese-Bullock KA, Smolkin ME, Hibbitts S, Murphy C, Johansen N, Grosh WW, Yamshchikov GV, Neese PY, Patterson JW, Fink R, Rehm PK (2007) Immunologic and clinical outcomes of a randomized phase II trial of two multipeptide vaccines for melanoma in the adjuvant setting. Clin Cancer Res 13(21):6386–6395. doi:10.1158/1078-0432.CCR-07-0486 PubMedCrossRefGoogle Scholar
  5. 5.
    Freund J, Thomson KJ (1945) A simple, rapid technic of preparing water-in-oil emulsions of penicillin, drugs and biologics. Science 101(2627):468–469. doi:10.1126/science.101.2627.468-a PubMedCrossRefGoogle Scholar
  6. 6.
    Schaefer JT, Patterson JW, Deacon DH, Smolkin ME, Petroni GR, Jackson EM, Slingluff CL Jr (2010) Dynamic changes in cellular infiltrates with repeated cutaneous vaccination: a histologic and immunophenotypic analysis. J Transl Med 8:79. doi:10.1186/1479-5876-8-79 PubMedCrossRefGoogle Scholar
  7. 7.
    Lang A, Nikolich-Zugich J (2005) Development and migration of protective CD8+ T cells into the nervous system following ocular herpes simplex virus-1 infection. J Immunol 174(5):2919–2925PubMedGoogle Scholar
  8. 8.
    Shomer NH, Fox JG, Juedes AE, Ruddle NH (2003) Helicobacter-induced chronic active lymphoid aggregates have characteristics of tertiary lymphoid tissue. Infect Immun 71(6):3572–3577. doi:10.1128/Iai.71.6.3572-3577.2003 PubMedCrossRefGoogle Scholar
  9. 9.
    Weninger W, Carlsen HS, Goodarzi M, Moazed F, Crowley MA, Baekkevold ES, Cavanagh LL, von Andrian UH (2003) Naive T cell recruitment to nonlymphoid tissues: a role for endothelium-expressed CC chemokine ligand 21 in autoimmune disease and lymphoid neogenesis. J Immunol 170(9):4638–4648PubMedGoogle Scholar
  10. 10.
    Harris RC, Chianese-Bullock KA, Petroni GR, Schaefer JT, Brill LB 2nd, Molhoek KR, Deacon DH, Patterson JW, Slingluff CL Jr (2012) The vaccine-site microenvironment induced by injection of incomplete Freund’s adjuvant, with or without melanoma peptides. J Immunother 35(1):78–88. doi:10.1097/CJI.0b013e31823731a4 PubMedCrossRefGoogle Scholar
  11. 11.
    Hailemichael Y, Dai Z, Jaffarzad N, Ye Y, Medina MA, Huang XF, Dorta-Estremera SM, Greeley NR, Nitti G, Peng W, Liu C, Lou Y, Wang Z, Ma W, Rabinovich B, Schluns KS, Davis RE, Hwu P, Overwijk WW (2013) Persistent antigen at vaccination sites induces tumor-specific CD8+ T cell sequestration, dysfunction and deletion. Nat Med 19(4):465–472. doi:10.1038/nm.3105 PubMedCrossRefGoogle Scholar
  12. 12.
    Slingluff CL Jr, Petroni GR, Chianese-Bullock KA, Smolkin ME, Ross MI, Haas NB, von Mehren M, Grosh WW (2011) Randomized multicenter trial of the effects of melanoma-associated helper peptides and cyclophosphamide on the immunogenicity of a multipeptide melanoma vaccine. J Clin Oncol 29(21):2924–2932. doi:10.1200/JCO.2010.33.8053 PubMedCrossRefGoogle Scholar
  13. 13.
    Slingluff CL Jr, Petroni GR, Olson WC, Smolkin ME, Ross MI, Haas NB, Grosh WW, Boisvert ME, Kirkwood JM, Chianese-Bullock KA (2009) Effect of granulocyte/macrophage colony-stimulating factor on circulating CD8+ and CD4+ T-cell responses to a multipeptide melanoma vaccine: outcome of a multicenter randomized trial. Clin Cancer Res 15(22):7036–7044. doi:10.1158/1078-0432.CCR-09-1544 PubMedCrossRefGoogle Scholar
  14. 14.
    Sallusto F, Geginat J, Lanzavecchia A (2004) Central memory and effector memory T cell subsets: function, generation, and maintenance. Annu Rev Immunol 22:745–763. doi:10.1146/annurev.immunol.22.012703.104702 PubMedCrossRefGoogle Scholar
  15. 15.
    Brinckerhoff LH, Kalashnikov VV, Thompson LW, Yamshchikov GV, Pierce RA, Galavotti HS, Engelhard VH, Slingluff CL Jr (1999) Terminal modifications inhibit proteolytic degradation of an immunogenic MART-1(27–35) peptide: implications for peptide vaccines. Int J Cancer 83(3):326–334PubMedCrossRefGoogle Scholar
  16. 16.
    Bijker MS, van den Eeden SJ, Franken KL, Melief CJ, Offringa R, van der Burg SH (2007) CD8+ CTL priming by exact peptide epitopes in incomplete Freund’s adjuvant induces a vanishing CTL response, whereas long peptides induce sustained CTL reactivity. J Immunol 179(8):5033–5040PubMedGoogle Scholar
  17. 17.
    Le Floc’h A, Jalil A, Franciszkiewicz K, Validire P, Vergnon I, Mami-Chouaib F (2011) Minimal engagement of CD103 on cytotoxic T lymphocytes with an E-cadherin-Fc molecule triggers lytic granule polarization via a phospholipase Cgamma-dependent pathway. Cancer Res 71(2):328–338. doi:10.1158/0008-5472.CAN-10-2457 PubMedCrossRefGoogle Scholar
  18. 18.
    Chapman TJ, Topham DJ (2010) Identification of a unique population of tissue-memory CD4+ T cells in the airways after influenza infection that is dependent on the integrin VLA-1. J Immunol 184(7):3841–3849. doi:10.4049/jimmunol.0902281 PubMedCrossRefGoogle Scholar
  19. 19.
    Franciszkiewicz K, Le Floc’h A, Jalil A, Vigant F, Robert T, Vergnon I, Mackiewicz A, Benihoud K, Validire P, Chouaib S, Combadiere C, Mami-Chouaib F (2009) Intratumoral induction of CD103 triggers tumor-specific CTL function and CCR5-dependent T-cell retention. Cancer Res 69(15):6249–6255. doi:10.1158/0008-5472.CAN-08-3571 PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Elise P. Salerno
    • 1
  • Sofia M. Shea
    • 2
  • Walter C. Olson
    • 1
  • Gina R. Petroni
    • 3
  • Mark E. Smolkin
    • 3
  • Chantel McSkimming
    • 4
  • Kimberly A. Chianese-Bullock
    • 1
  • Craig L. SlingluffJr.
    • 1
    • 5
  1. 1.Division of Surgical Oncology, Department of SurgeryUniversity of VirginiaCharlottesvilleUSA
  2. 2.Department of PathologyUniversity of VirginiaCharlottesvilleUSA
  3. 3.Department of Public Health SciencesUniversity of VirginiaCharlottesvilleUSA
  4. 4.Cardiovascular Research CenterUniversity of VirginiaCharlottesvilleUSA
  5. 5.UVA Health SystemCharlottesvilleUSA

Personalised recommendations