Cancer Immunology, Immunotherapy

, Volume 57, Issue 9, pp 1381–1390 | Cite as

CD54 is a surrogate marker of antigen presenting cell activation

  • N. A. Sheikh
  • L. A. Jones
Original Article


There is no single universally accepted hallmark of antigen presenting cell (APC) activation. Instead a variety of methods are used to identify APCs and assess their activation state. These activation measures include phenotypic methods [e.g., assessing the increased expression of surface markers such as major histocompatability (MHC) class II] and functional assays (e.g., evaluating the enhanced ability to take up and process antigen, or stimulate naïve T cells). Sipuleucel-T is an investigational autologous active cellular immunotherapy product designed to stimulate a T cell immune response against human prostatic acid phosphatase (PAP), an antigen highly expressed in prostate tissue. Sipuleucel-T consists of peripheral blood mononuclear cells (PBMCs), including activated APCs displaying epitopes of PAP. In order to develop a robust reproducible potency assay that is not hampered by MHC restriction we have developed a method to simply assess the biological activation of antigen presenting cells (APCs). In the course of sipuleucel-T characterization, we analyzed various phenotypic and functional parameters to define the activation state of APCs obtained from peripheral blood. Flow cytometric assays revealed that CD54+ cells are responsible for antigen uptake, and that expression of CD54 predominantly localizes to APCs. Costimulation, as measured by an allogeneic mixed lymphocytic reaction (alloMLR) assay, showed that activity was restricted to the CD54+ cell population. Similarly, CD54+ cells harbor all of the PAP-specific antigen presentation activity, as assayed using a PAP-specific HLA-DRβ1-restricted T cell hybridoma. Finally we show that CD54 expression is substantially and consistently upregulated on APCs during culture with a GM-CSF fusion protein, and that this upregulation activity can be quantified. Thus these data support the use of CD54 upregulation as a surrogate for assessing human APC activation and validates its utility as a potency measure of sipuleucel-T.


Activation Antigen processing APC CD54 Upregulation 



The authors acknowledge the technical assistance and critical evaluation provided by Dirk Windgassen, Kajsa Jungaro and Mickey Emde.


  1. 1.
    Adams DO, Hamilton TA (1984) The cell biology of macrophage activation. Annu Rev Immunol 2:283–318PubMedCrossRefGoogle Scholar
  2. 2.
    Burch PA, Breen JK, Buckner JC, Gastineau DA, Kaur JA, Laus RL, Padley DJ, Peshwa MV, Pitot HC, Richardson RL, Smits BJ, Sopapan P, Strang G, Valone FH, Vuk-Pavlovic S (2000) Priming tissue-specific cellular immunity in a phase I trial of autologous dendritic cells for prostate cancer. Clin Cancer Res 6:2175–2182PubMedGoogle Scholar
  3. 3.
    Burch PA, Groghan GA, Gastineau DA, Jones LA, Kaur JS, Kylstra JW, Richardson RL, Valone FH, Vuk-Pavlovic S (2004) Immunotherapy (APC8015, Provenge) targeting prostatic acid phosphatase can induce durable remission of metastatic androgen-induced prostate cancer: a phase 2 trial. Prostate 60:197–204PubMedCrossRefGoogle Scholar
  4. 4.
    Carrasco YR, Fleire SJ, Cameron T, Dustin ML, Batista FD (2004) LFA-1/ICAM-1 interaction lowers the threshold of B cell activation by facilitating B cell adhesion and synapse formation. Immunity 20:58–599CrossRefGoogle Scholar
  5. 5.
    Coyle AJ, Gutierrez-Ramos JC (2001) The expanding B7 superfamily: increasing complexity in costimulatory signals regulating T cell function. Nat Immunol 2:203–209PubMedCrossRefGoogle Scholar
  6. 6.
    Dustin ML, Rothlein R, Bhan AK, Dinarello CA, Springer TA (1986) Induction by IL-1 and Interferon-γ: tissue distribution, biochemistry, and function of a natural adherence molecule (ICAM-1). J Immunol 137: 245–254PubMedGoogle Scholar
  7. 7.
    Gaglia JL, Greenfield EA, Mattoo A, Sharpe AH, Freeman GJ, Kuchroo VK (2000) Intercellular adhesion molecule 1 is critical for activation of CD28-deficient T cells. J Immunol 165:6091–6098PubMedGoogle Scholar
  8. 8.
    Gasson JC, Weisbrt RH, Kaufman SE (1984) Purified human granulocyte-macrophage colony-simulating factor: direct action on neutrophils. Science 226:1339–1342PubMedCrossRefGoogle Scholar
  9. 9.
    Greenwald RJ, Freeman GJ, Sharpe AH (2005) The B7 family revisited. Annu Rev Immunol 23:515–548PubMedCrossRefGoogle Scholar
  10. 10.
    Hsu FJ, Benike C, Fagnoni F, Liles TM, Czerwinski D, Taidi B, Engleman EG, Levy R (1996) Vaccination of patients with B-cell lymphoma using autologous antigen-pulsed dendritic cells. Nat Med 2:52–58PubMedCrossRefGoogle Scholar
  11. 11.
    Hume D (2005) The mononuclear phagocyte system. Curr Opin Immunol 18:49–33PubMedCrossRefGoogle Scholar
  12. 12.
    Jenkinson SR, Wiliams NA, Morgan DJ (2005) The role of intracellular adhesion molecule-1/LFA-1 interactions in the generation of tumor-specific CD8+ T cell responses. J Immunol 174:3401–3407PubMedGoogle Scholar
  13. 13.
    Jones LA (2005) Part II: dendritic cancer vaccine potency tests. Biologicals 33:124–125CrossRefGoogle Scholar
  14. 14.
    Laus R, Demao YM, Ruegg CL, Shapero MH, Slagle PH, Small E, Burch P, Valone FH (2001) Dendritic cell immunotherapy of cancer: preclinical models and early clinical experience. Cancer Res Therapy Control 11:1–10Google Scholar
  15. 15.
    Lebedeva T, Dustin ML, Sykulev Y (2005) ICAM-1 co-stimulates target cells to facilitate antigen presentation. Curr Opin Immunol 17:251–258PubMedCrossRefGoogle Scholar
  16. 16.
    Loken MR, Brosman JM, Bach BA, Ault KA (1990) Establishing optimal lymphocyte gates for immunophenotyping by flow cytometry. Cytometry 11:453–459PubMedCrossRefGoogle Scholar
  17. 17.
    Parameswaran N, SureshR, Bal V, Rath S, George A (2005) Lack of ICAM-1 on APCs during T cell priming leads to poor generation of central memory cells. J Immunol 175:2201–2211PubMedGoogle Scholar
  18. 18.
    Park JW, Melisko ME, Esserrman LJ, Jones LA, Wollan JB, Sims R (2007) Treatment with autologous antigen-presenting cells activated with the human epidermal growth factor receptor (HER-2)-based antigen lapuleucel-T: results of a phase I study in immunologic and clinical activity in HER-2-overexpressing breast cancer. J Clin Oncol 25:3680–3687PubMedCrossRefGoogle Scholar
  19. 19.
    Petricciani J, Egan W, Vicari G, Furesz J, Schild G (2006) Potency assays for therapeutic live whole cell cancer vaccines. Biologicals 35:107–113PubMedCrossRefGoogle Scholar
  20. 20.
    Poudrier J, Owens T (1994) CD54/intercellular adhesion molecule 1 and major histocompatibility complex II signaling induces B cells to express interleukin 2 receptors and complements help provided through CD40 ligation. J Exp Med 179:1417–1427PubMedCrossRefGoogle Scholar
  21. 21.
    Seventeer GA, Shimizu Y, Horgan KJ, Shaw S (1990) The LFA-1 ligand ICAM-1 provides an important costimulatory signal for T cell receptor-mediated activation of resting T cells. J Immunol 144:4579–4586Google Scholar
  22. 22.
    Shankar G, Bader R, Lodge PA (2004) The COSTIM bioassay: a novel potency test for dendritic cells. J Immunol Methods 285:293–299PubMedCrossRefGoogle Scholar
  23. 23.
    Shi Y, Liu CH, Roberts AI, Das J, Xu G, Ren G, Zhang Y,Zhang L, Yuan ZR, Sheng H, Tan W, Das G, Devadas S (2006) Granulocyte-macrophage colony-stimulating factor (GM-CSF) and T-cell responses: what we do and don’t know. Cell Res 16:126–133PubMedCrossRefGoogle Scholar
  24. 24.
    Small EJ, Fratesi P, Reese DM, Strang G, Laus R, Peshwa MV, Valone VH (2000) Immunotherapy of hormone-refractory prostate cancer with antigen-loaded dendritic cells. J Clin Oncol 18:3894–3903PubMedGoogle Scholar
  25. 25.
    Small EJ, Schelhammer PF, Higano CS, Redfern CH, Neumanaitis JJ, Valone FH, Verjee SS, Hershberg RM (2006) Placebo-controlled phase III trial of immunologic therapy with sipuleucel-T (APC8015) in patients with metastatic, asymptomatic hormone refractory prostate cancer. J Clin Oncol 24:3089–3094PubMedCrossRefGoogle Scholar
  26. 26.
    Somersalo K, Anikeeva N, Sims TN, Thomas VK, Strong RK, Spies T, Sykulev Y, Dustin ML (2004) Cytotoxic T lypmhocytes form an antigen-independent ring junction. J Clin Invest 113:49–57PubMedGoogle Scholar
  27. 27.
    Steinman RM, Witmer MD (1978) Lymphoid dendritic cells are potent stimulators of the primary mixed leukocyte reaction in mice. Proc Natl Acad Sci USA 75:5132–5136PubMedCrossRefGoogle Scholar
  28. 28.
    Takeda K, Akira S (2004) TLR signaling pathways. Semin Immunol 16:3–9PubMedCrossRefGoogle Scholar
  29. 29.
    Tohma S, Ramberg JE, Lipsky PE (1992) Expression and distribution of CD11a/CD18 and CD54 during human T cell-B cell interactions. J Leukoc Biol 52:97–103PubMedGoogle Scholar
  30. 30.
    Tosi D, Valenti R, Cova A, Sovena G, Huber V, Pilla L, Arienti F, Belardelli F, Parmiani G, Rivoltini L (2004) Role of cross-talk between IFNα-induced monocyte derived dendritic cells and NK cells in priming CD8+ T cell responses against human tumor antigens. J Immunol 172:5363–5370PubMedGoogle Scholar
  31. 31.
    Vidovic D, Graddis TJ, Stepan LP, Zaller DM, Laus R (2003) Specific stimulation of MHC-transgenic mouse T-cell hybridomas with xenogeneic APC. Human Immunol 64:238–244CrossRefGoogle Scholar
  32. 32.
    Wing EJ, Magee DM, Whiteside TL, Kaplan SS, Shadduck RK (1989) Recombinant human granulocyte/macrophage colony stimulating factor enhances monocyte cytotoxicity and secretion of tumor necrosis factor α and interferon in cancer patients. Blood 73:643–646PubMedGoogle Scholar

Online references

  1. 1.
    From a presentation at the Cellular, Tissue and Gene Therapies Advisory Committee meeting. January 2007 posting date. Gavin D. Perspectives on potency assays for complex biological products. Food and Drug Administration, Center for Biologics Research and Evaluation, Office of Cellular, Tissue and gene Therapy.

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Dendreon CorporationSeattleUSA

Personalised recommendations