Cancer and Metastasis Reviews

, Volume 25, Issue 2, pp 233–242 | Cite as

Immune responses in the draining lymph nodes against cancer: Implications for immunotherapy

  • Suyu ShuEmail author
  • Alistair J. Cochran
  • Rong-Rong Huang
  • Donald L. Morton
  • Holden T. Maecker


Regional lymph nodes are the first site for melanoma metastases. The sentinel node (SN), on the direct lymphatic drainage pathway, which usually harbors first metastases, demonstrates significant suppression in its ability to respond to antigenic stimulation. This down-regulation of SN immunity is likely the basis of its susceptibility to tumor metastases, suggesting a potential role of the immune system in the control of malignant tumors. Despite immune dysfunction in the SN, phase II trials of systemic post-operative immunotherapy with a polyvalent melanoma vaccine developed at the John Wayne Cancer Institute showed improved 5-year overall survival in patients with melanoma metastatic to regional nodes. However, most immunotherapy clinical trials have failed to demonstrate a significant clinical response, and analyses of immune responses to tumor-associated antigens that correlate clinical responses have not been established. Therefore, refinements in assay methodologies and improvements in vaccine designs are critical to the success of cancer immunotherapy. Antigen presentation by dendritic cells (DCs) is the most potent means to initiate a T cell immunity. Dendritic cell-based immunotherapies have been vigorously attempted in the past decade. To improve the immunogenicity of cancer vaccines, we recently generated heterokaryons of DCs and tumor cells by electrofusion. The fusion hybrids retained their full antigen-presenting capacity and all natural tumor antigens. In pre-clinical animal experiments, a single injection of the DC-tumor fusion hybrids was sufficient to mediate the regression of tumors established in the lung, skin and brain. Most interestingly, successful therapy required the delivery of fusion hybrids directly into lymphoid organs such as lymph nodes. A clinical trial is now being carried out to test the immunogenicity and therapeutic effects of fusion hybrids for the treatment of metastatic melanoma.


Sentinel lymph nodes Immunosuppression Active immunotherapy Dendritic cells Electrofusion Immune monitoring 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Turk JL: Delayed Hypersensitivity. North-Holland Publishing Company, Amsterdam, John Wiley and Sons, Inc., New York, 1967, p 138Google Scholar
  2. 2.
    Hanna MG Jr, Bucana CD, Pollack VA: Immunological stimulation in situ: the acute and chronic inflammatory responses in the induction of tumor immunity. Contemp Top Immunol 10: 267–296, 1980Google Scholar
  3. 3.
    Stephenson KR, Perry-Lalley D, Griffith KD, Shu S, Chang AE: Development of antitumor reactivity in regional draining lymph nodes from tumor-immunized and tumor-bearing murine hosts. Surgery 105: 523–528, 1989PubMedGoogle Scholar
  4. 4.
    Cochran AJ, Morton DL, Stern S, Lana AM, Essner R, Wen DR: Sentinel lymph nodes show profound downregulation of antigen-presenting cells of the paracortex: Implications for tumor biology and treatment. Mod Pathol 14: 604–608, 2001PubMedCrossRefGoogle Scholar
  5. 5.
    Ollila DW, Kelley MC, Gammon G, Morton DL: Overview of melanoma vaccines: Active specific immunotherapy for melanoma patients. Seminars in Surg Oncol 14: 328–336, 1998CrossRefGoogle Scholar
  6. 6.
    Steinman RM, Dhodapkar M: Active immunization against cancer with dendritic cells: The near future. Int J Cancer 94: 459–473, 2001PubMedCrossRefGoogle Scholar
  7. 7.
    Ridway D: The first 1000 dendritic cell vaccines. Cancer Invest 21: 873–886, 2003CrossRefGoogle Scholar
  8. 8.
    Guo Y, Wu M, Chen H, Wang X, Liu G, Li G, Ma J, SY M-S: Effective tumor vaccine generated by fusion of hepatoma cells with activated B cells. Science 263: 518–520, 1994PubMedGoogle Scholar
  9. 9.
    Gong J, Chen D, Kashiwaba M, Li Y, Chen L, Takeuchi H, Qu H, Rowse GJ, Gendler SJ, Kufe D: Reversal of tolerance to human MUC1 antigen MUC1 transgenic mice immunized with fusions of dendritic and carcinoma cells. Proc Natl Acad Sci USA 95: 6279–6283, 1998PubMedCrossRefGoogle Scholar
  10. 10.
    Akasaki Y, Kikuchi T, Homma S, Abe T, Kofe D, Ohno T: Antitumor effect of immunizations with fusions of dendritic and glioma cells in a mouse brain tumor model. J Immunother 24: 106–113, 2001CrossRefGoogle Scholar
  11. 11.
    Gong J, Nikrui N, Chen D, Koido S, Wu Z, Tanaka Y, Cannistra S, Avigan D, Kufe D: Fusions of human ovarian carcinoma cells with autologous or allogeneic dendritic cells induce antitumor immunity. J Immunol 165: 1705–1711, 2000PubMedGoogle Scholar
  12. 12.
    Gong J, Avigan D, Chen D, Wu Z, Koido S, Kashiwaba M, Kufe D: Activation of antitumor cytotoxic T lymphocytes by fusions of human dendritic cells and breast carcinoma cells. PNAS 97: 2715–2718, 2000PubMedCrossRefGoogle Scholar
  13. 13.
    Avigan D, Vasir B, Gong J, Borges V, Wu Z, Uhl L, Atkins M, Mier J, McDermott D, Smith T, Giallambardo N, Stone C, Schadt K, Dolgoff J, Tetreault J-C, Villarroel M, Kufe D: Fusion cell vaccination of patients with metastatic breast and renal cancer induces immunological and clinical responses. Clin Cancer Res 10: 4699–4708, 2004PubMedCrossRefGoogle Scholar
  14. 14.
    Cochran AJ, Bhuta S, Paul E, Ribas A: The shifting patterns of metastatic melanoma. Clin Lab Med 4: 759–783, 2000Google Scholar
  15. 15.
    Cochran AJ, Roberts AA, Saida T: The place of lymphatic mapping and sentinel node biopsy in oncology. Int J Clin Oncol 8: 139–150, 2003PubMedCrossRefGoogle Scholar
  16. 16.
    Morton DL, Wen D-R, Wong JH, Economou JS, Cagle LA, Storm FK, Foshag LJ, Cochran AJ: Technical details of intraoperative lymphatic mapping for early stage melanoma. Arch Surg 127: 392–399, 1992PubMedGoogle Scholar
  17. 17.
    Cochran AJ, Pihl E, Wen D-R, Hoon DSB, Korn EL: Zoned immune suppression of lymph nodes draining malignant melanoma: Histologic and immunohistologic studies. J Nat Cancer Inst 78: 399–405, 1987PubMedGoogle Scholar
  18. 18.
    Hoon DSB, Korn EL, Cochran AJ: Variations in functional immunocompetence of human tumor-draining lymph nodes. Cancer Res 47: 1740–1744, 1987PubMedGoogle Scholar
  19. 19.
    Wen D-R, Hoon DSB, Cochran AJ: Variations in the immunological activity of individual lymph nodes draining human cancers assessed by spontaneous generation of migration inhibiting lymphokines. Cancer Immunol Immunother 30: 277–282, 1989PubMedCrossRefGoogle Scholar
  20. 20.
    Farzad Z, Cochran AJ, McBride WH, Grey D, Wong V, Morton DL: Phenotypic changes in lymphocytes of lymph nodes draining human cutaneous melanoma. Cancer Res 50: 3585–3588, 1990PubMedGoogle Scholar
  21. 21.
    Hoon DSB, Bowker R, Cochran AJ: Suppressor cell activity in melanoma-draining lymph nodes. Cancer Res 74: 1529–1533, 1987Google Scholar
  22. 22.
    Lana A-M, Wen D-R, Cochran AJ: The morphology, immunophenotype and distribution of paracortical dendritic leukocytes in lymph nodes regional to cutaneous melanoma. Melanoma Res 11: 1–10, 2001CrossRefGoogle Scholar
  23. 23.
    Huang RR, Wen D-R, Guo J, Giuliano A, Turner R, Nguyen M, Cochran AJ: Modulation of paracortical dendritic cells and T lymphocytes in breast cancer sentinel nodes. Breast Journal 6: 225–232, 2000PubMedCrossRefGoogle Scholar
  24. 24.
    Huang RR, Wen D-R, Itakura E, Turner R, Cochran AJ: Loss of dendritic cell complexity in sentinel nodes correlates with reduced activated T-cells. The 3rd International Sentinel Nodes Congress, Yokohama, Japan, November 16–18, 2002Google Scholar
  25. 25.
    Huang RR, Wang HJ, Lin LL, Wen D-R, Itakura E, Cochran AJ: MHC-Class II molecules expression by dendritic cells correlates with activated OPD4 + T cell in of sentinel and non-sentinel nodes from melanoma patients. Mod Pathol 17(1): 382, 2004Google Scholar
  26. 26.
    Huang RR, Paul E, Wang HJ, Lin LL, Wen D-R, Itakura E, Cochran AJ: Sentinel lymph nodes are immunosuppressed whether or not they contain metastatic melanoma. Mod Pathol 18(1): 379, 2005Google Scholar
  27. 27.
    Huang RR, Paul E, Wang HJ, Lin LL, Wen D-R, Itakura E, Cochran AJ: Vascular activation and T lymphocyte transmigration vary between sentinel and non-sentinel nodes in melanoma patients. 1st International Symposium on Cancer Metastasis and the Lymphovascular System, San Francisco, CA, April 28–30, 2005Google Scholar
  28. 28.
    Lee JH, Torisu-Itakara H, Cochran AJ, Kadison A, Huynh Y, Morton DL, Essner R: Quantitative analysis of melanoma-induced cytokine-mediated immunosuppression in melanoma sentinel nodes. Clin Cancer Res 1: 107–112, 2005Google Scholar
  29. 29.
    Leong SP, Peng M, Zhou Y-M, Vaquerano JE, Chang JWC: Cytokine profiles of sentinel lymph nodes draining the primary melanoma. Ann Surg Oncol 9: 82–87, 2002PubMedCrossRefGoogle Scholar
  30. 30.
    Vuylsteke RJCLM, Molenkamp BG, Gietema HA, van Leeuwen PAM, Wijnands PGJTB, Vos W, van Diest PJ, Scheper RJ, Meijer S, de Gruijl TD: Local administration of granulocyte/macrophage colony-stimulating factor increases the number and activation state of dendritic cells in the sentinel lymph node of early-stage melanoma. Cancer Res 64: 8456–8460, 2004PubMedCrossRefGoogle Scholar
  31. 31.
    Hirakawa S, Kodama S, Kunstfeld R, Kajiya K, Brown LF, Detmar M: VEGF-A induces tumor and sentinel lymph node lymphangiogenesis and promotes lymphatic metastasis. J Exp Med 201: 1089–1099, 2005PubMedCrossRefGoogle Scholar
  32. 32.
    Barbour AH, Coventry BJ: Dendritic cell density and activation status of tumour-infiltrating lymphocytes in metastatic human melanoma: Possible implications for sentinel node metastases. Melanoma Res 13: 263–269, 2003PubMedCrossRefGoogle Scholar
  33. 33.
    Straten PT, Dahl C, Schrama D, Pedersen L, Andersen MH, Seremet T, Brocker E-B, Guldberg P, Becker JC: Recruitment of immature plasmacytoid dendritic cells (plasmacytoid monocytes) and myeloid dendritic cells in primary cutaneous melanomas. J Pathol 200: 255–268, 2003CrossRefGoogle Scholar
  34. 34.
    Lee JH, Essner R, Torisu-Itakura H, Wanek L, Wang H-J, Morton DL: Factors predictive of tumor-positive nonsentinel lymph nodes after tumor-positive sentinel lymph node dissection for melanoma. J Clin Oncol 22: 3677–3684, 2004PubMedCrossRefGoogle Scholar
  35. 35.
    Hoon DSB, Irie RF, Cochran AJ: Gangliosides from human melanoma immunomodulate the response of T cells to interleukin-2. Cell Immunol 111: 410–419, 1988PubMedCrossRefGoogle Scholar
  36. 36.
    Hoon DSB, Jung T, Nangauyan J, Cochran AJ, Morton DL, McBride WH: Modulation of human macrophage functions by gangliosides. Immunol Letters 20: 269–276, 1989CrossRefGoogle Scholar
  37. 37.
    Itakura E, Huang RR, Wen D-R, Cochran AJ: Is sentinel node susceptibility to metastases influenced by tumor-derived cytokines? RT in situ PCR studies. Mod Pathol 17(Supple 1): 385, 2004Google Scholar
  38. 38.
    Itakura E, Huang RR, Wen D-R, Cochran AJ: “Stealth” Melanoma Cells: the basis of histology-negative PCR-positive sentinel nodes? Mod Pathol 18(1): 380, 2005Google Scholar
  39. 39.
    Berd D, MaGuire HC, McCue P, Mastrangelo MJ: Treatment of metastatic melanoma with an autologous tumor-cell vaccine: Clinical and immunologic results in 64 patients. J Clin Oncol 8: 1855–1867, 1990Google Scholar
  40. 40.
    Morton DL, Foshag LJ, Hoon DS, Nizze JA, Famatiga E, Wanek LA, Chang C, Davtyan DG, Gupta RK, Elashoff R: Prolongation of survival in metastatic melanoma after active specific immunotherapy with a new polyvalent melanoma vaccine. Ann Surg 216: 463–482, 1992PubMedGoogle Scholar
  41. 41.
    Neil G, Zimmermann U: Electrofusion. Methods Enzymol 220: 174–196, 1993PubMedCrossRefGoogle Scholar
  42. 42.
    White KL: Electrofusion of mammalian cells. In Nikoloff JA (ed) Methods in Molecular Biology. The Humana Press, Clifton, NJ, 1995, pp 283–293Google Scholar
  43. 43.
    Kuriyama H, Shimizu K, Lee W, Kjaergaard J, Parkhurst MR, Cohen PA, Shu S: Therapeutic vaccine generated by electrofusion of dendritic cells and tumour cells. Dev Biol (Basel) 116: 157–166, 2004Google Scholar
  44. 44.
    Hayashi T, Tanaka H, Tanaka J, Wang R, Averbook BJ, Cohen PA, Shu S: Immunogenicity and therapeutic efficacy of dendritic-tumor hybrid cells generated by electrofusion. Clin Immunol 104: 14–20, 2002PubMedCrossRefGoogle Scholar
  45. 45.
    Kjaergaard J, Shimizu K, Shu S: Electrofusion of syngeneic dendritic cells and tumor generates potent therapeutic vaccine. Cell Immunol 225: 65–74, 2003PubMedCrossRefGoogle Scholar
  46. 46.
    Shimizu K, Kuriyama H, Kjaergaard J, Lee W, Tanaka H, Shu S: Comparative analysis of antigen loading strategies of dendritic cells for tumor immunotherapy. J Immunother 27: 265–272, 2004PubMedCrossRefGoogle Scholar
  47. 47.
    Kjaergaard J, Wang L-X, Kuriyama H, Shu S, Plautz GE: Active immunotherapy for advanced intracranial murine tumors by using dendritic cell-tumor cell fusion vaccines. J Neurosurg 103: 156–164, 2005PubMedCrossRefGoogle Scholar
  48. 48.
    Parkhurst MR, DePan C, Riley JP, Rosenberg SA, Shu S: Hybrids of dendritic cells and tumor cells generated by electrofusion simultaneously present immunodominant epitopes from multiple human tumor-associated antigens in the context of MHC class I and class II molecules. J Immunol 170: 5317–5325, 2003PubMedGoogle Scholar
  49. 49.
    Helms T, Boehm BO, Asaad RJ, Trezza RP, Lehmann PV, Tary-Lehmann M: Diret visualization of cytokine-producing recall antigen-specific CD4 memory T cells in healthy individuals and HIV patients. J Immunol 164: 3723–3732, 2000PubMedGoogle Scholar
  50. 50.
    Suni MA, Dunn HS, Orr PL, deLaat R, Sinclair E, Chanekar SA, Bredt BM, Dunne JF, Maino VC, Maecker HT: Performance of plate-based cytokine flow cytometry with automated data analysis. BMC Immunol 4: 9, 2003PubMedCrossRefGoogle Scholar
  51. 51.
    Maecker HT, Dunn HS, Suni MA, Khatamzas E, Pitcher CJ, Bunde T, Persaud N, Trigona W, Fu TM, Sinclair E, Bredt BM, McCune JM, Maino VC, Kern F, Picker LJ: Use of overlapping peptide mixtures as antigens for cytokine flow cytometry. J Immunol Methods 255: 27–40, 2001PubMedCrossRefGoogle Scholar
  52. 52.
    Altman JD, Moss PAH, Goulder PJR, Barouch DH, McHeyzer-Williams MG, Bell JI, McMichael AJ, Davis MM: Phenotypic analysis of antigen-specific T lymphocytes. Science 274: 94–96, 1996PubMedCrossRefGoogle Scholar
  53. 53.
    Suni MA, Maino VC, Maecker HT: Ex vivo analysis of T-cell function. Curr Opin Immunol 17: 434–440, 2005PubMedCrossRefGoogle Scholar
  54. 54.
    Smith JG, Liu X, Kaufhold RM, Clair J, Caulfield MJ: Development and validation of a gamma interferon ELISPOT assay for quantitation of cellular immune responses to varicella-zoster virus. Clin Diagn Lab Immunol 8: 871–879, 2001PubMedCrossRefGoogle Scholar
  55. 55.
    Maecker HT: Cytokine flow cytometry in the analysis of tumor-specific T cells. In Kieber-Emmons T (ed) Methods in Cancer Vaccine Development. Humana Press, Totowa, NJ, in press, 2005Google Scholar
  56. 56.
    Maeker HT, Rinfret A, D'Souza P, Darden J, Roig E, Landry C, Hayes P, Birungi J, Anzala O, Garcia M, Harari A, Frank I, Baydo R, Baker M, Holbrook J, Ottinger J, Lamoreaux L, Epling CL, Sinclair E, Suni MA, Punt K, Calarota S, El-Bahi S, Alter G, Maila H, Kuta E, Cox J, Gray C, Altfeld M, Nougarede N, Boyer J, Tussey L, Tobery T, Bredt B, Roederer M, Koup R, Maino VC, Weinhold K, Pantaleo G, Gilmour J, Horton H, Sekaly RP: Standardization of cytokine flow cytometry assays. BMC Immunol 6: 13, 2005CrossRefGoogle Scholar
  57. 57.
    Campbell MJ, Scott J, Maecker HT, Park JW, Esserman LJ: Immune dysfunction and micrometastases in women with breast cancer. Breast Cancer Res Treat 91: 163–171, 2005PubMedCrossRefGoogle Scholar
  58. 58.
    Leong SP, Peng M, Zhou YM, Vaquerano JE, Chang JW: Cytokine profiles of sentinel lymph nodes draining the primary melanoma. Ann Surg Oncol 9: 82–87, 2002PubMedCrossRefGoogle Scholar
  59. 59.
    Cox JH, Ferrari G, Kalams SA, Lopaczynski W, Oden N, Group ECS: Results of an ELISPOT proficiency panel conducted in 11 laboratories participating in international human immunodeficiency virus type 1 vaccine trials. AIDS Res Hum Retroviruses 21: 68–81, 2005PubMedCrossRefGoogle Scholar
  60. 60.
    Smith JW 2nd, Walker EB, Fox BA, Haley D, Wisner KP, Doran T, Fisher B, Justice L, Wood W, Vetto J, Maecker H, Dols A, Meijer S, Hu HM, Romero P, Alvord WG, Urba WJ: Adjuvant immunization of HLA-A2-positive melanoma patients with a modified gp100 peptide induces peptide-specific CD8 + T-cell responses. J Clin Oncol 21: 1562–1573, 2003PubMedCrossRefGoogle Scholar
  61. 61.
    Maecker HT, Auffermann-Gretzinger S, Nomura LE, Liso A, Czerwinski DK, Levy R: Detection of CD4 T-cell responses to a tumor vaccine by cytokine flow cytometry. Clin Cancer Res 7: 902s–908s, 2001PubMedGoogle Scholar
  62. 62.
    Jacobson MA, Maecker HT, Orr PL, D'Amico R, Van Natta M, Li XD, Pollard RB, Bredt BM: Results of a cytomegalovirus (CMV)-specific CD8+/interferon-gamma + Cytokine flow cytometry assay correlate with clinical evidence of protective immunity in patients with AIDS with CMV retinitis. J Infect Dis 189: 1362–1372, 2004PubMedCrossRefGoogle Scholar
  63. 63.
    Pantaleo G, Koup RA: Correlates of immune protection in HIV-1 infection: What we know, what we don't know, what we should know. Nat Med 10: 806–810, 2004PubMedCrossRefGoogle Scholar
  64. 64.
    Lee KH, Wang E, Nielsen MB, Wunderlich J, Migueles S, Connors M, Steinberg SM, Rosenberg SA, Marincola FM: Increased vaccine-specific T cell frequency after peptide-based vaccination correlates with increased susceptibility to in vitro stimulation but does not lead to tumor regression. J Immunol 163: 6292–6300, 1990Google Scholar
  65. 65.
    Yee C, Savage PA, Lee PP, Davis MM, Greenberg PD: Isolation of high avidity melanoma-reactive CTL from heterogeneous populations using peptide-MHC tetramers. J Immunol 162: 2227–2234, 1999PubMedGoogle Scholar
  66. 66.
    Swetter S, Thompson J, Greenberg PD, Roederer M, Davis MM: Characterization of circulating T cells specific for tumor-associated antigens in melanoma patients. Nat Med 5: 677–685, 1999PubMedCrossRefGoogle Scholar
  67. 67.
    Rubio V, Stuge TB, Singh N, Betts MR, Weber JS, Roederer M, Lee PP: Ex vivo identification, isolation and analysis of tumor-cytolytic T cells. Nat Med 9: 1377–1382, 2003PubMedCrossRefGoogle Scholar
  68. 68.
    Walker EB, Haley D, Miller W, Floyd K, Wisner KP, Sanjuan N, Maecker HT, Romero P, Hu HM, Alvord WG, Smith JW, Fox BA, Urba WJ: gp100 (209-2M) peptide immunization of HLA-A2+ stage I–III melanoma patients induces significant increase in antigen-specific effector and long-term memory CD8 + T cells. Clin Cancer Res 10: 668–680, 2004PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science + Business Media, LLC 2006

Authors and Affiliations

  • Suyu Shu
    • 1
    Email author
  • Alistair J. Cochran
    • 2
  • Rong-Rong Huang
    • 2
  • Donald L. Morton
    • 3
  • Holden T. Maecker
    • 4
  1. 1.Center for Surgery ResearchThe Cleveland Clinic FoundationClevelandUSA
  2. 2.Departments of Pathology and Laboratory and SurgeryDavid Geffen School of Medicine, UCLALos AngelesUSA
  3. 3.John Wayne Cancer InstituteSanta MonicaUSA
  4. 4.BD BioscienceSan JoseUSA

Personalised recommendations