Cellular immunity of patients with malignant glioma: prerequisites for dendritic cell vaccination immunotherapy

View More View Less
  • 1 Department of Neurosurgery and Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University Medical Center, Düsseldorf, Germany
Restricted access

Purchase Now

USD  $45.00

JNS + Pediatrics - 1 year subscription bundle (Individuals Only)

USD  $505.00

JNS + Pediatrics + Spine - 1 year subscription bundle (Individuals Only)

USD  $600.00
Print or Print + Online

Object

Vaccination therapy that uses dendritic cells (DCs) is a promising immunotherapeutic approach. However, it relies on intact cellular immunity and efficient generation of mature DCs, both of which can be impaired in patients with glioma. Therefore, the immune status and ex vivo generation of DC in such patients were studied.

Methods

The frequencies of white blood cell subsets and monocyte-derived, mature DCs in patients with high-grade gliomas and healthy control volunteers were analyzed using flow cytometry.

In the patients, frequencies of lymphocytes, T cells, and B cells were reduced in comparison with the volunteers in the control group, whereas frequencies of neutrophils and monocytes were increased. There were no differences between the two groups in terms of white blood cell counts or the frequency of NK cells and the major T-cell subsets. The responsiveness of T cells to lectin stimulation was normal. For monocytes, lower frequencies of CD80+ and CD86+ cells but not of CD40+ and HLA-DR+ cells were observed in patients. Ex vivo DC generation in a two-step culture protocol in autologous plasma–supplemented medium or in serum-free medium showed only minor differences in CD80 and HLA-DR expression between the patient and control groups. Frequencies of CD83+, CD1a+, CD14, CD40+, and CD86+ cells were comparable. Overall, the serum-free medium was superior to the plasma-supplemented medium and allowed efficient ex vivo generation of CD83+, CD1a+, and CD14 mature DCs.

Conclusions

Only minor defects in the immune status of patients with glioma were observed, which probably would not hamper immunotherapy. Mature DCs can be generated successfully in normal numbers and with typical immunophenotypes from monocytes of patients with glioma, particularly under serum-free conditions.

Abbreviations used in this paper:BrdU = bromodeoxyuridine; DC = dendritic cell; FITC = fluorescein isothiocyanate; GM-CSF = granulocyte–macrophage colony-stimulating factor; GMP = good manufacturing practice; HLA-DR = human leukocyte antigen, locus DR; IL = interleukin; mAb = monoclonal antibody; NK = natural killer; PBMC = peripheral blood mononuclear cell; PHA = phytohemagglutinin; PHA-M = the mucoprotein form of PHA; SEM = standard error of the mean; TNFα= tumor necrosis factor–α.

JNS + Pediatrics - 1 year subscription bundle (Individuals Only)

USD  $505.00

JNS + Pediatrics + Spine - 1 year subscription bundle (Individuals Only)

USD  $600.00

Contributor Notes

Address reprint requests to: Rüdiger V. Sorg, Ph.D., Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University Medical Center, Moorenstrasse 5, Bldg. 14.80, 40225 Düsseldorf, Germany. email: rsorg@itz.uni-duesseldorf.de.
  • 1

    Ashkenazi E, , Deutsch M, , Tirosh R, , Weinreb A, , Tsukerman A, & Brodie C: A selective impairment of the IL-2 system in lymphocytes of patients with glioblastomas: increased level of soluble IL-2R and reduced protein tyrosine phosphorylation. Neuroimmunomodulation 4:4956, 1997

    • Search Google Scholar
    • Export Citation
  • 2

    Ausiello C, , Maleci A, , Spagnoli GC, , Antonelli G, & Cassone A: Cell-mediated cytotoxicity in glioma-bearing patients: differential responses of peripheral blood mononuclear cells to stimulation with interleukin-2 and microbial antigen. J Neurooncol 6:329338, 1988

    • Search Google Scholar
    • Export Citation
  • 3

    Banchereau J, , Briere F, , Caux C, , Davoust J, , Lebecque S, & Liu YJ, : Immunobiology of dendritic cells. Annu Rev Immunol 18:767811, 2000

  • 4

    Bhondeley MK, , Mehra RD, , Mehra NK, , Mohapatra AK, , Tandon PN, & Roy S, : Imbalances in T cell subpopulations in human gliomas. J Neurosurg 68:589593, 1988

    • Search Google Scholar
    • Export Citation
  • 5

    Blom U, , Blomgren H, , Ullen H, , Collins P, & Von Holst H: Mitogen stimulation of blood lymphocytes from patients with primary intracranial tumors. Correlation to histological tumor type. Anticancer Res 5:343348, 1985

    • Search Google Scholar
    • Export Citation
  • 6

    Braun DP, , Penn RD, , Flannery AM, & Harris JE: Immunoregulatory cell function in peripheral blood leukocytes of patients with intracranial gliomas. Neurosurgery 10:203209, 1982

    • Search Google Scholar
    • Export Citation
  • 7

    Brooks WH, , Latta RB, , Mahaley MS, , Roszman TL, , Dudka L, & Skaggs C: Immunobiology of primary intracranial tumors. Part 5: Correlation of a lymphocyte index and clinical status. J Neurosurg 54:331337, 1981

    • Search Google Scholar
    • Export Citation
  • 8

    Burger PC, , Heinz ER, , Shibata T, & Kleihues P: Topographic anatomy and CT correlations in the untreated glioblastoma multiforme. J Neurosurg 68:698704, 1988

    • Search Google Scholar
    • Export Citation
  • 9

    Castelli MG, , Chiabrando C, , Fanelli R, , Martelli L, , Butti G, & Gaetani P, : Prostaglandin and thromboxane synthesis by human intracranial tumors. Cancer Res 49:15051508, 1989

    • Search Google Scholar
    • Export Citation
  • 10

    de Martin R, , Haendler B, , Hofer-Warbinek R, , Gaugitsch H, , Wrann M, & Schlusener H, : Complementary DNA for human glioblastoma-derived T cell suppressor factor, a novel member of the transforming growth factor-beta gene family. Embo J 6:36733677, 1987

    • Search Google Scholar
    • Export Citation
  • 11

    Dhodapkar KM, , Cirignano B, , Chamian F, , Zagzag D, , Miller DC, & Finlay JL, : Invariant natural killer T cells are preserved in patients with glioma and exhibit antitumor lytic activity following dendritic cell-mediated expansion. Int J Cancer 109:893899, 2004

    • Search Google Scholar
    • Export Citation
  • 12

    Dhodapkar MV, , Steinman RM, , Krasovsky J, , Munz C, & Bhardwaj N: Antigen-specific inhibition of effector T cell function in humans after injection of immature dendritic cells. J Exp Med 193:233238, 2001

    • Search Google Scholar
    • Export Citation
  • 13

    Dix AR, , Brooks WH, , Roszman TL, & Morford LA: Immune defects observed in patients with primary malignant brain tumors. J Neuroimmunol 100:216232, 1999

    • Search Google Scholar
    • Export Citation
  • 14

    Elliott L, , Brooks W, & Roszman T: Role of interleukin-2 (IL-2) and IL-2 receptor expression in the proliferative defect observed in mitogen-stimulated lymphocytes from patients with gliomas. J Natl Cancer Inst 78:919922, 1987

    • Search Google Scholar
    • Export Citation
  • 15

    Elliott LH, , Brooks WH, & Roszman TL: Cytokinetic basis for the impaired activation of lymphocytes from patients with primary intracranial tumors. J Immunol 132:12081215, 1984

    • Search Google Scholar
    • Export Citation
  • 16

    Elliott LH, , Brooks WH, & Roszman TL: Inability of mitogen-activated lymphocytes obtained from patients with malignant primary intracranial tumors to express high affinity interleukin 2 receptors. J Clin Invest 86:8086, 1990

    • Search Google Scholar
    • Export Citation
  • 17

    Fong L, & Engleman EG: Dendritic cells in cancer immunotherapy. Annu Rev Immunol 18:245273, 2000

  • 18

    Guinan EC, , Gribben JG, , Boussiotis VA, , Freeman GJ, & Nadler LM: Pivotal role of the B7: CD28 pathway in transplantation tolerance and tumor immunity. Blood 84:32613282, 1994

    • Search Google Scholar
    • Export Citation
  • 19

    Hintzen RQ, , Voormolen J, , Sonneveld P, & van Duinen SG: Glioblastoma causing granulocytosis by secretion of granulocyte-colony-stimulating factor. Neurology 54:259261, 2000

    • Search Google Scholar
    • Export Citation
  • 20

    Kikuchi T, , Abe T, & Ohno T: Effects of glioma cells on maturation of dendritic cells. J Neurooncol 58:125130, 2002

  • 21

    Kikuchi T, , Akasaki Y, , Irie M, , Homma S, , Abe T, & Ohno T: Results of a phase I clinical trial of vaccination of glioma patients with fusions of dendritic and glioma cells. Cancer Immunol Immunother 50:337344, 2001

    • Search Google Scholar
    • Export Citation
  • 22

    Lebecque S: Antigen receptors and dendritic cells. Vaccine 18:16031605, 2000

  • 23

    Lipscomb MF, & Masten BJ: Dendritic cells: immune regulators in health and disease. Physiol Rev 82:97130, 2002

  • 24

    Matasic R, , Dietz AB, & Vuk-Pavlovic S: Dexamethasone inhibits dendritic cell maturation by redirecting differentiation of a subset of cells. J Leukoc Biol 66:909914, 1999

    • Search Google Scholar
    • Export Citation
  • 25

    McIlroy D, & Gregoire M: Optimizing dendritic cell-based anticancer immunotherapy: maturation state does have clinical impact. Cancer Immunol Immunother 52:583591, 2003

    • Search Google Scholar
    • Export Citation
  • 26

    McVicar DW, , Davis DF, & Merchant RE: In vitro analysis of the proliferative potential of T cells from patients with brain tumor: glioma-associated immunosuppression unrelated to intrinsic cellular defect. J Neurosurg 76:251260, 1992

    • Search Google Scholar
    • Export Citation
  • 27

    Menetrier-Caux C, , Montmain G, , Dieu MC, , Bain C, , Favrot MC, & Caux C, : Inhibition of the differentiation of dendritic cells from CD34(+) progenitors by tumor cells: role of interleukin-6 and macrophage colony-stimulating factor. Blood 92:47784791, 1998

    • Search Google Scholar
    • Export Citation
  • 28

    Morford LA, , Dix AR, , Brooks WH, & Roszman TL: Apoptotic elimination of peripheral T lymphocytes in patients with primary intracranial tumors. J Neurosurg 91:935946, 1999

    • Search Google Scholar
    • Export Citation
  • 29

    Morford LA, , Elliott LH, , Carlson SL, , Brooks WH, & Roszman TL: T cell receptor-mediated signaling is defective in T cells obtained from patients with primary intracranial tumors. J Immunol 159:44154425, 1997

    • Search Google Scholar
    • Export Citation
  • 30

    Nitta T, , Hishii M, , Sato K, & Okumura K: Selective expression of interleukin-10 gene within glioblastoma multiforme. Brain Res 649:122128, 1994

    • Search Google Scholar
    • Export Citation
  • 31

    Parney IF, , Hao C, & Petruk KC: Glioma immunology and immunotherapy. Neurosurgery 46:778772, 2000

  • 32

    Piemonti L, , Monti P, , Allavena P, , Sironi M, , Soldini L, & Leone BE, : Glucocorticoids affect human dendritic cell differentiation and maturation. J Immunol 162:64736481, 1999

    • Search Google Scholar
    • Export Citation
  • 33

    Prins RM, , Graf MR, , Merchant RE, , Black KL, & Wheeler CJ: Thymic function and output of recent thymic emigrant T cells during intracranial glioma progression. J Neurooncol 64:4554, 2003

    • Search Google Scholar
    • Export Citation
  • 34

    Roth W, & Weller M: Chemotherapy and immunotherapy of malignant glioma: molecular mechanisms and clinical perspectives. Cell Mol Life Sci 56:481506, 1999

    • Search Google Scholar
    • Export Citation
  • 35

    Rutkowski S, , De Vleeschouwer S, , Kaempgen E, , Wolff JE, , Kuhl J, & Demaerel P, : Surgery and adjuvant dendritic cell-based tumor vaccination for patients with relapsed malignant glioma, a feasibility study. Br J Cancer 91:16561662, 2004

    • Search Google Scholar
    • Export Citation
  • 36

    Schuler G, , Schuler-Thurner B, & Steinman RM: The use of dendritic cells in cancer immunotherapy. Curr Opin Immunol 15:138147, 2003

  • 37

    Sorg RV, , Andres S, , Kogler G, , Fischer J, & Wernet P: Phenotypic and functional comparison of monocytes from cord blood and granulocyte colony-stimulating factor-mobilized apheresis products. Exp Hematol 29:12891294, 2001

    • Search Google Scholar
    • Export Citation
  • 38

    Sorg RV, , Özcan Z, , Brefort T, , Fischer J, , Ackermann R, & Müller M, : Clinical-scale generation of dendritic cells in a closed system. J Immunotherapy 26:374383, 2003

    • Search Google Scholar
    • Export Citation
  • 39

    Steinbrink K, , Wolfl M, , Jonuleit H, , Knop J, & Enk AH: Induction of tolerance by IL-10-treated dendritic cells. J Immunol 159:47724780, 1997

    • Search Google Scholar
    • Export Citation
  • 40

    Strobl H, & Knapp W: TGF-beta1 regulation of dendritic cells. Microbes Infect 1:12831290, 1999

  • 41

    Surawicz TS, , Davis F, , Freels S, , Laws ER Jr, & Menck HR: Brain tumor survival: results from the National Cancer Data Base. J Neurooncol 40:151160, 1998

    • Search Google Scholar
    • Export Citation
  • 42

    Van Meir E, , Sawamura Y, , Diserens AC, , Hamou MF, & de Tribolet N: Human glioblastoma cells release interleukin 6 in vivo and in vitro. Cancer Res 50:66836688, 1990

    • Search Google Scholar
    • Export Citation
  • 43

    Woiciechowsky C, , Asadullah K, , Nestler D, , Schoning B, , Glockner F, & Docke WD, : Diminished monocytic HLA-DR expression and ex vivo cytokine secretion capacity in patients with glioblastoma: effect of tumor extirpation. J Neuroimmunol 84:164171, 1998

    • Search Google Scholar
    • Export Citation
  • 44

    Wood GW, & Morantz RA: Depressed T lymphocyte function in brain tumor patients: monocytes as suppressor cells. J Neurooncol 1:8794, 1983

    • Search Google Scholar
    • Export Citation
  • 45

    Yamanaka R, , Abe T, , Yajima N, , Tsuchiya N, , Homma J, & Kobayashi T, : Vaccination of recurrent glioma patients with tumor lysate-pulsed dendritic cells elicits immune responses: results of a clinical phase I/II trial. Br J Cancer 89:11721179, 2003

    • Search Google Scholar
    • Export Citation
  • 46

    Yamanaka R, , Yajima N, , Abe T, , Tsuchiya N, , Homma J, & Narita M, : Dendritic cell-based glioma immunotherapy (review). Int J Oncol 23:515, 2003

    • Search Google Scholar
    • Export Citation
  • 47

    Yang L, , Yamagata N, , Yadav R, , Brandon S, , Courtney RL, & Morrow JD, : Cancer-associated immunodeficiency and dendritic cell abnormalities mediated by the prostaglandin EP2 receptor. J Clin Invest 111:727735, 2003

    • Search Google Scholar
    • Export Citation
  • 48

    Yang T, , Witham TF, , Villa L, , Erff M, , Attanucci J, & Watkins S, : Glioma-associated hyaluronan induces apoptosis in dendritic cells via inducible nitric oxide synthase: implications for the use of dendritic cells for therapy of gliomas. Cancer Res 62:25832591, 2002

    • Search Google Scholar
    • Export Citation
  • 49

    Yu JS, , Liu G, , Ying H, , Yong WH, , Black KL, & Wheeler CJ: Vaccination with tumor lysate-pulsed dendritic cells elicits antigen-specific, cytotoxic T-cells in patients with malignant glioma. Cancer Res 64:49734979, 2004

    • Search Google Scholar
    • Export Citation
  • 50

    Yu JS, , Wheeler CJ, , Zeltzer PM, , Ying H, , Finger DN, & Lee PK, : Vaccination of malignant glioma patients with peptide-pulsed dendritic cells elicits systemic cytotoxicity and intracranial T-cell infiltration. Cancer Res 61:842847, 2001

    • Search Google Scholar
    • Export Citation
  • 51

    Zou JP, , Morford LA, , Chougnet C, , Dix AR, , Brooks AG, & Torres N, : Human glioma-induced immunosuppression involves soluble factor(s) that alters monocyte cytokine profile and surface markers. J Immunol 162:48824892, 1999

    • Search Google Scholar
    • Export Citation

Metrics

All Time Past Year Past 30 Days
Abstract Views 368 277 18
Full Text Views 176 11 1
PDF Downloads 63 7 1
EPUB Downloads 0 0 0