nach oben

Erschienen in:

01.11.2008 | Original Article

Immunotherapy of neuroblastoma by an Interleukin-21-secreting cell vaccine involves survivin as antigen

verfasst von: Michela Croce, Raffaella Meazza, Anna M. Orengo, Marina Fabbi, Martina Borghi, Domenico Ribatti, Beatrice Nico, Barbara Carlini, Vito Pistoia, Maria Valeria Corrias, Silvano Ferrini

Erschienen in: Cancer Immunology, Immunotherapy | Ausgabe 11/2008

Einloggen, um Zugang zu erhalten

Abstract

Aim

IL-21 is the most recently identified member of the IL-2 cytokine family. Here we studied the therapeutic efficacy of IL-21-gene-modified cells (Neuro2a/IL-21) in a syngeneic metastatic neuroblastoma (NB) model.

Materials and methods

Neuro2a/IL-21 cells were tested as subcutaneous (sc) vaccine both in prophylactic and therapeutic settings. Depletion studies, cytotoxicity assay and immunohistochemical analyses were carried out to evaluate the mechanisms involved in tumor rejection.

Results

When injected sc in syngeneic A/J mice viable Neuro2a/IL-21 cells were rejected and induced resistance to a subsequent iv challenge with Neuro2a parental cells (Neuro2a/pc), suggesting the involvement of an immune response. More importantly, in mice bearing Neuro2a/pc micrometastases, a single sc injection of Neuro2a/IL-21 cells significantly increased the mean tumor-free survival of treated animals (43 vs. 22 days) and cured 14% of them. The administration of two or three doses of Neuro2a/IL-21 cell vaccine further increased the mean survival time to 54 and 75 days, and the cure rate to 27 and 33%, respectively, whereas the use of unmodified Neuro2a or mock-transfected cells had no effect. In vivo cell subset depletion and a Winn-assay indicated the involvement of CD8 + CTLs. Immunohistochemical analysis indicated a reduction of CD31+ and VEGFR2+ microvessels in late metastases from therapeutically vaccinated mice. A role of survivin as antigen was suggested by in vitro assays using survivin-synthetic CTL-epitopes.

Conclusions

Our present data indicate that IL-21-secreting NB cells are effective as therapeutic vaccine in mice bearing metastatic NB, through a specific CTL response involving survivin as antigen, and suggest a potential interest for IL-21 in NB immuno-gene therapy.

Nur mit Berechtigung zugänglich

Adida C, Berrebi D, Peuchmaur M, Reyes-Mugica M, Altieri DC (1998) Anti-apoptosis gene, survivin, and prognosis of neuroblastoma. Lancet 351:882–883PubMedCrossRef

Airoldi I, Meazza R, Croce M, Di Carlo E, Piazza T, Cocco C, D’Antuono T, Pistoia V, Ferrini S, Corrias MV (2004) Low-dose interferon-gamma-producing human neuroblastoma cells show reduced proliferation and delayed tumorigenicity. Br J Cancer 90:2210–8PubMed

Beyer M, Schultze JL (2006) Regulatory T cells in cancer. Blood 108:804–811PubMedCrossRef

Brady J, Hayakawa Y, Smyth MJ, Nutt SL (2004) IL-21 induces the functional maturation of murine NK cells. J Immunol 172:2048–2058PubMed

Brandt K, Bulfone-Paus S, Foster DC, Ruckert R (2003) Interleukin-21 inhibits dendritic cell activation and maturation. Blood 102:4090–4098PubMedCrossRef

Casciano I, De Ambrosis A, Croce M, Pagnan G, Di Vinci A, Allemanni G, Banelli B, Ponzoni M, Romani M, Ferrini S (2004) Expression of the caspase-8 gene in neuroblastoma cells is regulated through an essential interferon-sensitive response element (ISRE). Cell Death Differ 11:131–134PubMedCrossRef

Comes A, Rosso O, Orengo AM, Di Carlo E, Sorrentino C, Meazza R, Piazza T, Valzasina B, Nanni P, Colombo MP, Ferrini S (2006) CD25+ regulatory T cell depletion augments immunotherapy of micrometastases by an IL-21-secreting cellular vaccine. J Immunol 176:1750–1758PubMed

Coughlin CM, Vance BA, Grupp SA, Vonderheide RH (2004) RNA-transfected CD40-activated B cells induce functional T-cell responses against viral and tumor antigen targets: implications for pediatric immunotherapy. Blood 103:2046–2054PubMedCrossRef

Croce M, Meazza R, Orengo AM, Radic L, De Giovanni B, Gambini C, Carlini B, Pistoia V, Mortara L, Accolla RS, Corrias MV, Ferrini S (2005) Sequential immunogene therapy with interleukin-12- and interleukin-15-engineered neuroblastoma cells cures metastatic disease in syngeneic mice. Clin Cancer Res 11:735–742PubMed

10.

De Bernardi B, Nicolas B, Boni L, Indolfi P, Carli M, Cordero Di Montezemolo L, Donfrancesco A, Pession A, Provenzi M, di Cataldo A, Rizzo A, Tonini GP, Dallorso S, Conte M, Gambini C, Garaventa A, Bonetti F, Zanazzo A, D’Angelo P, Bruzzi P, Italian Co-Operative Group for Neuroblastoma (2003) Disseminated neuroblastoma in children older than one year at diagnosis: comparable results with three consecutive high-dose protocols adopted by the Italian Co-operative Group for Neuroblastoma. J Clin Oncol 21:1592–1601PubMedCrossRef

11.

Di Carlo E, Comes A, Orengo AM, Rosso O, Meazza R, Musiani P, Colombo MP, Ferrini S (2004) IL-21 induces tumor rejection by specific CTL and IFN-gamma-dependent CXC chemokines in syngeneic mice. J Immunol 172:1540–1547PubMed

12.

Di Carlo E, De Totero D, Piazza T, Fabbi M, Ferrini S (2007) Role of IL-21 in immune-regulation and tumor immunotherapy. Cancer Immunol Immunother 56:1323–1334PubMedCrossRef

13.

Hoffmann D, Bayer W, Grunwald T, Wildner O (2007) Intratumoral expression of respiratory syncytial virus fusion protein in combination with cytokines encoded by adenoviral vectors as in situ tumor vaccine for colorectal cancer. Mol Cancer Ther 6:1942–1950PubMedCrossRef

14.

Huang AY, Gulden PH, Woods AS, Thomas MC, Tong CD, Wang W, Engelhard VH, Pasternack G, Cotter R, Hunt D, Pardoll DM, Jaffee EM (1996) The immunodominant major histocompatibility complex class I-restricted antigen of a murine colon tumor derives from an endogenous retroviral gene product. Proc Natl Acad Sci USA 93:9730–9735PubMedCrossRef

15.

Islam A, Kageyama H, Takada N, Kawamoto T, Takayasu H, Isogai E, Ohira M, Hashizume K, Kobayashi H, Kaneko Y, Nakagawara A (2000) High expression of Survivin, mapped to 17q25, is significantly associated with poor prognostic factors and promotes cell survival in human neuroblastoma. Oncogene 19:617–623PubMedCrossRef

16.

Kasaian MT, Whitters MJ, Carter LL, Lowe LD, Jussif JM, Deng B, Johnson KA, Witek JS, Senices M, Konz RF, Wurster AL, Donaldson DD, Collins M, Young DA, Grusby MJ (2002) IL-21 limits NK cell responses and promotes antigen-specific T cell activation: a mediator of the transition from innate to adaptive immunity. Immunity 16:559–569PubMedCrossRef

17.

King C, Ilic A, Koelsch K, Sarvetnick N (2004) Homeostatic expansion of T cells during immune insufficiency generates autoimmunity. Cell 117:265–277PubMedCrossRef

18.

Kowalczyk A, Wierzbicki A, Gil M, Bambach B, Kaneko Y, Rokita H, Repasky E, Fenstermaker R, Brecher M, Ciesielski M, Kozbor D (2007) Induction of protective immune responses against NXS2 neuroblastoma challenge in mice by immunotherapy with GD2 mimotope vaccine and IL-15 and IL-21 gene delivery. Cancer Immunol Immunother 56:1443–1458PubMedCrossRef

19.

Lode HN, Xiang R, Dreier T, Varki NM, Gillies SD, Reisfeld RA (1998) Natural killer cell-mediated eradication of neuroblastoma metastases to bone marrow by targeted interleukin-2 therapy. Blood 91:1706–1715PubMed

20.

Ma HL, Whitters MJ, Konz RF, Senices M, Young DA, Grusby MJ, Collins M, Dunussi-Joannopoulos K (2003) IL-21 activates both innate and adaptive immunity to generate potent antitumor responses that require perforin but are independent of IFN-gamma. J Immunol 171:608–615PubMed

21.

Maris JM, Matthay KK (1999) Molecular biology of neuroblastoma. J Clin Oncol 17:2264–2279PubMed

22.

Morandi F, Chiesa S, Bocca P, Millo E, Salis A, Solari M, Pistoia V, Prigione I (2006) Tumor mRNA-transfected dendritic cells stimulate the generation of CTL that recognize neuroblastoma-associated antigens and kill tumor cells: immunotherapeutic implications. Neoplasia 8:833–842PubMedCrossRef

23.

Moroz A, Eppolito C, Li Q, Tao J, Clegg CH, Shrikant PA (2004) IL-21 enhances and sustains CD8+ T cell responses to achieve durable tumor immunity: comparative evaluation of IL-2, IL-15, and IL-21. J Immunol 173:900–909PubMed

24.

Nakano H, Kishida T, Asada H, Shin-Ya M, Shinomiya T, Imanishi J, Shimada T, Nakai S, Takeuchi M, Hisa Y, Mazda O (2006) Interleukin-21 triggers both cellular and humoral immune responses leading to therapeutic antitumor effects against head and neck squamous cell carcinoma. J Gene Med 8:90–99PubMedCrossRef

25.

Nielsen M, Lundegaard C, Lund O, Kesmir C (2005) The role of the proteasome in generating cytotoxic T cell epitopes: Insights obtained from improved predictions of proteasomal cleavage. Immunogenetics 57:33–41PubMedCrossRef

26.

Ozaki K, Spolski R, Feng CG, Qi CF, Cheng J, Sher A, Morse HC 3rd, Liu C, Schwartzberg PL, Leonard WJ (2002) A critical role for IL-21 in regulating immunoglobulin production. Science 298:1630–1634PubMedCrossRef

27.

Parker KC, Bednarek MA, Coligan JE (1994) Scheme for ranking potential HLA-A2 binding peptides based on independent binding of individual peptide side-chains. J Immunol 152:163–175PubMed

28.

Parrish-Novak J, Dillon SR, Nelson A, Hammond A, Sprecher C, Gross JA, Johnston J, Madden K, Xu W, West J, Schrader S, Burkhead S, Heipel M, Brandt C, Kuijper JL, Kramer J, Conklin D, Presnell SR, Berry J, Shiota F, Bort S, Hambly K, Mudri S, Clegg C, Moore M, Grant FJ, Lofton-Day C, Gilbert T, Rayond F, Ching A, Yao L, Smith D, Webster P, Whitmore T, Maurer M, Kaushansky K, Holly RD, Foster D (2000) Interleukin-21 and its receptor are involved in NK cell expansion and regulation of lymphocyte function. Nature 408:57–63PubMedCrossRef

29.

Pertl U, Wodrich H, Ruehlmann JM, Gillies SD, Lode HN, Reisfeld RA (2003) Immunotherapy with a posttranscriptionally modified DNA vaccine induces complete protection against metastatic neuroblastoma. Blood 101:649–654PubMedCrossRef

30.

Ribatti D, Nico B, Pezzolo A, Vacca A, Meazza R, Cinti R, Carlini B, Parodi F, Pistoia V, Corrias MV (2006) Angiogenesis in a human neuroblastoma xenograft model: mechanisms and inhibition by tumour-derived interferon-gamma. Br J Cancer 94:1845–1852PubMedCrossRef

31.

Siegel S, Wagner A, Schmitz N, Zeis M (2003) Induction of antitumour immunity using survivin peptide-pulsed dendritic cells in a murine lymphoma model. Br J Haematol 122:911–914PubMedCrossRef

32.

Ugai S, Shimozato O, Kawamura K, Wang YQ, Yamaguchi T, Saisho H, Sakiyama S, Tagawa M (2003) Expression of the interleukin-21 gene in murine colon carcinoma cells generates systemic immunity in the inoculated hosts. Cancer Gene Ther 10:187–192PubMedCrossRef

33.

van Leeuwen EM, Gamadia LE, Baars PA, Remmerswaal EB, ten Berge IJ, van Lier RA (2002) Proliferation requirements of cytomegalovirus-specific, effector-type human CD8+ T cells. J Immunol 169:5838–5843PubMed

34.

Wang G, Tschoi M, Spolski R, Lou Y, Ozaki K, Feng C, Kim G, Leonard WJ, Hwu P (2003) In vivo antitumor activity of interleukin 21 mediated by natural killer cells. Cancer Res 63:9016–9022PubMed

35.

Wurster AL, Rodgers VL, Satoskar AR, Whitters MJ, Young DA, Collins M, Grusby MJ (2002) Interleukin 21 is a T helper (Th) cell 2 cytokine that specifically inhibits the differentiation of naïve Th cells into interferon gamma-producing Th1 cells. J Exp Med 196:969–977PubMedCrossRef

36.

Wysocki PJ, Karczewska-Dzionk A, Mackiewicz-Wysocka M, Mackiewicz A (2004) Human cancer gene therapy with cytokine gene-modified cells. Expert Opin Biol Ther 4:1595–1607PubMedCrossRef

37.

Xiang R, Mizutani N, Luo Y, Chiodoni C, Zhou H, Mizutani M, Ba Y, Becker JC, Reisfeld RA (2005) A DNA vaccine targeting survivin combines apoptosis with suppression of angiogenesis in lung tumor eradication. Cancer Res 65:553–561PubMed

Titel: Immunotherapy of neuroblastoma by an Interleukin-21-secreting cell vaccine involves survivin as antigen
verfasst von: Michela Croce
Raffaella Meazza
Anna M. Orengo
Marina Fabbi
Martina Borghi
Domenico Ribatti
Beatrice Nico
Barbara Carlini
Vito Pistoia
Maria Valeria Corrias
Silvano Ferrini
Publikationsdatum: 01.11.2008
Verlag: Springer-Verlag
Erschienen in: Cancer Immunology, Immunotherapy / Ausgabe 11/2008
Print ISSN: 0340-7004
Elektronische ISSN: 1432-0851
DOI: https://doi.org/10.1007/s00262-008-0496-3

Springer Medizin

Immunotherapy of neuroblastoma by an Interleukin-21-secreting cell vaccine involves survivin as antigen

Abstract