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01-06-2008 | Original Article

Allogeneic tumor lysate can serve as both antigen source and protein supplementation for dendritic cell culture

Authors: Peter Dubsky, Hubert Hayden, Monika Sachet, Thomas Bachleitner-Hofmann, Michaela Hassler, Roswitha Pfragner, Michael Gnant, Anton Stift, Josef Friedl

Published in: Cancer Immunology, Immunotherapy | Issue 6/2008

Abstract

Background

Recent preclinical and clinical evidence suggests the use of allogeneic tumor as a source of antigen for DC-based immunotherapy against cancer. We hypothesized that addition of allogeneic tumor lysate to monocyte-derived DC culture could serve a dual purpose: (1) antigen source and (2) protein supplementation of DC culture media. Protein supplementation whether of known origin (human serum/plasma, fetal bovine serum, human serum albumin) or undeclared origin (“serum-free” media) is a source of variability and bias. We addressed the question whether protein supplementation can be omitted in the presence of allogeneic tumor lysate.

Materials and methods

Human DC cultured in the presence of lysate from medullary thyroid carcinoma (MTC) cell line SHER-I (TuLy-DC) and DC pulsed with the same lysate but cultured in the presence of FBS (FBS-DC) were assessed for morphology, phenotype, maturation and functional properties.

Results

In comparison of FBS-DC/TuLy-DC no significant differences in morphology, phenotype and maturation could be detected. Both culture conditions produced CD1a^high, CD14^low DC with high expression of costimulatory molecules and CD83 upon stimulation. TuLy-DC gave significantly better yields and produced more IL12p70. DC showed high (allo)stimulatory capacity toward T-cells. TuLy-DC induced more intracellular IFNγ in CD8+T-cells of vaccinated MTC patients. Both types of DC induced killing of SHER-I after short in vitro restimulation.

Summary and conclusion

Tumor lysate from SHER-I can substitute for further protein supplementation in DC culture. Allogeneic tumor lysates should be taken into consideration as both source of antigen and protein supplementation in monocyte-derived DC culture.

Saito H, Frleta D, Dubsky P, Palucka AK (2006) Dendritic cell-based vaccination against cancer. Hematol Oncol Clin North Am 20:689–710PubMedCrossRef

Banchereau J, Palucka AK, Dhodapkar M, Burkeholder S, Taquet N, Rolland A, Taquet S, Coquery S, Wittkowski KM, Bhardwaj N, Pineiro L, Steinman R, Fay J (2001) Immune and clinical responses in patients with metastatic melanoma to CD34(+) progenitor-derived dendritic cell vaccine. Cancer Res 61:6451–6458PubMed

Caux C, Vanbervliet B, Massacrier C, Dubois B, Dezutter-Dambuyant C, Schmitt D, Banchereau J (1995) Characterization of human CD34+ derived dendritic/Langerhans cells (D-Lc). Adv Exp Med Biol 378:1–5PubMed

Fong L, Hou Y, Rivas A, Benike C, Yuen A, Fisher GA, Davis MM, Engleman EG (2001) Altered peptide ligand vaccination with Flt3 ligand expanded dendritic cells for tumor immunotherapy. Proc Natl Acad Sci U S A 98:8809–8814PubMedCrossRef

Romani N, Gruner S, Brang D, Kampgen E, Lenz A, Trockenbacher B, Konwalinka G, Fritsch PO, Steinman RM, Schuler G (1994) Proliferating dendritic cell progenitors in human blood. J Exp Med 180:83–93PubMedCrossRef

Sallusto F, Lanzavecchia A (1994) Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha. J Exp Med 179:1109–1118PubMedCrossRef

Dubsky P, Ueno H, Piqueras B, Connolly J, Banchereau J, Palucka AK (2005) Human dendritic cell subsets for vaccination. J Clin Immunol 25:551–572PubMedCrossRef

Royer PJ, Tanguy-Royer S, Ebstein F, Sapede C, Simon T, Barbieux I, Oger R, Gregoire M (2006) Culture medium and protein supplementation in the generation and maturation of dendritic cells. Scand J Immunol 63:401–409PubMedCrossRef

Kadri N, Potiron N, Ouary M, Jegou D, Gouin E, Bach JM, Lieubeau B (2007) Fetal calf serum-primed dendritic cells induce a strong anti-fetal calf serum immune response and diabetes protection in the non-obese diabetic mouse. Immunol Lett 108:129–136 (Epub 2006 Dec 18)PubMedCrossRef

10.

Mackensen A, Drager R, Schlesier M, Mertelsmann R, Lindemann A (2000) Presence of IgE antibodies to bovine serum albumin in a patient developing anaphylaxis after vaccination with human peptide-pulsed dendritic cells. Cancer Immunol Immunother 49:152–156PubMedCrossRef

11.

Ostler T, Ehl S (2002) A cautionary note on experimental artefacts induced by fetal calf serum in a viral model of pulmonary eosinophilia. J Immunol Methods 268:211–218PubMedCrossRef

12.

Russo V, Tanzarella S, Dalerba P, Rigatti D, Rovere P, Villa A, Bordignon C, Traversari C (2000) Dendritic cells acquire the MAGE-3 human tumor antigen from apoptotic cells and induce a class I-restricted T cell response. Proc Natl Acad Sci USA 97:2185–2190PubMedCrossRef

13.

Mahdian R, Kokhaei P, Najar HM, Derkow K, Choudhury A, Mellstedt H (2006) Dendritic cells, pulsed with lysate of allogeneic tumor cells, are capable of stimulating MHC-restricted antigen-specific antitumor T cells. Med Oncol 23:273–282PubMedCrossRef

14.

Palucka AK, Ueno H, Connolly J, Kerneis-Norvell F, Blanck JP, Johnston DA, Fay J, Banchereau J (2006) Dendritic cells loaded with killed allogeneic melanoma cells can induce objective clinical responses and MART-1 specific CD8+ T-cell immunity. J Immunother 29:545–557PubMedCrossRef

15.

Vilella R, Benitez D, Mila J, Lozano M, Vilana R, Pomes J, Tomas X, Costa J, Vilalta A, Malvehy J, Puig S, Mellado B, Marti R, Castel T (2004) Pilot study of treatment of biochemotherapy-refractory stage IV melanoma patients with autologous dendritic cells pulsed with a heterologous melanoma cell line lysate. Cancer Immunol Immunother 53:651–658PubMedCrossRef

16.

Salcedo M, Bercovici N, Taylor R, Vereecken P, Massicard S, Duriau D, Vernel-Pauillac F, Boyer A, Baron-Bodo V, Mallard E, Bartholeyns J, Goxe B, Latour N, Leroy S, Prigent D, Martiat P, Sales F, Laporte M, Bruyns C, Romet-Lemonne JL, Abastado JP, Lehmann F, Velu T (2006) Vaccination of melanoma patients using dendritic cells loaded with an allogeneic tumor cell lysate. Cancer Immunol Immunother 55:819–829PubMedCrossRef

17.

Escobar A, Lopez M, Serrano A, Ramirez M, Perez C, Aguirre A, Gonzalez R, Alfaro J, Larrondo M, Fodor M, Ferrada C, Salazar-Onfray F (2005) Dendritic cell immunizations alone or combined with low doses of interleukin-2 induce specific immune responses in melanoma patients. Clin Exp Immunol 142:555–568PubMed

18.

Holtl L, Ramoner R, Zelle-Rieser C, Gander H, Putz T, Papesh C, Nussbaumer W, Falkensammer C, Bartsch G, Thurnher M (2005) Allogeneic dendritic cell vaccination against metastatic renal cell carcinoma with or without cyclophosphamide. Cancer Immunol Immunother 54:663–670PubMedCrossRef

19.

Pandha HS, John RJ, Hutchinson J, James N, Whelan M, Corbishley C, Dalgleish AG (2004) Dendritic cell immunotherapy for urological cancers using cryopreserved allogeneic tumour lysate-pulsed cells: a phase I/II study. BJU Int 94:412–418PubMedCrossRef

20.

Holtl L, Zelle-Rieser C, Gander H, Papesh C, Ramoner R, Bartsch G, Rogatsch H, Barsoum AL, Coggin JH Jr, Thurnher M (2002) Immunotherapy of metastatic renal cell carcinoma with tumor lysate-pulsed autologous dendritic cells. Clin Cancer Res 8:3369–3376PubMed

21.

Stift A, Friedl J, Dubsky P, Bachleitner-Hofmann T, Benkoe T, Brostjan C, Jakesz R, Gnant M (2003) In vivo induction of dendritic cell-mediated cytotoxicity against allogeneic pancreatic carcinoma cells. Int J Oncol 22:651–656PubMed

22.

Bachleitner-Hofmann T, Strohschneider M, Krieger P, Sachet M, Dubsky P, Hayden H, Schoppmann SF, Pfragner R, Gnant M, Friedl J, Stift A (2006) Heat shock treatment of tumor lysate-pulsed dendritic cells enhances their capacity to elicit antitumor T cell responses against medullary thyroid carcinoma. J Clin Endocrinol Metab 91:4571–4577PubMedCrossRef

23.

Stift A, Friedl J, Dubsky P, Bachleitner-Hofmann T, Schueller G, Zontsich T, Benkoe T, Radelbauer K, Brostjan C, Jakesz R, Gnant M (2003) Dendritic cell-based vaccination in solid cancer. J Clin Oncol 21:135–142PubMedCrossRef

24.

Stift A, Sachet M, Yagubian R, Bittermann C, Dubsky P, Brostjan C, Pfragner R, Niederle B, Jakesz R, Gnant M, Friedl J (2004) Dendritic cell vaccination in medullary thyroid carcinoma. Clin Cancer Res 10:2944–2953PubMedCrossRef

25.

Kern F, Surel IP, Brock C, Freistedt B, Radtke H, Scheffold A, Blasczyk R, Reinke P, Schneider-Mergener J, Radbruch A, Walden P, Volk HD (1998) T-cell epitope mapping by flow cytometry. Nat Med 4:975–978PubMedCrossRef

26.

27.

Bachleitner-Hofmann T, Stift A, Friedl J, Pfragner R, Radelbauer K, Dubsky P, Schuller G, Benko T, Niederle B, Brostjan C, Jakesz R, Gnant M (2002) Stimulation of autologous antitumor T-cell responses against medullary thyroid carcinoma using tumor lysate-pulsed dendritic cells. J Clin Endocrinol Metab 87:1098–1104PubMedCrossRef

28.

Schott M, Feldkamp J, Klucken M, Kobbe G, Scherbaum WA, Seissler J (2002) Calcitonin-specific antitumor immunity in medullary thyroid carcinoma following dendritic cell vaccination. Cancer Immunol Immunother 51:663–668 (Epub 2002 Oct 29)PubMedCrossRef

29.

Duperrier K, Eljaafari A, Dezutter-Dambuyant C, Bardin C, Jacquet C, Yoneda K, Schmitt D, Gebuhrer L, Rigal D (2000) Distinct subsets of dendritic cells resembling dermal DCs can be generated in vitro from monocytes, in the presence of different serum supplements. J Immunol Methods 238:119–131PubMedCrossRef

30.

Anton D, Dabadghao S, Palucka K, Holm G, Yi Q (1998) Generation of dendritic cells from peripheral blood adherent cells in medium with human serum. Scand J Immunol 47:116–121PubMedCrossRef

31.

Pietschmann P, Stockl J, Draxler S, Majdic O, Knapp W (2000) Functional and phenotypic characteristics of dendritic cells generated in human plasma supplemented medium. Scand J Immunol 51:377–383PubMedCrossRef

32.

Tarte K, Fiol G, Rossi JF, Klein B (2000) Extensive characterization of dendritic cells generated in serum-free conditions: regulation of soluble antigen uptake, apoptotic tumor cell phagocytosis, chemotaxis and T cell activation during maturation in vitro. Leukemia 14:2182–2192PubMedCrossRef

33.

Araki H, Katayama N, Mitani H, Suzuki H, Nishikawa H, Masuya M, Ikuta Y, Hoshino N, Miyashita H, Nishii K, Minami N, Shiku H (2001) Efficient ex vivo generation of dendritic cells from CD14+ blood monocytes in the presence of human serum albumin for use in clinical vaccine trials. Br J Haematol 114:681–689PubMedCrossRef

34.

Dubsky PC, Friedl J, Stift A, Bachleitner-Hofmann T, Jakesz R, Gnant MF, Weigel G (2006) Inosine 5’-monophosphate dehydrogenase inhibition by mycophenolic acid impairs maturation and function of dendritic cells. Clin Chim Acta 364:139–147PubMedCrossRef

35.

Kalinski P, Hilkens CM, Wierenga EA, Kapsenberg ML (1999) T-cell priming by type-1 and type-2 polarized dendritic cells: the concept of a third signal. Immunol Today 20:561–567PubMedCrossRef

36.

Trinchieri G, Pflanz S, Kastelein RA (2003) The IL-12 family of heterodimeric cytokines: new players in the regulation of T cell responses. Immunity 19:641–644PubMedCrossRef

37.

Bui HH, Sidney J, Dinh K, Southwood S, Newman MJ, Sette A (2006) Predicting population coverage of T-cell epitope-based diagnostics and vaccines. BMC Bioinformatics 7:153PubMedCrossRef

Title: Allogeneic tumor lysate can serve as both antigen source and protein supplementation for dendritic cell culture
Authors: Peter Dubsky
Hubert Hayden
Monika Sachet
Thomas Bachleitner-Hofmann
Michaela Hassler
Roswitha Pfragner
Michael Gnant
Anton Stift
Josef Friedl
Publication date: 01-06-2008
Publisher: Springer-Verlag
Published in: Cancer Immunology, Immunotherapy / Issue 6/2008
Print ISSN: 0340-7004
Electronic ISSN: 1432-0851
DOI: https://doi.org/10.1007/s00262-007-0422-0

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Abstract

Background

Materials and methods

Results

Summary and conclusion

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