Top

Published in:

01-01-2011 | Preclinical study

High immunogenic potential of p53 mRNA-transfected dendritic cells in patients with primary breast cancer

Authors: Özcan Met, Eva Balslev, Henrik Flyger, Inge Marie Svane

Published in: Breast Cancer Research and Treatment | Issue 2/2011

Abstract

As pre-existent immunity might be a reflection of an emerging anticancer response, the demonstration of spontaneous T-cell responses against tumor associated antigens (TAAs) in cancer patients may be beneficial before clinical development of dendritic cell (DC)-based cancer vaccines, because it will help to identify likely responders to TAAs among patients who qualify and may benefit from this form of immune therapy. This study aimed to determine pre-existent T-cell reactivity against the tumor suppressor protein p53 in breast cancer patients (BCP) at the time point of primary diagnosis. After a short-term stimulation with autologous wt p53 mRNA-transfected DCs, IFN-γ enzyme-linked immunosorbent spot (ELISPOT) analysis revealed p53-reactive T cells in the peripheral blood of more than 40% (15 of 36) of the tested patients. Both CD4⁺ and CD8⁺ p53-specific T cells secreted IFN-γ after stimulation with p53-transfected DCs. Interestingly, more than 72% (13 of 18) of patients with high p53 (p53^high) expression in tumors were able to mount a p53-specific IFN-γ T-cell response, in contrast to only 10% (1 of 10) of healthy donors and 11% (2 of 18) of patients with low or absent p53 (p53^low) expression in tumors. Furthermore, significantly higher secretion of IL-2 was detected in peripheral blood mononuclear cells after stimulation with p53-transfected DCs from patients with p53^high tumor expression compared to patients with p53^low tumor expression, whereas secretion of IL-10 was predominant in the latter group. The high frequency of spontaneous wt p53-reactive T cells detected in the peripheral blood of primary BCP with accumulation of p53 in tumor provides a rationale to consider DCs transfected with mRNA encoding wt p53 for clinical investigation in these patients.

Dunn GP, Bruce AT, Ikeda H, Old LJ, Schreiber RD (2002) Cancer immunoediting: from immunosurveillance to tumor escape. Nat Immunol 3:991–998CrossRefPubMed

van der Bruggen P, Zhang Y, Chaux P et al (2002) Tumor-specific shared antigenic peptides recognized by human T cells. Immunol Rev 188:51–64CrossRef

Tacken PJ, de Vries IJ, Torensma R, Figdor CG (2007) Dendritic-cell immunotherapy: from ex vivo loading to in vivo targeting. Nat Rev Immunol 7:790–802CrossRefPubMed

Van Tendeloo VF, Ponsaerts P, Berneman ZN (2007) mRNA-based gene transfer as a tool for gene and cell therapy. Curr Opin Mol Ther 9:423–431PubMed

Van Tendeloo VF, Ponsaerts P, Lardon F et al (2001) Highly efficient gene delivery by mRNA electroporation in human hematopoietic cells: superiority to lipofection and passive pulsing of mRNA and to electroporation of plasmid cDNA for tumor antigen loading of dendritic cells. Blood 98:49–56CrossRefPubMed

Kessler JH, Melief CJ (2007) Identification of T-cell epitopes for cancer immunotherapy. Leukemia 21:1859–1874CrossRefPubMed

Theobald M, Biggs J, Dittmer D, Levine AJ, Sherman LA (1995) Targeting p53 as a general tumor antigen. Proc Natl Acad Sci USA 92:11993–11997CrossRefPubMed

DeLeo AB, Whiteside TL (2008) Development of multi-epitope vaccines targeting wild-type sequence p53 peptides. Expert Rev Vaccines 7:1031–1040CrossRefPubMed

Disis ML, Knutson KL, Schiffman K, Rinn K, McNeel DG (2000) Pre-existent immunity to the HER-2/neu oncogenic protein in patients with HER-2/neu overexpressing breast and ovarian cancer. Breast Cancer Res Treat 62:245–252CrossRefPubMed

10.

Nagorsen D, Scheibenbogen C, Schaller G et al (2003) Differences in T-cell immunity toward tumor-associated antigens in colorectal cancer and breast cancer patients. Int J Cancer 105:221–225CrossRefPubMed

11.

Rentzsch C, Kayser S, Stumm S et al (2003) Evaluation of pre-existent immunity in patients with primary breast cancer: molecular and cellular assays to quantify antigen-specific T lymphocytes in peripheral blood mononuclear cells. Clin Cancer Res 9:4376–4386PubMed

12.

Met O, Eriksen J, Svane IM (2008) Studies on mRNA electroporation of immature and mature dendritic cells: effects on their immunogenic potential. Mol Biotechnol 40:151–160CrossRefPubMed

13.

Pedersen AE, Thorn M, Gad M et al (2005) Phenotypic and functional characterization of clinical grade dendritic cells generated from patients with advanced breast cancer for therapeutic vaccination. Scand J Immunol 61:147–156CrossRefPubMed

14.

Garcia-Tunon I, Ricote M, Ruiz A et al (2006) Cell cycle control related proteins (p53, p21, and Rb) and transforming growth factor beta (TGFbeta) in benign and carcinomatous (in situ and infiltrating) human breast: implications in malignant transformations. Cancer Invest 24:119–125CrossRefPubMed

15.

McCutcheon M, Wehner N, Wensky A et al (1997) A sensitive ELISPOT assay to detect low-frequency human T lymphocytes. J Immunol Methods 210:149–166CrossRefPubMed

16.

Svane IM, Pedersen AE, Johansen JS et al (2007) Vaccination with p53 peptide-pulsed dendritic cells is associated with disease stabilization in patients with p53 expressing advanced breast cancer; monitoring of serum YKL-40 and IL-6 as response biomarkers. Cancer Immunol Immunother 56:1485–1499CrossRefPubMed

17.

Goodell V, Salazar LG, Urban N et al (2006) Antibody immunity to the p53 oncogenic protein is a prognostic indicator in ovarian cancer. J Clin Oncol 24:762–768CrossRefPubMed

18.

Valmori D, Scheibenbogen C, Dutoit V et al (2002) Circulating Tumor-reactive CD8(+) T cells in melanoma patients contain a CD45RA(+)CCR7(−) effector subset exerting ex vivo tumor-specific cytolytic activity. Cancer Res 62:1743–1750PubMed

19.

Karanikas V, Colau D, Baurain JF et al (2001) High frequency of cytolytic T lymphocytes directed against a tumor-specific mutated antigen detectable with HLA tetramers in the blood of a lung carcinoma patient with long survival. Cancer Res 61:3718–3724PubMed

20.

Hoffmann TK, Donnenberg AD, Finkelstein SD et al (2002) Frequencies of tetramer + T cells specific for the wild-type sequence p53(264–272) peptide in the circulation of patients with head and neck cancer. Cancer Res 62:3521–3529PubMed

21.

Chikamatsu K, Sakakura K, Takahashi G et al (2009) CD4+ T cell responses to HLA-DP5-restricted wild-type sequence p53 peptides in patients with head and neck cancer. Cancer Immunol Immunother 58:1441–1448CrossRefPubMed

22.

van der Burg SH, de Cock K, Menon AG et al (2001) Long lasting p53-specific T cell memory responses in the absence of anti-p53 antibodies in patients with resected primary colorectal cancer. Eur J Immunol 31:146–155CrossRefPubMed

23.

Sotiropoulou PA, Perez SA, Voelter V et al (2003) Natural CD8+ T-cell responses against MHC class I epitopes of the HER-2/neu oncoprotein in patients with epithelial tumors. Cancer Immunol Immunother 52:771–779CrossRefPubMed

24.

Lewis JD, Reilly BD, Bright RK (2003) Tumor-associated antigens: from discovery to immunity. Int Rev Immunol 22:81–112CrossRefPubMed

25.

Gnjatic S, Cai Z, Viguier M et al (1998) Accumulation of the p53 protein allows recognition by human CTL of a wild-type p53 epitope presented by breast carcinomas and melanomas. J Immunol 160:328–333PubMed

26.

Vierboom MP, Zwaveling S, Bos GMJ et al (2000) High steady-state levels of p53 are not a prerequisite for tumor eradication by wild-type p53-specific cytotoxic T lymphocytes. Cancer Res 60:5508–5513PubMed

27.

Inokuma M, dela Rosa C, Schmitt C et al (2007) Functional T cell responses to tumor antigens in breast cancer patients have a distinct phenotype and cytokine signature. J Immunol 179:2627–2633PubMed

28.

Sommerfeldt N, Schutz F, Sohn C et al (2006) The shaping of a polyvalent and highly individual T-cell repertoire in the bone marrow of breast cancer patients. Cancer Res 66:8258–8265CrossRefPubMed

29.

Nishikawa H, Kato T, Tanida K et al (2003) CD4+ CD25+ T cells responding to serologically defined autoantigens suppress antitumor immune responses. Proc Natl Acad Sci USA 100:10902–10906CrossRefPubMed

30.

Okada K, Komuta K, Hashimoto S et al (2000) Frequency of apoptosis of tumor-infiltrating lymphocytes induced by fas counterattack in human colorectal carcinoma and its correlation with prognosis. Clin Cancer Res 6:3560–3564PubMed

31.

Kurnick JT, Ramirez-Montagut T, Boyle LA et al (2001) A novel autocrine pathway of tumor escape from immune recognition: melanoma cell lines produce a soluble protein that diminishes expression of the gene encoding the melanocyte lineage melan-A/MART-1 antigen through down-modulation of its promoter. J Immunol 167:1204–1211PubMed

32.

Nikitina EY, Clark JI, Van Beynen J et al (2001) Dendritic cells transduced with full-length wild-type p53 generate antitumor cytotoxic T lymphocytes from peripheral blood of cancer patients. Clin Cancer Res 7:127–135PubMed

33.

Humrich J, Jenne L (2003) Viral vectors for dendritic cell-based immunotherapy. Curr Top Microbiol Immunol 276:241–259PubMed

34.

Reuschenbach M, von Knebel DM, Wentzensen N (2009) A systematic review of humoral immune responses against tumor antigens. Cancer Immunol Immunother 8:1535–1544CrossRef

35.

Vujanovic L, Ranieri E, Gambotto A et al (2006) IL-12p70 and IL-18 gene-modified dendritic cells loaded with tumor antigen-derived peptides or recombinant protein effectively stimulate specific Type-1 CD4+ T-cell responses from normal donors and melanoma patients in vitro. Cancer Gene Ther 13:798–805CrossRefPubMed

36.

Minkis K, Kavanagh DG, Alter G et al (2008) Type 2 Bias of T cells expanded from the blood of melanoma patients switched to type 1 by IL-12p70 mRNA-transfected dendritic cells. Cancer Res 68:9441–9450CrossRefPubMed

37.

Hung K, Hayashi R, Lafond-Walker A et al (1998) The central role of CD4(+) T cells in the antitumor immune response. J Exp Med 188:2357–2368CrossRefPubMed

38.

Pardoll DM, Topalian SL (1998) The role of CD4+ T cell responses in antitumor immunity. Curr Opin Immunol 10:588–594CrossRefPubMed

39.

Gilboa E, Vieweg J (2004) Cancer immunotherapy with mRNA-transfected dendritic cells. Immunol Rev 199:251–263CrossRefPubMed

40.

Muller MR, Grunebach F, Nencioni A, Brossart P (2003) Transfection of dendritic cells with RNA induces CD4- and CD8-mediated T cell immunity against breast carcinomas and reveals the immunodominance of presented T cell epitopes. J Immunol 170:5892–5896PubMed

41.

Kyte JA, Kvalheim G, Aamdal S, Saeboe-Larssen S, Gaudernack G (2005) Preclinical full-scale evaluation of dendritic cells transfected with autologous tumor-mRNA for melanoma vaccination. Cancer Gene Ther 12:579–591CrossRefPubMed

42.

Nimmerjahn F, Milosevic S, Behrends U et al (2003) Major histocompatibility complex class II-restricted presentation of a cytosolic antigen by autophagy. Eur J Immunol 33:1250–1259CrossRefPubMed

43.

Kalady MF, Onaitis MW, Padilla KM et al (2002) Enhanced dendritic cell antigen presentation in RNA-based immunotherapy. J Surg Res 105:17–24CrossRefPubMed

44.

Nair SK, Boczkowski D, Morse M et al (1998) Induction of primary carcinoembryonic antigen (CEA)-specific cytotoxic T lymphocytes in vitro using human dendritic cells transfected with RNA. Nat Biotechnol 16:364–369CrossRefPubMed

45.

Schaft N, Dorrie J, Thumann P et al (2005) Generation of an optimized polyvalent monocyte-derived dendritic cell vaccine by transfecting defined RNAs after rather than before maturation. J Immunol 174:3087–3097PubMed

46.

Javorovic M, Pohla H, Frankenberger B, Wolfel T, Schendel DJ (2005) RNA transfer by electroporation into mature dendritic cells leading to reactivation of effector-memory cytotoxic T lymphocytes: a quantitative analysis. Mol Ther 12:734–743CrossRefPubMed

47.

Wang RF, Wang HY (2002) Enhancement of antitumor immunity by prolonging antigen presentation on dendritic cells. Nat Biotechnol 20:149–154CrossRefPubMed

48.

He Y, Zhang J, Mi Z, Robbins P, Falo LD Jr (2005) Immunization with lentiviral vector-transduced dendritic cells induces strong and long-lasting T cell responses and therapeutic immunity. J Immunol 174:3808–3817PubMed

49.

Ludewig B, McCoy K, Pericin M et al (2001) Rapid peptide turnover and inefficient presentation of exogenous antigen critically limit the activation of self-reactive CTL by dendritic cells. J Immunol 166:3678–3687PubMed

50.

Bachmann MF, Beerli RR, Agnellini P et al (2006) Long-lived memory CD8+ T cells are programmed by prolonged antigen exposure and low levels of cellular activation. Eur J Immunol 36:842–854CrossRefPubMed

51.

Busch DH, Kerksiek KM, Pamer EG (2000) Differing roles of inflammation and antigen in T cell proliferation and memory generation. J Immunol 164:4063–4070PubMed

Title: High immunogenic potential of p53 mRNA-transfected dendritic cells in patients with primary breast cancer
Authors: Özcan Met
Eva Balslev
Henrik Flyger
Inge Marie Svane
Publication date: 01-01-2011
Publisher: Springer US
Published in: Breast Cancer Research and Treatment / Issue 2/2011
Print ISSN: 0167-6806
Electronic ISSN: 1573-7217
DOI: https://doi.org/10.1007/s10549-010-0844-9

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 2/2011

Lack of association between ATM C.1066-6T > G mutation and breast cancer risk: a meta-analysis of 8,831 cases and 4,957 controls

The β2-adrenergic receptor and Her2 comprise a positive feedback loop in human breast cancer cells

A high prevalence of BRCA1 mutations among breast cancer patients from the Bahamas

The association between two polymorphisms in pre-miRNAs and breast cancer risk: a meta-analysis

Targeting mutant p53 protein and the tumor vasculature: an effective combination therapy for advanced breast tumors

Vegetable intake is associated with reduced breast cancer recurrence in tamoxifen users: a secondary analysis from the Women’s Healthy Eating and Living Study

Keynote webinar | Spotlight on antibody–drug conjugates in cancer