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PLOS ONE  2013 

Recombinant Mammaglobin A Adenovirus-Infected Dendritic Cells Induce Mammaglobin A-Specific CD8+ Cytotoxic T Lymphocytes against Breast Cancer Cells In Vitro

DOI: 10.1371/journal.pone.0063055

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Mammaglobin A (MGBA) is a novel breast cancer-associated antigen almost exclusively over-expressed in primary and metastatic human breast cancers, making it a potential therapeutic target for breast cancer. The development of dendritic cell (DC)-induced tumor antigen specific CD8+ cytotoxic T lymphocytes (CTLs) may hold promise in cancer immunotherapy. In this study we constructed recombinant replication-defective adenoviral (Ad) vectors encoding MGBA and evaluated their ability to trigger anti-tumor immunity in vitro. DCs were isolated from the human peripheral blood monocyte cells (PBMCs) of two HLA-A33+ healthy female volunteers, and infected with adenovirus carrying MGBA cDNA (Ad-MGBA). After that, the Ad-MGBA-infected DCs were used to stimulate CD8+ CTLs in vitro and the latter was used for co-culture with breast cancer cell lines. The data revealed that infection with Ad-MGBA improved DC maturation and up-regulated the expression of co-stimulatory molecules and the secretion of interleukin-12 (IL-12), but down-regulated interleukin-10 (IL-10) secretion from DCs. Ad-MGBA-infected DC-stimulated CD8+CTLs displayed the highest cytotoxicity towards HLA-A33+/MGBA+ breast cancer MDA-MB-415 cells compared with other CD8+CTL populations, and compared with the cytotoxicity towards HLA-A33?/MGBA+ breast cancer HBL-100 cells and HLA-A33?/MGBA? breast cancer MDA-MB 231 cells. In addition, Ad-MGBA-infected DC-stimulated CD8+ CTLs showed a high level of IFNγ secretion when stimulated with HLA-A33+/MGBA+ breast cancer MDA-MB-415 cells, but not when stimulated with HLA-A33?/MGBA+ HBL-100 and HLA-A33?/MGBA?MDA-MB-231 cells. In addition, killing of CD8+CTLs against breast cancer was in a major histocompability complex (MHC)-limited pattern. Finally, the data also determined the importance of TNF-α in activating DCs and T cells. These data together suggest that MGBA recombinant adenovirus-infected DCs could induce specific anti-tumor immunity against MGBA+ breast cancers, which could provide a novel strategy in the immunotherapy of breast cancer.


[1]  He DD, Wang CF, Cao LL, Zhou J, Huang Jun, et al. (2010) Epidemic Trend of Female Breast Cancer Incidence in Min hang District, Shanghai. China Cancer 13: 108–10.
[2]  Gilboa E (2007) DC-based cancer vaccines. J Clin Invest 117: 1195–203.
[3]  Peng W, Zhao G, Ma Y, Yu H, Wang X (2011) Dendritic cells transfected with PEG10 recombinant adenovirus elicit anti-tumor immune response in vitro and in vivo. Vaccine 29: 3501–6.
[4]  Noh YW, Jang YS, Ahn KJ, Lim YT, Chung BH (2011) Simultaneous in vivo tracking of dendritic cells and priming of an antigen-specific immune response. Biomaterials 32: 6254–63.
[5]  Wierecky J, Mueller M, Brossart P (2006) Dendritic cell-based cancer immunotherapy targeting MUC-1. Cancer Immunol Immunother 55: 63–7.
[6]  Ilias Basha H, Tiriveedhi V, Fleming TP, Gillanders WE, Mohanakumar T (2011) Identification of immunodominant HLA-B7-restricted CD8+ cytotoxic T cell epitopes derived from mammaglobin-A expressed on human breast cancers. Breast Cancer Res Treat 127: 81–9.
[7]  Manna PP, Jaramillo A, Majumder K, Campbell LG, Fleming TP, et al. (2003) Generation of CD8+ cytotoxic T lymphocytes against breast cancer cells by stimulation with mammaglobin-A-pulsed dendritic cells. Breast Cancer Res Treat 79: 133–6.
[8]  Jaramillo A, Narayanan K, Campbell LG, Benshoff ND, Lybarger L, et al. (2004) Recognition of HLA-A2-restricted mammaglobin-A-derived epitopesby CD8+ cytotoxic T lymphocytes from breast cancer patients. Breast Cancer Res Treat 88: 29–41.
[9]  Wintermeyer P, Gehring S, Eken A, Wands JR (2010) Generation of cellular immune responses to HCV NS5 protein through in vivo activation of dendritic cells. J Viral Hepat 17: 705–13.
[10]  Chen X, Yu Y, Pan Q, Tang Z, Han J, et al. (2008) Enhancement of cytotoxic T lymphocyte activity by dendritic cells loaded with Tat-protein transduction domain-fused hepatitis B virus core antigen. Acta Biochim Biophys Sin 40: 996–1004.
[11]  Chamoto K, Kosaka A, Tsuji T, Matsuzaki J, Sato T, et al. (2003) Critical role of the Th1/Tc1 circuit for the generation of tumor-specific CTL during tumor eradication in vivo by Th1-cell therapy. Cancer Sci 94: 924–8.
[12]  Yoon SH, Yun SO, Park JY, Won HY, Kim EK, et al. (2009) Selective addition of CXCR3+CCR4-CD4+ Th1 cells enhances generation of cytotoxic T cells by dendritic cells in vitro. Exp Mol Med 41: 161–70.
[13]  Miyazawa M, Iwahashi M, Ojima T, Katsuda M, Nakamura M, et al. (2011) Dendritic cells adenovirally-transduced with full-length mesothelin cDNA elicit mesothelin-specific cytotoxicity against pancreatic cancer cell lines in vitro. Cancer Lett 305: 32–9.
[14]  Garetto S, Sizzano F, Brusa D, Tizzani A, Malavasi F, et al. (2009) Binding of prostate-specific membrane antigen to dendritic cells: a critical step in vaccine preparation. Cytotherapy 11: 1090–100.
[15]  Smits EL, Anguille S, Cools N, Berneman ZN, Van Tendeloo VF (2009) Dendritic cellbased cancer gene therapy. Hum Gene Ther 20: 1106–18.
[16]  Watson MA, Fleming TP (1994) Isolation of differentially expressed sequence tags from human breast cancer. Cancer Res 54: 4598–602.
[17]  Watson MA, Fleming TP (1996) Mammaglobin, a mammary specific member of the uteroglobin gene family, is overexpressed in human breast cancer. Cancer Res 56: 860–5.
[18]  Fleming TP, Watson MA (2000) Mammaglobin, a breast-specific gene, and its utility as a marker for breast cancer. Ann N Y Acad Sci 923: 78–89.
[19]  Jin SJ, Huang Y, Liu GX, Sun XD, Tan Y, et al. (2010) hMAM mRNA expression and clinical significance. Bull Acad Mil Med Sci l34: 440–3.
[20]  Tian S, Liu Z, Donahue C, Noh HS, Falo LD Jr, et al. (2009) Transcriptional IL-15-Directed in vivo DC Targeting DNA Vaccine. Gene Ther 16: 1260–70.
[21]  Bernstein JL, Godbold JH, Raptis G, Watson MA, Levinson B, et al. (2005) Identification of Mammaglobin as a Novel Serum Marker for Breast Cancer. Clin Cancer Res 11: 6528–35.
[22]  Kim J, Jayaprakasha GK, Patil BS (2013) Limonoids and their anti-proliferative and anti-aromatase properties in human breast cancer cells. Food Funct 4(2) 258–65.
[23]  Ceballos MP, Zumoffen C, Massa E, Cipulli G, Funes CC, et al. (2011) Detection of mammaglogin A in blood from breast cancer patients, before and after treatment, using a one-tube nested PCR protocol. Association with the absence of tumor estrogen receptors. Clin Biochem 44: 1429–33.
[24]  Mikhitarian K, Martin RH, Ruppel MB, Gillanders WE, Hoda R, et al. (2008) Detection of mammaglobin mRNA in peripheral blood is associated with high grade breast cancer: interim results of a prospective cohort study. BMC Cancer 8: 55.
[25]  Al-Joudi FS, Kaid FA, Ishak I, Mohamed N, Osman K, et al. (2011) Expression of human mammaglobin and clinicopathologic correlations in breast cancer: The findings in Malaysia. Indian J Pathol Microbiol 54(2): 284–9.
[26]  Aline F, Brand D, Pierre J, Roingeard P, Séverine M, et al. (2009) Dendritic cells loaded with HIV-1 p24 proteins adsorbed on surfactant-free anionic PLA nanoparticles induce enhanced cellular immune responses against HIV-1 after vaccination. Vaccine 27: 5284–91.
[27]  Koch F, Stanzl U, Jennewein P, Janke K, Heufler C, et al. (1996) High level IL-12 production by murine dendritic cells up-regulation via MHC class II and CD40 molecules and downregulation by IL-4 and IL-10. J Exp Med 184: 741–6.
[28]  Young JW (1999) Dendritic cell: expansion and differentiation with hematopoietic growth factors. Curr Opin Hematol. 6(3): 135–44.
[29]  Micheva I, Thanopoulou E, Michalopoulou S, Karakantza M, Kouraklis- Symeonidis A, et al. (2004) Defective tumor necrosis factor alpha-induced maturation of monocyte-derived dendritic cells in patients with myelodysplastic syndromes. Clin Immunol 113(3): 310–7.
[30]  Chen B, Shi Y, Smith JD, Chol D, Geiger JD, et al. (1998) The role of tumor necrosis factor-a in modulating the quantity of peripheral blood-derived, cytokine driven human dendritic cells and its role in enhancing the quality of dendritic cell function in presenting soluble antigens to cd4+ t cells in vitro. Blood 91(12) 4652–61.
[31]  Hirata N, Yanagawa Y, Ogura H, Satoh M, Noguchi M, et al. (2011) The role of tumor necrosis factor-a for interleukin-10 production by murine dendritic cells. Cell immunol 266(2): 165–71.
[32]  Herrera OB, Brett S, Lechler RI (2002) Infection of mouse bone marrow- derived cells with recombinant adenovirus vectors leads to presentation of eccoded antigen by both MHC class I and class II molecules-potential benefits in vaccine design. Vaccine 21(3–4): 231–42.
[33]  Vujanovic L, Theresa L, Potter DM, Chu J, Ferrone S, et al. (2009) Regulation of antigen presentation machinery in human dendritic cells by recombinant adenovirus. Cancer Immunol Immunother 58(1): 121–33.
[34]  Xie LH, Sin FW, Cheng SC, Cheung YK, Chan KT, et al. (2008) Activation of cytotoxic T lymphocytes against CML28-bearing tumors by dendritic cells transduced with a recombinant adeno-associated virus encoding the CML28 gene. Cancer Immunol Immunother 57: 1029–38.
[35]  Ge C, Xing Y, Wang Q, Xiao W, Lu Y, et al. (2011) Improved efficacy of therapeutic vaccination with dendritic cells pulsed with tumor cell lysate against hepatocellular carcinoma by introduction of 2 tandem repeats of microbial HSP70 peptide epitope 407–426 and OK-432.Int Immunopharmacol. 11: 2200–7.
[36]  Hirschowitz EA, Weaver JD, Hidalgo GE, Doherty DE (2000) Murine dentritic cells infected with adenovirus vectors show signs of activation. Gene therapy 7: 1112–20.
[37]  Philpott NJ, Nociari M, Elkon KB, Falck-Pedersen E (2004) Adenovirus- induced maturation of dendritic cells through a PI3 kinase-mediated TNF-a induction pathway. Proc Natl Acad Sci U S A 101(16): 6200–5.
[38]  Morelli AE, Larregina AT, Ganster RW, Zahorchak AF, Plowey JM, et al. (2000) Recombinant adenovirus induces maturation of dendritic cells via an NF-KB-dependent pathway. J Virol 74(20): 9617–28.
[39]  Balwin AS Jr (1995) The NF- n B and In B proteins: new discoveries and insights. Annu Rev Immunol 14 649–83.
[40]  Wu YG, Wu GZ, Wang L, Zhang YY, Li Z, et al. (2010) Tumor cell lysate-pulsed dendritic cells induce a T cell response against colon cancer in vitro and in vivo. Med Oncol 27: 736–42.
[41]  Narayanan K, Jaramillo A, Benshoff ND, Campbell LG, Fleming TP, et al. (2004) Response of Established Human Breast Tumors to Vaccination with Mammaglobin-A cDNA. J Natl Cancer Inst 96: 1388–96.
[42]  Chen JH, Yu YS, Chen XH, Liu HH, Zang GQ, et al. (2012) Enhancement of CTLs induced by DCs loaded with ubiquitinated hepatitis B virus core antigen. World J Gastroenterol 18: 1319–27.
[43]  Li A, Xiong S, Lin Y, Liu R, Chu Y (2011) A High-Affinity T-Helper Epitope Enhances Peptide-Pulsed Dendritic Cell-Based Vaccine. DNA Cell Biol 30: 883–92.
[44]  Tiriveedhi V, Sarma NJ, Subramanian V, Fleming TP, Gillanders WE, et al. (2012) Identification of HLA-A24- restricted CD8(+) cytotoxic T-cell epitopes derived from mammaglobin-A, a human breast cancer-associated antigen. Hum Immunol 73: 11–6.
[45]  Wei CH, Uhlin M, Masucci MG, Levitsky V (2002) Tetramer binding and secretion of interferon-a in response to antigenic stimulation are compatible with a range of affinities of MHC:TCR interaction and distinct programs of cytotoxic T-lymphocyte activation. Hum Immunol 63: 821–33.


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