Cytotoxicity of cytotoxic T lymphcytes induced by keratin 19-sensitized dendritic cells against MCF-7 cells in vitro

Abstract

Abstract: Objective To investigate the cytotoxicity of cytotoxic T lymphcytes (CTL) induced by keratin 19 (K19)-sensitized dendritic cells (DC) against MCF-7 cells in vitro. Methods DC isolated from peripheral blood mononuclear cells were cultured in vitro and sensitized by K19 peptide to generate specific CTL. The phenotypes and intracytoplasm cytokine of DC were analyzed by flow cytometry. Mixed leukocyte responses were evaluated by ³H-TdR incorporation assay. The T cells were generated by DC pulsed with K19 antigen and T cell cytotoxicity was measured by ⁵¹Cr release assay. Results The DCs derived from peripheral blood mononuclear cells expressed high levels of CD40, CD86, CD80 and CD83. Mixed leukocyte responses induced by the DCs pulsed with K19 was stronger than that induced by naive DCs (P<0.01). The cytotoxicity rate of CTL induced by the sensitized DCs was higher than that of CTL induced by naive DCs (P<0.01). The cytotoxity activity was enhanced by increasing the effector/target ratio (P<0.01). Conclusion After sensitization with K19 antigen, the DCs can stimulate T cell proliferation and induce cytotoxicity activity against MCF-7 cells. Increased effector/target ratio may enhance the cytotoxity activity. Sensitized DCs possess the potential for amplification in immunotherapy of carcinoma.

CLC Number:

LI Ming-fang, LUO Rong-cheng, YOU Chang-xuan, TU Xiao-yu. Cytotoxicity of cytotoxic T lymphcytes induced by keratin 19-sensitized dendritic cells against MCF-7 cells in vitro[J]. Journal of Southern Medical University, 2005, 25(10): 1247-1250.

References

[1] Rosenblatt J, Kufe D, Avigan D. Dendritic cell fusion vaccines for cancer immunotherapy [J]. Expert Opin Biol Ther, 2005, 5 (5):703-15.
[2] Hayashi T, Nakao K, Nagayama Y, et al. Vaccination with dendritic cells pulsed with apoptotic cells elicits effective antitumor immunity in murine hepatoma models[J]. Int J Oncol, 2005, 26(5): 1313-9.
[3] Okano F, Merad M, Furumoto K, et al. In vivo manipulation of dendritic cells overcomes tolerance to unmodified tumor-associated self antigens and induces potent antitumor immunity [J]. J Immunol, 2005, 174(5): 2645-52.
[4] 尤长宣,罗荣城,苏瑾,等.腺相关病毒介导乳腺癌BA46基因转移的实验研究[J].中华微生物与免疫学杂志,2004,24(9):724-7.You CX, Luo RC, Su J, et al. Adeno-associated virus mediate the breast cancer BA46 gene transfer into dendritic cells [J]. Chin J Microbiol Immunol, 2004, 24(9): 724-7.
[5] Liu Y, You CX, Luo RC, et al. Use and specificity of breast cancer antigen/milk protein BA46 for generating anti-self-cytotoxic T lymphocytes by recombinated virus-based gene loading of dendritic cells[J]. Cancer Gene Ther, 2004, 12(1): 1-9.
[6] Zitvoge IL, Mayordomo J, Tjandrawan T, et al. Therapy of murine tumors with tumor peptide pulsed dendritic cells: dependence on T cells, B7 co-stimulation, and T help cell 1 associated cytokines[J]. J Exp Med, 1996, 183(1): 87-97.
[7] Nakamura I, Kajino K, Bamba H, et al. Phenotypic stability of mature dendritic cells tuned by TLR or CD40 to control the efficiency of cytotoxic T cell priming [J]. Microbiol Immunol, 2004, 48 (3):211-9.
[8] Colombo MP, Trinchieri G. Interleukin-12 in anti-tumor immunity and immunotherapy [J]. Cytokine Growth Factor Rev, 2002, 13(1):155-68.
[9] Heystek HC, Mudde GC, Ohler R, et al. Granulocyte-macrophage colony-stimulating factor (GM-CSF) has opposing effects on the capacity of monocytes versus monocyte-derived dendritic cells to stimulate the antigen-specific proliferation of a human T cell clone [J]. Clin Exp Immunol, 2000, 120(3): 440-7.
[10] Lutz MB, Schnare M, Menges M, et al. Differential functions of IL-4 receptor types Ⅰ and Ⅱ for dendritic cell maturation and IL-12 production and their dependency on GM-CSF [J]. J Immunol, 2002,169(7): 3574-80.
[11] Wan Y, Lu L, Bramson JL, et al. Dendritic cell-derived IL-12 is not required for the generation of cytotoxic, IFN-gamma-secreting, CD8 (+) CTL in vivo[J]. J Immunol, 2001, 167(9): 5027-33.
[12] Yang S, Vervaert CE, Burch J Jr, et al. Murine dendritic cells transfected with human GP100 elicit both antigen-specific CD8 (+) and CD4(+) T-cell responses and are more effective than DNA vaccines at generating anti-tumor immunity [J]. Int J Cancer, 1999, 83(4):532-40.