Laboratory Investigation

Transcription

1 KISEP J Korean Neurosurg Soc 34 : Laboratory Investigation Induction of Cellular Immune Response by Dendritic Cells Pulsed with Glioma Apoptotic Bodies In-Su Kim, M.D., 1 Jong-Tae Kim, M.D., 1 Hyun-Il Cho, Ph.D., 2 Young-Sup Park, M.D., 1 Moon-Chan Kim, M.D., 1 Dong-Sup Chung, M.D. 1 Departments of Neurosurgery, 1 Microbiology, 2 College of Medicine, The Catholic University of Korea, Seoul, Korea Objective : The aim of this study is to verify the hypothesis that human dendritic cells(dcs) can process antigens from glioma cell apoptotic bodies and induce antigen-specific effector T cells. Methods : DCs generated in the presence of granulocyte macrophage-colony stimulating factor(gm-csf) and interleukin-4(il-4) from peripheral blood mononuclear cells(pbmcs) of healthy donors with human leucocyte antigen(hla) A*0201 were cultured for 7 days. Glioma apoptotic bodies(gabs) from T98G glioblastoma cells following 18 hour-actinomycin D treatment were co-incubated with DCs for 3 days. CD8 T cells isolated from peripheral blood of same donors were cultured in media containing IL-2 and were stimulated by GAB-pulsed DCs three times at a weekly interval. The interferon-(ifn-), a cytokine related to cytotoxicity, concentrations of cell culture supernates were measured by enzyme immunoassay technique. Results : Induced DCs had DC's own phenotypic characteristics such as highly expressed major histocompatibility complex(mhc) class II, CD1a and CD86 molecules. They also had high endocytotic activity. Preteatment of T98G glioma cells with actinomycin D resulted in 53% of cells undergoing apoptosis. IFN-production of effector T cells stimulated by GAB-pulsed DCs was significantly higher than that of T cells stimulated by non-pulsed DCs. Conclusion : Naïve CD8 T cells can be activated by human GAB-pulsed DCs to become antigen-specific effector T cells. Using GABs as a antigen source may be a novel approach in future DC-based immunotherapeutic trials for malignant glioma. KEY WORDS : Dendritic cell Glioma apoptotic body Interferon- Antigen-specific effector T cell Immunotherapeutic trial. ï J Korean Neurosurg Soc 34

2 IS Kim, et al.

3 Induction of Immune Response J Korean Neurosurg Soc 34

4 IS Kim, et al. Table 1. Production of interferon- by T cells of 3 groups determined by immunoassay

5 Induction of Immune Response J Korean Neurosurg Soc 34

6 IS Kim, et al. Acknowledgement References 1. Albert ML, Sauter B, Bhardwaj N : Dendritic cells acquire antigen from apoptotic cells and induce class I-restricted CTLs. Nature 392 : 86-89, Ashley DM, Faiola B, Nair S, Hale LP, Bigner DD, Gilboa E : Bone marrow-generated dendritic cells pulsed with tumor extracts or tumor RNA induce antitumor immunity against central nervous system tumors. J Exp Med 186 : , Banchereau J, Steinman RM : Dendritic cells and the control of immunity. Nature 392 : , Celluzzi CM, Falo LD Jr : Physical interaction between dendritic cells and tumor cells results in an immunogen that induces protective and therapeutic tumor rejection. J Immunol 160 : , Celluzzi CM, Mayordomo JI, Storkus WJ, Lotze MT, Falo LD Jr : Peptide-pulsed dendritic cells induce antigen-specific CTL-mediated protective tumor immunity. J Exp Med 183 : , Chang JW, Peng M, Vaquerano JE, Zhou YM, Clinton RA, Hyun WC, et al : Induction of Th1 response by dendritic cells pulsed with autologous melanoma apoptotic bodies. Anticancer Res 20 : , Chen Z, Moyana T, Saxena A, Warrington R, Jia Z, Xiang J : Efficient antitumor immunity derived from maturation of dendritic cells that had phagocytosed apoptotic/necrotic tumor cells. Int J Cancer 93 : , Croft M, Carter L, Swain SL, Dutton RW : Generation of polarized antigen specific CD8 effector populations: reciprocal actions of interleukin (IL)-4 and IL-12 in promoting type 2 versus type 1 cytokine profiles. J Exp Med 180 : , Dobrzanski MJ, Reome JB, Dutton RW : Therapeutic effects of tumor-reactive type 1 and type 2 CD8+ T cell subpopulations in established pulmonary metastases. J Immunol 162 : , Henkart PA : Lymphocyte-mediated cytotoxicity: two pathways and multiple effector molecules. Immunity 1 : , Hishii M, Nitta T, Ishida H, Ebato M, Kurosu A, Yagita H, et al : Human glioma-derived interleukin-10 inhibits antitumor immune responses in vitro. Neurosurgery 37 : , Hiyama H, Reeves SA : Role for cyclin D1 in UVC-induced and p53-mediated apoptosis. Cell Death Differ 6 : , Hoffmann TK, Meidenbauer N, Dworacki G, Kanaya H, Whiteside TL : Generation of tumor-specific T lymphocytes by cross-priming with human dendritic cells ingesting apoptotic tumor cells. Cancer Res 60 : , Hunter CA, Reiner SL : Cytokines and T cells in host defense. Curr Opin Immunol 12 : , Kuppner MC, Hamou MF, Sawamura Y, Bodmer S, de Tribolet N : Inhibition of lymphocyte function by glioblastoma-derived transforming growth factor 2. J Neurosurg 71 : , Morford LA, Elliott LH, Carlson SL, Brooks WH, Roszman TL : T cell receptor-mediated signaling is defective in T cells obtained from patients with primary intracranial tumors. J Immunol 159 : , Mosmann TR, Li L, Sad S : Functions of CD8 T-cell subsets secreting different cytokine patterns. Semin Immunol 9 : 87-92, Narita Y, Asai A, Kuchino Y, Kirino T : Actinomycin D and staurosporine, potent apoptosis inducers in vitro, are potentially effective chemotherapeutic agents against glioblastoma multiforme. Cancer Chemother Pharmacol 45 : , Noble A, Macary PA, Kemeny DM : IFN- and IL-4 regulate the growth and differentiation of CD8+ T cells into subpopulations with distinct cytokine profiles. J Immunol 155 : , Park JT, Kim TY : Cisplatin-induced apoptosis by cisplatin in human glioblastoma cell line. J Korean Neurosurg Soc 31 : , Rubin J, Lotze M : Adoptive cellular immunotherapy of cancer in Mitchel M (ed) : Biological approaches to cancer treatment : biomodulation, New York : McGraw-Hill, Inc., 1993, pp Schnurr M, Scholz C, Rothenfusser S, Galambos P, Dauer M, Robe J, et al : Apoptotic pancreatic tumor cells are superior to cell lysates in promoting cross-priming of cytotoxic T cells and activate NK and T cells. Cancer Res 62 : , Trojan J, Johnson TR, Rudin SD, Ilan J, Tykocinski ML, Ilan J : Treatment and prevention of rat glioblastoma by immunogenic C6 cells expressing antisense IGF-I RNA. Science 259 : 94-97, Yu JS, Wheeler CJ, Zeltzer PM, Ying H, Finger DN, Lee PK, et al : Vaccination of malignant glioma patients with peptide-pulsed dendritic cells elicits systemic cytotoxicity and intracranial T-cell infiltration. Cancer Res 61 : , 2001