CpG ODN对肺癌和淋巴瘤细胞的作用研究
摘要
CpG ODN是含有CpG基序的寡聚脱氧核苷酸,能够激发机体产生Th1样免疫应答,在肿瘤治疗方面展现出广泛的应用前景。本研究采用自行设计的CpG ODN,以细胞计数及MTT掺入法在体外观察了其对肺癌细胞生长的抑制作用和对化疗药长春新碱(VCR)治疗恶性淋巴瘤的增效作用;以小鼠Lewis肺癌移植瘤模型在体内观察了其对肺癌的治疗作用。结果显示:B型CpG ODN可在体外明显抑制人肺癌细胞系A549细胞的增殖(P<0.05),在体内对小鼠Lewis肺癌移植瘤有抑制趋势,但不如C型CpG ODN的抑制趋势明显。将B型CpG ODN用于人B细胞淋巴瘤Raj(i伯基特淋巴瘤)细胞时表现出明显刺激细胞增殖的活性,其与化疗药VCR联用时则能增强VCR对Raji细胞的杀伤作用,但对人T淋巴细胞白血病Jurkat细胞无此作用。说明B型CpG ODN可能作为B型淋巴细胞来源的肿瘤的化疗增效剂,C型CpG ODN具有治疗肺癌的潜在价值。
This study includes two parts, the first part is the in vitro and in vivo study of CpG ODN alone in the treatment of lung cancer; the second part is the in vitro study of CpG ODN on human B cell lymphoma and acute T cell leukemia with or without VCR (vincristine).
Part 1. Effects of CpG ODNs on A549 cell and Lewis Lung carcinoma xenotransplant in mice
Lung cancer is a leading cause of death in all cancers world-wide, non-small cell lung cancer (NSCLC) takes the most part (about 80%) of it. The cause of lung cancer is still not clear, but datas have indicated that smoking cigarettes is the most common cause, the more years a person smokes, the greater the risk. Besides that, it also has relations with air pollution, individual’s immune states and hereditary factors.
Currently, treatments of non-small cell lung cancer are maily focused on surgery, chemotherapy and radiotherapy, combined with biological therapy, targeted therapy and Chinese therapy. Although surgery is the first choice, most NSCLC patients (>70%) lost the opportunity who are diagnosed in the late stage. For these patients, chemotherapy could prolong survival and improve life quality. But as a result of the non-selectivity of chemo drugs, patients who are frail and elderly could not tolerate the side effects. For these reasons, many biological therapy and targeted therapy are developed in recent years.
Targeted therapy of NSCLC concludes Endothelial growth factor receptor (EGFR) inhibitors, Angiogenesis inhibitor, Signal transduction inhibitors and Matrix metalloproteinase inhibitors. Currently these targeted therapy drugs in clinical trials are mostly used in combination with chemotherapy to enhance the effect of chemotherapeutic agents. However, most of these targeted therapy drugs only have a single target, which means they can not fully activate the immune system to inhibit the tumor growth. In addition, a number of targeted therapy drugs are still in clinical trials, and the effects are not very satisfactory.
Synthetic non-methylated CpG-oligodeoxynucleotides (CpG ODNs) are agonists of Toll-like receptor 9 (TLR9) that have been shown to be potent activators of immune system, which can provoke various immune cells and induce the production of a wide variety of T helper-1 (Th1)-promoting cytokines, such as interleukin 12 (IL-12), interferon-γ(IFN-γ), tumor necrosis factor-α(TNF-α) and IL-6. As a strong immunostimulatory agent, CpG ODN is able to protect the body from bacterial, viral and parasitic infections, and the administration of CpG ODN alone is also capable of triggering potent anti-tumor immune responses against various experimental tumors, for example, lymphoma, leukemia, glioma, breast cancer, colon carcinoma, fibrosarcoma, melanoma, lung cancer, uterine cervix cancer, cervical cancer, neuroblastoma. CpG ODN also enhances the anti-tumor efficacy of monoclonal antibodies or cancer vaccines when used as immune adjuvants in animal tumor models.
In this study, two CpG ODNs that were designed by ourselves were tested : B-type CpG ODN (ODN 01) and C-type CpG ODN (ODN 02). We detected the direct inhibitory effects of these two ODNs on human lung cancer cell A549 in vitro and mice Lewis lung cancer in vivo.
1. Direct effect of CpG ODNs on A549 cell
We adopted cell counting and MTT assay to detect the direct inhibitory effect of ODN 01 on A549 cell incubated in 96 well plate, conODN (without CG motif) as negative control.
The result shows that ODN 01 has inhibitory effect on A549 cell growth compared to medium and conODN group though there is some difference between cell count and MTT assay result. The inhibitory effect is related to cell density and the time of ODN 01 administration. The higher cell density put in the well, the stronger inhibitory effect of ODN 01 exhibits. Especially when A549 cells grew up to fill up the well, means the cells could contact with each other, the inhibitory effect of ODN 01 is obviously detected. As this result, we speculate that ODN 01 probably induced A549 cell to have the nature of contact inhibition again.
2. Effects of CpG ODNs in mice bearing Lewis lung cancer xenotransplant
Female C57BL/6 mice were inoculated subcutaneously right posterior on the back with Lewis lung cancer cell 2×106 per mouse at day 0. When the transplanted tumor could be seen at about day 10, we considered tumors are successfully transplanted. Mice bearing Lewis lung cancer were randomly divided into 3 groups: PBS, ODN 01 and ODN 02. Administration with ODN 25μg per mouse began at day 10, subcutaneously injected at right groin every other day, 6 times all together. By measuring the tumor volume, we found the growth rate of tumors on mice of group ODN 02 was the lowest compared with ODN 01 and PBS group. We could draw the conclusion that ODN 02 had inhibitory effect on Lewis lung cancer bearing mice.
Part two: Effects of CpG ODN on lymphoma/leukemia cells
Non Hodgkin’s lymphoma (NHL) is the most common disease in lymphatic system malignant tumor, B cell derived NHL takes about 85 percents. The morbidity and mortality trends are increasing year by year. NHL is sensitive to the first time chemotherapy, but recurrence and drug resistance is common. The therapy effect will significantly decrease when these patients receive secondary chemotherapy, so NHL is considered as difficult to cure.
Burkitt’s lymphoma (BL) is a kind of high invasion NHL that often develops outside the lymph node or develops to acute leukemia. BL is most happens on African children, more males than females. In our country it often happens on children and youths.
Vincristine (VCR) is a alkaloid extracted from oleander plant Vinca. Because of the good anti-tumor activity, it is widely use in acute lymphocytic leukemia, Hodgkin’s and non- Hodgkin’s lymphoma, and also breast cancer, bronchiogenic cancer, soft tissue sarcoma and neuroblastoma. The anti-tumor mechanism of VCR is to binding to microtubulin, inhibiting microtubule polymerization, and spindle can not be formed so that cells stop at metaphase of cell division. Besides that, VCR interferes protein synthesis and RNA polymerase so that it can kill G1 phase cell as well.
The target of VCR is the cells in the dividing phase, and B-type CpG ODN can stimulate the proliferation of B cells, which means taking the cells into dividing phase. On account of this two mechanism, we tried to use ODN 01 combined with VCR on Raji cells to enhance the anti-tumor effect of VCR.
1. Effects of CpG ODN on human malignant B cell lymphoma Raji cell with or without VCR
First we investigated the effect of ODN 01 alone on Raji cells. Cell counting and MTT assay result showed that ODN 01, as a B-type CpG ODN, could significantly stimulate malignant B cell lymphoma proliferation compared to medium group.
Next, we investigated the effect of ODN 01 combined with VCR on Raji cells. We found a suitable concentration of VCR, under which VCR had almost no lethal effect on Raji cells (Invalid maximum dose , 10ng/mL). We use ODN 01 combined with this concentration of VCR administrated on Raji cells. By cell counting and MTT assay we found ODN 01+VCR (10ng/mL) had stronger inhibitory effect on Raji cells compared to VCR (10ng/mL) alone. According to these results, we can draw a conclusion that ODN 01 could enhance the anti-tumor activity of VCR.
2. Effects of CpG ODN on human acute T cell leukemia Jurkat cells with or without VCR
We used same methods as above to investigate the effect of CpG ODN on human T cell leukemia Jurkat cells. On one hand, neither stimulatory nor inhibitory effect can be seen of ODN 01 on Jurkat cells. On the other hand, Jurkat cell is not that sensitive to VCR as Raji cell. Consequently, there was no enhancement of VCR with ODN 01 can be seen when they administrated together on Jurkat cells.
The innovation of this study is the utilization of B-type CpG ODN that can stimulate malignant B cell proliferation as an enhancer of VCR on B-cell lymphoma/leukemia therapy.
This study includes two parts, the first part is the in vitro and in vivo study of CpG ODN alone in the treatment of lung cancer; the second part is the in vitro study of CpG ODN on human B cell lymphoma and acute T cell leukemia with or without VCR (vincristine).
Part 1. Effects of CpG ODNs on A549 cell and Lewis Lung carcinoma xenotransplant in mice
Lung cancer is a leading cause of death in all cancers world-wide, non-small cell lung cancer (NSCLC) takes the most part (about 80%) of it. The cause of lung cancer is still not clear, but datas have indicated that smoking cigarettes is the most common cause, the more years a person smokes, the greater the risk. Besides that, it also has relations with air pollution, individual’s immune states and hereditary factors.
Currently, treatments of non-small cell lung cancer are maily focused on surgery, chemotherapy and radiotherapy, combined with biological therapy, targeted therapy and Chinese therapy. Although surgery is the first choice, most NSCLC patients (>70%) lost the opportunity who are diagnosed in the late stage. For these patients, chemotherapy could prolong survival and improve life quality. But as a result of the non-selectivity of chemo drugs, patients who are frail and elderly could not tolerate the side effects. For these reasons, many biological therapy and targeted therapy are developed in recent years.
Targeted therapy of NSCLC concludes Endothelial growth factor receptor (EGFR) inhibitors, Angiogenesis inhibitor, Signal transduction inhibitors and Matrix metalloproteinase inhibitors. Currently these targeted therapy drugs in clinical trials are mostly used in combination with chemotherapy to enhance the effect of chemotherapeutic agents. However, most of these targeted therapy drugs only have a single target, which means they can not fully activate the immune system to inhibit the tumor growth. In addition, a number of targeted therapy drugs are still in clinical trials, and the effects are not very satisfactory.
Synthetic non-methylated CpG-oligodeoxynucleotides (CpG ODNs) are agonists of Toll-like receptor 9 (TLR9) that have been shown to be potent activators of immune system, which can provoke various immune cells and induce the production of a wide variety of T helper-1 (Th1)-promoting cytokines, such as interleukin 12 (IL-12), interferon-γ(IFN-γ), tumor necrosis factor-α(TNF-α) and IL-6. As a strong immunostimulatory agent, CpG ODN is able to protect the body from bacterial, viral and parasitic infections, and the administration of CpG ODN alone is also capable of triggering potent anti-tumor immune responses against various experimental tumors, for example, lymphoma, leukemia, glioma, breast cancer, colon carcinoma, fibrosarcoma, melanoma, lung cancer, uterine cervix cancer, cervical cancer, neuroblastoma. CpG ODN also enhances the anti-tumor efficacy of monoclonal antibodies or cancer vaccines when used as immune adjuvants in animal tumor models.
In this study, two CpG ODNs that were designed by ourselves were tested : B-type CpG ODN (ODN 01) and C-type CpG ODN (ODN 02). We detected the direct inhibitory effects of these two ODNs on human lung cancer cell A549 in vitro and mice Lewis lung cancer in vivo.
1. Direct effect of CpG ODNs on A549 cell
We adopted cell counting and MTT assay to detect the direct inhibitory effect of ODN 01 on A549 cell incubated in 96 well plate, conODN (without CG motif) as negative control.
The result shows that ODN 01 has inhibitory effect on A549 cell growth compared to medium and conODN group though there is some difference between cell count and MTT assay result. The inhibitory effect is related to cell density and the time of ODN 01 administration. The higher cell density put in the well, the stronger inhibitory effect of ODN 01 exhibits. Especially when A549 cells grew up to fill up the well, means the cells could contact with each other, the inhibitory effect of ODN 01 is obviously detected. As this result, we speculate that ODN 01 probably induced A549 cell to have the nature of contact inhibition again.
2. Effects of CpG ODNs in mice bearing Lewis lung cancer xenotransplant
Female C57BL/6 mice were inoculated subcutaneously right posterior on the back with Lewis lung cancer cell 2×106 per mouse at day 0. When the transplanted tumor could be seen at about day 10, we considered tumors are successfully transplanted. Mice bearing Lewis lung cancer were randomly divided into 3 groups: PBS, ODN 01 and ODN 02. Administration with ODN 25μg per mouse began at day 10, subcutaneously injected at right groin every other day, 6 times all together. By measuring the tumor volume, we found the growth rate of tumors on mice of group ODN 02 was the lowest compared with ODN 01 and PBS group. We could draw the conclusion that ODN 02 had inhibitory effect on Lewis lung cancer bearing mice.
Part two: Effects of CpG ODN on lymphoma/leukemia cells
Non Hodgkin’s lymphoma (NHL) is the most common disease in lymphatic system malignant tumor, B cell derived NHL takes about 85 percents. The morbidity and mortality trends are increasing year by year. NHL is sensitive to the first time chemotherapy, but recurrence and drug resistance is common. The therapy effect will significantly decrease when these patients receive secondary chemotherapy, so NHL is considered as difficult to cure.
Burkitt’s lymphoma (BL) is a kind of high invasion NHL that often develops outside the lymph node or develops to acute leukemia. BL is most happens on African children, more males than females. In our country it often happens on children and youths.
Vincristine (VCR) is a alkaloid extracted from oleander plant Vinca. Because of the good anti-tumor activity, it is widely use in acute lymphocytic leukemia, Hodgkin’s and non- Hodgkin’s lymphoma, and also breast cancer, bronchiogenic cancer, soft tissue sarcoma and neuroblastoma. The anti-tumor mechanism of VCR is to binding to microtubulin, inhibiting microtubule polymerization, and spindle can not be formed so that cells stop at metaphase of cell division. Besides that, VCR interferes protein synthesis and RNA polymerase so that it can kill G1 phase cell as well.
The target of VCR is the cells in the dividing phase, and B-type CpG ODN can stimulate the proliferation of B cells, which means taking the cells into dividing phase. On account of this two mechanism, we tried to use ODN 01 combined with VCR on Raji cells to enhance the anti-tumor effect of VCR.
1. Effects of CpG ODN on human malignant B cell lymphoma Raji cell with or without VCR
First we investigated the effect of ODN 01 alone on Raji cells. Cell counting and MTT assay result showed that ODN 01, as a B-type CpG ODN, could significantly stimulate malignant B cell lymphoma proliferation compared to medium group.
Next, we investigated the effect of ODN 01 combined with VCR on Raji cells. We found a suitable concentration of VCR, under which VCR had almost no lethal effect on Raji cells (Invalid maximum dose , 10ng/mL). We use ODN 01 combined with this concentration of VCR administrated on Raji cells. By cell counting and MTT assay we found ODN 01+VCR (10ng/mL) had stronger inhibitory effect on Raji cells compared to VCR (10ng/mL) alone. According to these results, we can draw a conclusion that ODN 01 could enhance the anti-tumor activity of VCR.
2. Effects of CpG ODN on human acute T cell leukemia Jurkat cells with or without VCR
We used same methods as above to investigate the effect of CpG ODN on human T cell leukemia Jurkat cells. On one hand, neither stimulatory nor inhibitory effect can be seen of ODN 01 on Jurkat cells. On the other hand, Jurkat cell is not that sensitive to VCR as Raji cell. Consequently, there was no enhancement of VCR with ODN 01 can be seen when they administrated together on Jurkat cells.
The innovation of this study is the utilization of B-type CpG ODN that can stimulate malignant B cell proliferation as an enhancer of VCR on B-cell lymphoma/leukemia therapy.
引文
[1] AKIRA S, TAKEDA K. Functions of Toll-like receptors: lessons from KO mice. C. R. Biologies. 2004; 327: 581–589
[2] AKIRA S, TAKEDA K. Toll-like receptor signalling. Nat Rev Immunol. 2004; 4(7): 499-511
[3] BABATZ J, ROLLIG C, LOBEL B, et al. Induction of cellular immune responses against carcinoembryonic antigen in patients with metastatic tumors after vaccination with altered peptide ligand-loaded dendritic cells. J. Cancer Immunol Immunother, 2005, 54(2): 1-9
[4] BALLAS ZK, RASMUSSEN WL, KRIEG AM. Induction of NK activity in murine and HUMAN cells by CpG motifs in oligodeoxynucleotides and bacterial DNA. J Immunol. 1996, 157(5): 1840-5.
[5] BALSARI A, TORTORETO M, BESUSSO D, PETRANGOLINI G, SFONDRINI L, MAGGI R, MENARD S, PRATESI G. Combination of a CpG-oligodeoxynucleotide and a topoisomerase I inhibitor in the therapy of human tumour xenografts. Eur J Cancer. 2004 May; 40(8): 1275-81.
[6] BAUER S, KIRSCHNINQ CJ , HACKER H. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. Proc Nal A cad Sci U S A , 2001, 98 (16): 9237-9242
[7] BENDIGS S, SALZER U, LIPFORD GB, et al. CpG-oligodeoxynucleotides co-stimulate primary T cells in the absence of antigen-presenting cells. Eur J Immunol, 1999, 29 (4): 1209-1218.
[8] BERNASCONI N.L., ONAI N. & LANZAVECCHIA A. A role for Toll-like receptors in acquired immunity:up-regulation of TLR9 by BCR triggering in naive cells and constitutive expression in memory B cells Blood ,2003,101, 4500–4504.
[9] BISSETT D, O′BYRNE KJ, VON PAWEL J, et al. PhaseⅢstudy of matrix metalloproteinase inhibitor prinomastat in non-small-cell lung cancer. J. Clin Oncol, 2005, 23 (4): 842-849.
[10] BRENNAN FR, BELLABY T, HELLIWELL SM, et al. Chimeric plant virus particles administered nasally or orally induce systemic and mucosal immune responses in mice. J Virol, 1999, 73(2): 930-938.
[11] BRUNNER C, SEIDERER J, SCHLAMP A, BIDLINGLNAIER M, EIGLER A, HAIMERL W, et al. Enhanced dendritic cell maturation by TNF-alpha or cytidine-phosphate-guanosine DNA drives T cell activation in vitro and therapeutic anti-tumor immune responses in vivo. J Immunol. 2000, 165: 6278-86.
[12] CARPENTIER A, LAIGLE-DONADEY F, ZOHAR S, CAPELLE L, BEHIN A, TIBI A, MARTIN-DUVERNEUIL N, SANSON M, LACOMBLEZ L, TAILLIBERT S, PUYBASSET L, VAN EFFENTERRE R, DELATTRE JY, CARPENTIER AF. Phase 1 trial of a CpG oligodeoxynucleotide for patients with recurrent glioblastoma. Neuro Oncol. 2006, 8(1): 60-6.
[13] CHACE JH, HOOKER NA, MILDENSTEIN KL, KRIEG AM, COWDERY JS. Bacterial DNA-induced NK cell IFN-gamma production is dependent on macrophage secretion of IL-12. Clin immunol Immunopathol. 1997, 84: 185-93.
[14] COLEY WB. The treatment of malignant tumors by repeated inoculations of erysipelas with a report of ten original cases. Am J Med Sci. 1893; 105: 487–511.
[15] DAVILA E, KENNEDY R, CELIS E. Generation of antitumor immunity by cytotoxic T lymphocyte epitope peptide vaccination, CpG-oligodeoxynucleotide adjuvant and CTLA-4 blockade. Cancer Res. 2003. 63(12): 3281-8.
[16] DECKER T, HIPP S, KREITMAN RJ, PASTAN I, PESCHEL C, LICHT T. Sensitization of B-cell chronic lymphocytic leukemia cells to recombinant immunotoxin by immunostimulatory phosphorothioate oligodeoxynucleotides. Blood. 2002. 99 (4): 1320-6.
[17] DROEMANN D, ALBRECHT D, GERDES J, ULMER AJ, BRANSCHEID D, VOLLMER E, DALHOFF K, ZABEL P, GOLDMANN T. Human lungcancer cells express functionally active Toll-like receptor 9. Respir Res. 2005 Jan 4; 6:1.
[18] DUNNE A, O’NEILL LAJ. The interleukin-1 receptor/toll-like receptor superfamily: signal transduction during inflammation and host defense. J Science’s STKE, February 25, 2003, 2003(171): re3-3
[19] EL ANDALOUSSI A, SONABEND AM, HAN Y, LESNIAK MS. Stimulation of TLR9 with CpG ODN enhances apoptosis of glioma and prolongs the survival of mice with experimental brain tumors. Glia. 2006 Nov 1; 54(6): 526-35.
[20] FADERL S, THOMAS DA, O′BRIEN S, et al. Experience with alemtuzumab plus rituximab in patients with relapsed and refractory lymphoid malignancies. Blood, 2003, 101: 3413-3415.
[21] FELDMAN E, KALAYCIO M, WEINER G, et al. Treatment of relapsed or refractory acute myeloid leukemia with humanized anti-CD33 monoclonal antibody HuM195. Leukemia, 2003, 17: 3142 318
[22] FRIEDBERG JW, KIM H, MCCAULEY M, HESSEL EM, SIMS P, FISHER DC, NADLER LM, COFFMAN RL, FREEDMAN AS. Combination immunotherapy with a CpG olgonucleotide (1018 ISS) and rituximab in patients with non-Hodgkin lymphoma: increased interferon-alpha/beta-inducible gene expression, without significant toxicity. Blood. 2005, 105(2): 489-95
[23] FUKUOKA M, YANO S, GIACCONE G, et al. Multi-institutional randomized phaseⅡtrial of gefinitib for previously treated patients with advanced non-small-cell lung cancer. J Clin Oncol, 2003, 21 (12): 2237.
[24] GATZEMEIER U, GROTH G, BRITTS C, et al. Randomized phaseⅡtrial of Gemcitabine -Cisplatin with or without trastuzumab in Her-2 positive non-small cell lung cancer. J. Ann Oncol, 2004, 15(1):19-27.
[25] GRIDELLI C, MAIONE P, DEL GAIZO F, et al. Sorafenib and sunitinib in the treatment of advanced non-small-cell lung cancer. Oncologist, 2007, 12(2): 191-200
[26] GROSSBARD ML, MULTANI PS, FREEDMAN AS, et al. A Phase II study of adjuvant therapy with anti-B4-blocked ricin after autologous bone marrow transplantation for patients with relapsed B-cell non-Hodgkin′s lymphoma. Clin Cancer Res, 1999, 5:2392-2398
[27] HALE G, SLAVIN S, GOLDMAN JM. Alemtuzumab (Campath-1H) for treatment of lymphoid malignancies in the age of nonmyeloablative conditioning? Bone Marrow Transplant, 2002, 30: 797-804.
[28] HARTMANN G, WEERATNA RD, BALLAS ZK, PAYETTE P, BLACKWELL S, SUPARTO I, RASMUSSEN WL, WALDSCHMIDT M, SAJUTHI D, PURCELL RH, DAVIS HL, KRIEG AM. Delineation of a CpG Phosphorothioate Oligodeoxynucleotide for Activating Primate Immune Responses In Vitro and In Vivo. J Immunol. 2000; 164: 1617–1624
[29] HE H, CRIPPEN TL, FARNELL MB, KOGUT MH. Identification of CpG oligodeoxynucleotide motifs that stimulate nitric oxide and cytokine production in avian macrophage and peripheral blood mononuclear cells. Dev Comp Immunol. 2003, 27: 621-7
[30] HERBST RS, GIACCONE G, SCHILLER JH, et al. Gefitinib in combination with paclitaxel and carboplatin in advanced non-small-cell lung cancer: a phaseⅢtrial- INTACT 2 . J Clin Oncol, 2004, 22 (5): 785.
[31] HERBST RS, PRAGERD, HERMAN R, et al. TRIBUTE: A phaseⅢtrial of erlotinib hydrochloride (OSI-774) combined with carboplatin and paclitaxel chemotherapy in advanced non-small–cell lung cancer. J Clin Oncol, 2005, 23 (25): 5892.
[32] HEYMACH J, WEST H, KERR R, et al. ZD6474 in combination with carbopatin and paclitaxel as first line treatment in patients with NSCLC: Results of the run- in phase of a two-part randomized phaseⅡstudy. J. Lung Cancer, 2005, 49(Supp 12): S247.
[33] HURWITZ H, HOLDEN SN, ECKHARDT SG, et al. Clinical evaluation of ZD6474, an orally active inhibitor of VEGF signaling, in patients with solid tumors. J. Proc Am Soc Clin Oncol, 2002, 21: 82a
[34] IHO S, YAMAMOTO T, TAKAHASHI T, YAMAMOTO S. Oligodeoxynucleotides containing palindrome sequences with internal 5’-CpG-3’act directly on human NK and activated T cells to induce IFN-gamma production in vitro. J Immunol. 1999, 163: 3642-52
[35] JAHRSDORFER B, JOX R, MUHLENHOFF L, TSCHOEP K, KRUG A, ROTHENFUSSER S, MEINHARDT G, EMMERICH B, ENDRES S, HARTMANN G. Modulation of malignant B cell activation and apoptosis by bcl-2 antisense ODN and immunostimulatory CpG ODN. J Leukoc Biol. 2002, 70: 83-92
[36] JAHRSDORFER B, MüHLENHOFF L, BLACKWELL SE, WAGNER M, POECK H, HARTMANN E, JOX R, GIESE T, EMMERICH B, ENDRES S, WEINER GJ, HARTMANN G. B-cell lymphomas differ in their responsiveness to CpG oligodeoxynucleotides. J. Clin Cancer Res. 2005 Feb 15; 11(4): 1490-9.
[37] JAHRSDORFER, B. et al. 2005. Immunostimulatory oligodeoxynucleotides induce apoptosis of B cell chronic lymphocytic leukemia cells. J. Leukoc. Biol. 77: 378-387.
[38] JOHNSON DH, FEHRNBACHER L, NOVOTNY WF, et al. Randomized phaseⅡtrial comparing bevacizumb plus carboplatin and paclitaxel with carboplatin and paclitaxel alone in previously untreated locally advanced or metastic non-small-cell lung cancer. J Clin Oncol, 2004 ,22 (11): 2184-2191.
[39] KADOWAKI N, ANTONENKO S, LIU YJ. Distinct CpG DNA and polyinosinic-polycytidylic acid double-stranded RNA, respectively, stimulate CD11c-type 2 dendritic cell precursors and CD11c+ dendritic cells to produce type I IFN. J Immunol.2001 Feb 15; 166(4): 2291-5.
[40] KAWARADA Y, GANSS R, GARBI N, SACHER T, AMOLD B, H?MMERLING GJ. NK+ and CD8+ T Cell-Mediated Eradication of Established Tumors by Peritumoral Injection of CpG-Containing Oligodeoxynucleotides J Innunol. 2001, 167(9): 5247-53
[41] KIM, Y, GIRARDI M, DUVIC M, KUZEL T, ROOK A, LINK B. TLR9agonist immunomodulator treatment of cutaneous T-cell lymphoma(CTCL) with CpG 7909. ASH Annual Meeting Abstracts 104. 2004, Abstract 743
[42] KLINMAN DM,YI AK, BEAUCAGE SL, CONOVER J,KREIG AM. CPG motifs Present in bacteria DNA rapidly induce lymphocytes to secrete interleukin 6,interleukin 12, and interferon gamma. Proc Natl Acad Sci USA. 1996, 93: 2879-83
[43] KOHLER G, MILSTEIN C. Continuous cultures of fused cells secreting antibody of predefined specificity. J Immunol, 2005, 174(5): 2453.
[44] KRIEG AM. Toll-like receptor9 (TLR9) agonists in the treatment of cancer. Oncogene. 2008, 27(2): 161-7
[45] KRIEG AM. Therapeutic potential of Toll-like receptor 9 activation. Nat Rev Drug Discov. 2006 Jun; 5(6): 471-84.
[46] KRIEG AM,YI AK,MATSON S,WALDSCHMIDT TJ,BISHOP GA,TEASDALE R,KORETZKY GA,KLINMAN DM.CpG motifs in bacterial DNA trigger direct B-cell activation.Nature.1995;374:546-549
[47] KRIS MG, NATALE RB, HERBST RS, et al. A phaseⅡtrial of ZD1839 (Iressa) in advanced non-small-cell lung cancer (NSCLC) patients who had failed platinum-and docetaxel-based regi mens (IDEAL-2). J. Pro Am Soc Clin Oncol, 2002, 21: 292.
[48] KOLUMAM GA, S THOMAS, L J THOMPSON, J SPRENT, K MURALI-KRISHNA. TypeⅠinterferons act directly on CD8 T cells to allow clonal expansion and memory formation in response to viral infection. J Exp Med. 2005, 202: 637-650
[49] LANGER C J, STEPHENSON P, THOR A, et al. Trastuzumab in the treatment of advanced non-small-cell lung cancer : Is there a role? Focus on Eastern Cooperative Oncology Group Study 2598. J. J Clin Oncol, 2004, 22 (7): 1180-1187.
[50] LATZ E, SCHOENEMEYER A, VISINTINA, FITZGERALD KA, MONKS BG, KNETTER CF, LIEN E, NILSEN NJ, ESPEVIK T, GOLENBOCK DT. TLR9 signals after translocating from the ER to CpG DNA in the lysosome.Nat lmmunol. 2004, 5(2): 190-8.
[51] LEIGHL NB, PAZ-ARES L, DOUILLARD JY, et al. Randomized phaseⅢstudy of matrix metalloproteinase inhibit or BMS-275291 in combination with paclitaxel and carboplatin in advanced non-small-cell lung cancer: National Cancer Institute of Canada-Clinical Trials Group Study BR. 18. J. Clin Oncol, 2005, 23 (12): 2831-2839
[52] LINK BK, BALLAS ZK. WEISDORF D, WOOLDRIDGE JE, BOSSLER AD, SHANNON M, RASMUSSEN WL, KRIEG AM, WEINER GJ. Oligodeoxynucleotide CPG7909 delivered as intravenous infusion demonstrates immunologic modulation in patients with previously treated non-Hodgkin lymphoma. J. Immunother. 2006, 29(5): 558-68
[53] MANEGOLD C, et al. Addition of PF-3512676 (CpG 7909) to a taxane/platinum regimen for first-line treatment of unresectable non-small cell lung cancer (NSCLC) improves objective response-phaseⅡclinical trial [abstract]. Eur J Cancer. 2005, 3: 326
[54] MARRACK, P., J. Kappler, and T. Mitchell. TypeⅠinterferons keep activated T cells alive. J. Exp. Med. 1999, 189: 521-530
[55] MASON KA, ARIGA H, NEAL R, et al. Targeting toll-like receptor 9 with CpG oligodeoxynucleotides enhances tumor response to fractionated radiotherapy. J. Clin Cancer Res, 2005, 11(1): 361-369
[56] MASON KA, NEAL R, HUNTER N, et al. CpG oligodeoxynucleotides are potent enhancers of radio- and chemo responses of murine tumors. J. Radio ther Oncol, 2006, 80 (2): 192-198
[57] MELLOR, A.L., et al. 2005. Cutting edge: CpG oligonucleotides induce splenic CD19+ dendritic cells to acquire potent indoleamine 2, 3-dioxygenase-dependent T cell regulatory functions via IFN Type 1 signaling. J. Immunol. 175:5601–5605.
[58] MILAS L, MASON KA, ARIGA H, HUNTER N, NEAL R, VALDECANAS D, KRIEG AM, WHISNANT JK. CpG oligodeoxynucleotide enhances tumor response to radiation. Cancer Res. 2004, 64(15): 5074-7
[59] MODELSKA A, DIETZSCHOLD B, SLEYSH N, et al. Immunization against rabies with plant-derived antigen. J Proc Natl Acad Sci USA, 1998, 95(5): 2481-2485.
[60] MOLENKAMP BG, VAN LEEUWEN PA, VAN DEN EERTWEGH AJ, SLUIJTER BJ, SCHEPER RJ, MEIJER S, DE GRUIJL TD. Immunomoeulation of the melanoma sentinel lymph node: a novel adjuvant therapeutic option. Immunobiology. 2006, 211(6-8): 651-61
[61] NATALE R, BODKIN D, GOVINDAN R, et al. A comparison of the antitumor efficacy of ZD6474 and gefitinib (Iressa) in patients with NSCLC: results of a randomized, double blind, phaseⅡstudy. Lung Cancer, 2005, 49 (2): s371
[62] NIETHAMMER AG, PRIMUS FJ, XIANG R, et al. An oral DNA vaccine against human carcinoembryonic antigen (CEA) prevents growth and dissemination of Lewis lung carcinoma in CEA transgenic mice. Vaccine, 2001,20(3-4): 421-429
[63] ONN A, CORREA AM, GILCREASE M, et al. Synchronous over-expression of epidermal growth factor receptor and HER2-neu protein is a predict or of poor outcome in patients with stage I non-small cell lung cancer. Clin Cancer Res, 2004, 10 (1) : 136-143
[64] ONO M, KUWANO M. Molecular mechanisms of epidermal growth factor receptor (EGFR) activation and response to gefitinib and other EGFR-targeting drugs. J. Clin Cancer Res, 2006, 12 (24): 7242-7251.
[65] PASHENKOV M, GOЁSSG, WAGNER C, H?RMANN M, JANDL T, MOSER A, BRITTEN CM, SMOLLE J, KOLLER S, MAUCH C, TANTCHEVA-POOR L, GRABBE S, LOQUAI C, ESSER S, FRANCKSON T, SCHNEEBERGER A, HAARMANN C, KRIEG AM, STINGL G, WAGNER SN. PhaseⅡtrial of a toll-like receptor 9-activating oligonueleotide in patients with metastatic melanoma. J. Clin Oncol. 2006, 24(36): 5716-24
[66] PICCALUGA P P, ARTINELL I G, RONDON IM, et al. Gemtuzumabozogamicin for relapsed and refractory acute myeloid leukemia and myeloid sarcomas. J Leuk Lymphoma, 2004, 45 (9): 1791-1795
[67] PILON-THOMAS S, LI W, BRIGGS JJ, DJEU J, MULéJJ, RIKER AI. lmmunostimulatory effects of CpG-ODN upon dendritic cell-based immunotherapy in a murine melanoma model. J Immunother (1997). 2006, 29(4): 381-7
[68] RUTZ M, METZGER J, GELLERT T, LUPPA P, LIPFORD GB, WAGNER H, BAUER S. Toll-like receptor 9 binds single-stranded CpG-DNA in a sequence-and pH-dependent manner. Eur J Immunol. 2004 Sep; 34(9): 2541-50.
[69] ROSELL R, GATZEMEIER U, BETTICHER DC, et al. PhaseⅢrandomized trial comparing paclitaxel/carboplatin with paclitaxel/cisplatin in patients with advanced non-small-cell Lung cancer: A cooperative multinational trial. J. Ann Oncol, 2002, 13(10): 1539-1549
[70] SCHATTENBERG D, SCHOTT M, REINDL G, KRUEGER T, TSCHOEPE D, FELDKAMP J, SCHERBAUM WA, SEISSLER J. Response of human monocyte-derived dendritic cells to immunostimulatory DNA. Eur J Immunol. 2000; 30(10): 2824-31.
[71] SCHILLER JH, HARRINGTON D, BELANI CP, et al. Comparison of four chemotherapy regimens for advanced non- small- cell lung cancer. J. N Engl J Med, 2002, 346(2): 92-98
[72] SHARMA S, KARAKOUSIS CP, TAKITA H, SHIN K, BROOKS SP. Intra-tumoral injection of CpG results in the inhibition of tumor growth in murine Colon-26 and B-16 tumors. Biotechnol Lett. 2003, 25(2): 149-53
[73] SHEPHERD FA, RODRIGUES PEREIRA J, CIULEANU T, et a1. Erlotinib in previously treated non-small-cell lung cancer. N Engl J Med, 2005, 353 (2): 123-132
[74] SHEPHERD FA, PEREIRA J, CIULEANU JE, et al. A randomized placebo-controlled trial of erlotinib in patients with advanced non-small-cell lung cancer (NSCLC) following failure of 1st line or 2nd line chemotherapy. ANational Cancer Institute of Canada Clinical Trials Group (NCICCTG) Trial. J. Proc Am Soc Clin Oncol, 2004, 23: Abs 7022
[75] SHODA LK, KEGERREIS KA, SUAREZ CE, MWANGI W, KNOWLES DP, BROWN WC. Immunostimulatory CpG-modified plasmid DNA enhances IL-12, TNF-alpha, and NO production by bovine macrophages. J Leukoc Biol. 2001, 70(l): 103-12
[76] SIEVERS E L, LARSON R A, STADTMAUER E A, et al. Efficacy and safety of gemtuzumab ozogamicin in patients with CD33-positive acute myeloid leukemia in first relapse. J Clin Oncol, 2001, 19 (13): 3244-3254
[77] SPEISER DE, LIéNARD D, RUFER N, RUBIO-GODOY V, RIMOLDI D, LEJEUNE F, KRIEG AM, CEROTTINI JC, ROMERO P. Rapid and strong human CD8+ T cell responses to wanccination with peptide, IFA, and CpG oligodeoxynucleotide 7909. J Clin Invest. 2005, 115(3):739-46
[78] SUN, C.M., DERIAUD, E., LECLERC, C., AND LO-MAN, R. 2005. Upon TLR9 signaling, CD5+ B cells control the IL-12-dependent Th1-priming capacity of neonatal DCs. Immunity. 22:467–477.
[79] SUN S, ZHANG X, TOUGH DF, et al. Type I interferon-mediated stimulation of T cells by CpG DNA. J Exp Med, 1998, 188 (12): 2335- 2342
[80] THOMAS C, FOULET A, PLAT M, et al. E fficacy of rituximab in lymphomatoid granulomatosis. J. Rev Mal Respir, 2004, 21(6): 1157.
[81] THOMAS JP, ARZOOMANIAN RZ, ALBERTI D, et al. PhaseⅠpharmacokinetis and pharmacodynamic atudy of recombinant human endostatin in patients with advanced solid tumors. J. Clin Oncol, 2003, 21 (2): 223-233
[82] TOKUNAGA T, YAMAMOTO H, SHIMADA S, ABE H., FUKUDA T, FUJISAWA Y, FURUTANI Y, YANO O, KATAOKA T, SUDO T, MAKIGUCHI N AND SUGANUMA T. Antitumor activity of deoxyribonucleic acid fraction from Mycobacterium bovis BCG. L. Isolation, physicochemical characterization, and antitimor activity. J. Natl. Cancer Inst., 1984; 72: 955-62
[83] UPPENKAMP M, ENGERT A, DIEHL V, et al. Monoclonal antibody therapy with CAMPATH-1H in patients with relapsed high- and low- grade non-Hodgkin’s lymphomas: a multicenter phase I/ II study. Ann Hematol, 2002, 81: 26-32
[84] UTAISINCHARON P, ANUNTAGOOL N, CHAISURIYA P, PICHYANGKUL S, SIRISINHA S. CpG ODN activates NO and iNOS production in mouse macrophage cell line(RAW 264.7). Clin Exp Immunol. 2002, 128: 467-73
[85] VAN OJIK H, KRUITW, PORTIELJE J, BRICHARD V, VERLOES R, DELIREM. PhaseⅠ/Ⅱstudy with CpG 7909 as adjuvant to vaccination with MAGE-3 protein in patients with MAGE-3 positive tumors. Ann Oncol 13(Suppl). 2002, 157-158
[86] VILLALONA-CALERO MA, RITCH P, FIGUEROA JA, et al. A phaseⅠ/Ⅱstudy of LY900003, an antisense inhibitor of protein kinase C-alpha, in combination with cisplatin and gemcitabine in patients with advanced non-small-cell lung cancer. J. Clin Cancer Res, 2004, 10(18): 6086-6093
[87] VOGEL CL, COBLEIGH MA , TRIPATHY D, et al. First-line herceptin monotherapy in metastatic breast cancer. J. Oncology, 2001, 10(6): 37-42.
[88] VOLLMER J, WEERATNA R, PAYETTE P, JURK M, SCHETTER C, LAUCHT M, WADER T, TLUK S, LIU M, DAVIS HL, KRIEG AM. Characterization of three CpG oligodeoxynucleotide classes with distinct immunostimulatory activities. Eur. J. Immunol. 2004; 34: 251–262
[89] WOOLDRIDGE JE, BALLAS Z, KRIEG AM, WEINER GJ. Immunostimulatory oligodeoxynucleotides containing CpG motifs enhance the efficacy of monoclonal antibody therapy of lymphoma. Blood. 1997; 89(8): 2994-8.
[90] WAAG DM, MCCLUSKIE MJ, ZHANG N, KRIEG AM. A CpG oligonucleotide can protect mice from a low aerosol challenge dose of Burkbolderia mallei. Infect Immun. 2006, 74(3): 1944-8
[91] WAGNER M, POECK H, JAHRSDOERFER B, ROTHENFUSSER S,PRELL D, BOHLE B, TUMA E, GIESE T, ELLWART JW, ENDRES S, HARTMANN G. IL-12p70-dependent Th1 induction by human B cells requires combined activation with CD40 ligand and CpG DNA.J Immunol.2004;172(2):954-63
[92] WAGNER S, et al. CpG 7909, a TLR9 agonist immunomodulator in metastatic melanoma: A randomized phaseⅡtrial comparing two doses and in combination with DTIC [abstract]. In Journal of Clinical Oncology,2005 ASCO Annual Meeting Proceedings. 2006, 23: 7526
[93] WANG, X. S. et al. 2005. CpG oligodeoxynucleotides inhibit tumor growth and reverse the immunosuppression caused by the therapy with 5-fuorouracil in murine hepatoma. World J. Gastroenterol. 11: 1220-1224.
[94] WEIGEL BJ, RODEBERG DA, KRIEG AM. CpG oligodeoxynucleotides potentiate the antitumor effects of chemotherapy or tumor resection in an orthotopic murine model of rhabdomyosarcoma. Clin Cancer Res. 2003. 9(8): 3105-14.
[95] WEINER GJ. The immunobiology and clinical potential of immunostimulatory CpG oligodeoxynucleotides. J Leukoc Biol. 2000; 68(4):455-63
[96] WEINER GJ, LIU HM, WOOLDRIDGE JE, DAHLE CE, KRIEG AM. Lmmunostimulatory oligodeoxynucleotides containing the CpG motif are effective as immune adjuvants in tumor antigen immunization. Proc Natl Acad Sci USA. 1997, 94(20): 10833-7
[97] WEKGEL BJ, PANOSKALTSIS-MORTARI A, DIERS M, GARCIA M, LEES C, KRIEG AM, CHEN W, BLAZAR BR. Dendritic cells pulsed or fused with AML cellular antigen provide comparable in vivo antitumor protective response. Exp Hematol. 2006, 34(10): 1403-12
[98] WILLIAMS KJ, TELFER BA, BRAVE S, et al. ZD6474, a potent inhibitor of vascular endothelial growth factor signaling, combined with radiotherapy: Schedule-dependent enhancement of antitumor activity. J. Clin Cancer Res, 2004, 10(24): 8587-8593
[99] WINGENDER, G., et al. 2006. Systemic application of CpG-rich DNA suppresses adaptive T cell immunity via induction of IDO. Eur. J. Immunol. 36:12–20.
[100] WOOLDRIDGE JE, WEINER GJ. CpG DNA and cancer immunotherapy. Orchestrating the antitumor immune response. Curr Opin Oncol.2003. 15(6):440-5.
[101] YAMAMOTO S,KURAMOTO E,SHIMADA S,TOKUNAGA T.In vitro augmentation of natural killer cell activity and production of interferon-alpha/beta and-gamma with deoxyribonucleic acid fraction from Mycobacterium bovis BCG.Jpn J Cancer Res.1988;79(7):866-73
[102] YAMAMOTO M, TAKEDA K, AKIRA S. TIR domain-containing adaptors define the specificity of TLR signaling. J Mol Immunol, 2004, 40: 861-868
[103] YI AK, PECKHAM DW, A SHMAN RF, et al. CpG DNA rescues B Cells from apoptosis by activating NF kappa B and preventing mitochondrial membrane potential disruption via a chloroquine-sentive pathway [J ]. Int Immunol, 1999, 11 (12): 2015- 2024.
[104] YI AK, YOON JG, KRIEG AM. Convergence of CpG DNA-and BCR-mediated signals at the c-Jun N-terminal kinase and NF-kappaB activation pathways: regulation by mitogen-activated protein kinases. Int Immunol.2003; 15(5): 577-91
[105]史鹤玲,徐丽艳,刘哲.重组人血管内皮抑制素(YH-16)注射液治疗晚期非小细胞肺癌Ⅱ期临床研究[J ] .中国肺癌杂志,2004 ,7 (4)∶325– 328
[106]张青松,刘林强,王玲.肺癌疫苗的研究进展.医学伦理与实践,2004,17(10):1140～1141
[2] AKIRA S, TAKEDA K. Toll-like receptor signalling. Nat Rev Immunol. 2004; 4(7): 499-511
[3] BABATZ J, ROLLIG C, LOBEL B, et al. Induction of cellular immune responses against carcinoembryonic antigen in patients with metastatic tumors after vaccination with altered peptide ligand-loaded dendritic cells. J. Cancer Immunol Immunother, 2005, 54(2): 1-9
[4] BALLAS ZK, RASMUSSEN WL, KRIEG AM. Induction of NK activity in murine and HUMAN cells by CpG motifs in oligodeoxynucleotides and bacterial DNA. J Immunol. 1996, 157(5): 1840-5.
[5] BALSARI A, TORTORETO M, BESUSSO D, PETRANGOLINI G, SFONDRINI L, MAGGI R, MENARD S, PRATESI G. Combination of a CpG-oligodeoxynucleotide and a topoisomerase I inhibitor in the therapy of human tumour xenografts. Eur J Cancer. 2004 May; 40(8): 1275-81.
[6] BAUER S, KIRSCHNINQ CJ , HACKER H. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. Proc Nal A cad Sci U S A , 2001, 98 (16): 9237-9242
[7] BENDIGS S, SALZER U, LIPFORD GB, et al. CpG-oligodeoxynucleotides co-stimulate primary T cells in the absence of antigen-presenting cells. Eur J Immunol, 1999, 29 (4): 1209-1218.
[8] BERNASCONI N.L., ONAI N. & LANZAVECCHIA A. A role for Toll-like receptors in acquired immunity:up-regulation of TLR9 by BCR triggering in naive cells and constitutive expression in memory B cells Blood ,2003,101, 4500–4504.
[9] BISSETT D, O′BYRNE KJ, VON PAWEL J, et al. PhaseⅢstudy of matrix metalloproteinase inhibitor prinomastat in non-small-cell lung cancer. J. Clin Oncol, 2005, 23 (4): 842-849.
[10] BRENNAN FR, BELLABY T, HELLIWELL SM, et al. Chimeric plant virus particles administered nasally or orally induce systemic and mucosal immune responses in mice. J Virol, 1999, 73(2): 930-938.
[11] BRUNNER C, SEIDERER J, SCHLAMP A, BIDLINGLNAIER M, EIGLER A, HAIMERL W, et al. Enhanced dendritic cell maturation by TNF-alpha or cytidine-phosphate-guanosine DNA drives T cell activation in vitro and therapeutic anti-tumor immune responses in vivo. J Immunol. 2000, 165: 6278-86.
[12] CARPENTIER A, LAIGLE-DONADEY F, ZOHAR S, CAPELLE L, BEHIN A, TIBI A, MARTIN-DUVERNEUIL N, SANSON M, LACOMBLEZ L, TAILLIBERT S, PUYBASSET L, VAN EFFENTERRE R, DELATTRE JY, CARPENTIER AF. Phase 1 trial of a CpG oligodeoxynucleotide for patients with recurrent glioblastoma. Neuro Oncol. 2006, 8(1): 60-6.
[13] CHACE JH, HOOKER NA, MILDENSTEIN KL, KRIEG AM, COWDERY JS. Bacterial DNA-induced NK cell IFN-gamma production is dependent on macrophage secretion of IL-12. Clin immunol Immunopathol. 1997, 84: 185-93.
[14] COLEY WB. The treatment of malignant tumors by repeated inoculations of erysipelas with a report of ten original cases. Am J Med Sci. 1893; 105: 487–511.
[15] DAVILA E, KENNEDY R, CELIS E. Generation of antitumor immunity by cytotoxic T lymphocyte epitope peptide vaccination, CpG-oligodeoxynucleotide adjuvant and CTLA-4 blockade. Cancer Res. 2003. 63(12): 3281-8.
[16] DECKER T, HIPP S, KREITMAN RJ, PASTAN I, PESCHEL C, LICHT T. Sensitization of B-cell chronic lymphocytic leukemia cells to recombinant immunotoxin by immunostimulatory phosphorothioate oligodeoxynucleotides. Blood. 2002. 99 (4): 1320-6.
[17] DROEMANN D, ALBRECHT D, GERDES J, ULMER AJ, BRANSCHEID D, VOLLMER E, DALHOFF K, ZABEL P, GOLDMANN T. Human lungcancer cells express functionally active Toll-like receptor 9. Respir Res. 2005 Jan 4; 6:1.
[18] DUNNE A, O’NEILL LAJ. The interleukin-1 receptor/toll-like receptor superfamily: signal transduction during inflammation and host defense. J Science’s STKE, February 25, 2003, 2003(171): re3-3
[19] EL ANDALOUSSI A, SONABEND AM, HAN Y, LESNIAK MS. Stimulation of TLR9 with CpG ODN enhances apoptosis of glioma and prolongs the survival of mice with experimental brain tumors. Glia. 2006 Nov 1; 54(6): 526-35.
[20] FADERL S, THOMAS DA, O′BRIEN S, et al. Experience with alemtuzumab plus rituximab in patients with relapsed and refractory lymphoid malignancies. Blood, 2003, 101: 3413-3415.
[21] FELDMAN E, KALAYCIO M, WEINER G, et al. Treatment of relapsed or refractory acute myeloid leukemia with humanized anti-CD33 monoclonal antibody HuM195. Leukemia, 2003, 17: 3142 318
[22] FRIEDBERG JW, KIM H, MCCAULEY M, HESSEL EM, SIMS P, FISHER DC, NADLER LM, COFFMAN RL, FREEDMAN AS. Combination immunotherapy with a CpG olgonucleotide (1018 ISS) and rituximab in patients with non-Hodgkin lymphoma: increased interferon-alpha/beta-inducible gene expression, without significant toxicity. Blood. 2005, 105(2): 489-95
[23] FUKUOKA M, YANO S, GIACCONE G, et al. Multi-institutional randomized phaseⅡtrial of gefinitib for previously treated patients with advanced non-small-cell lung cancer. J Clin Oncol, 2003, 21 (12): 2237.
[24] GATZEMEIER U, GROTH G, BRITTS C, et al. Randomized phaseⅡtrial of Gemcitabine -Cisplatin with or without trastuzumab in Her-2 positive non-small cell lung cancer. J. Ann Oncol, 2004, 15(1):19-27.
[25] GRIDELLI C, MAIONE P, DEL GAIZO F, et al. Sorafenib and sunitinib in the treatment of advanced non-small-cell lung cancer. Oncologist, 2007, 12(2): 191-200
[26] GROSSBARD ML, MULTANI PS, FREEDMAN AS, et al. A Phase II study of adjuvant therapy with anti-B4-blocked ricin after autologous bone marrow transplantation for patients with relapsed B-cell non-Hodgkin′s lymphoma. Clin Cancer Res, 1999, 5:2392-2398
[27] HALE G, SLAVIN S, GOLDMAN JM. Alemtuzumab (Campath-1H) for treatment of lymphoid malignancies in the age of nonmyeloablative conditioning? Bone Marrow Transplant, 2002, 30: 797-804.
[28] HARTMANN G, WEERATNA RD, BALLAS ZK, PAYETTE P, BLACKWELL S, SUPARTO I, RASMUSSEN WL, WALDSCHMIDT M, SAJUTHI D, PURCELL RH, DAVIS HL, KRIEG AM. Delineation of a CpG Phosphorothioate Oligodeoxynucleotide for Activating Primate Immune Responses In Vitro and In Vivo. J Immunol. 2000; 164: 1617–1624
[29] HE H, CRIPPEN TL, FARNELL MB, KOGUT MH. Identification of CpG oligodeoxynucleotide motifs that stimulate nitric oxide and cytokine production in avian macrophage and peripheral blood mononuclear cells. Dev Comp Immunol. 2003, 27: 621-7
[30] HERBST RS, GIACCONE G, SCHILLER JH, et al. Gefitinib in combination with paclitaxel and carboplatin in advanced non-small-cell lung cancer: a phaseⅢtrial- INTACT 2 . J Clin Oncol, 2004, 22 (5): 785.
[31] HERBST RS, PRAGERD, HERMAN R, et al. TRIBUTE: A phaseⅢtrial of erlotinib hydrochloride (OSI-774) combined with carboplatin and paclitaxel chemotherapy in advanced non-small–cell lung cancer. J Clin Oncol, 2005, 23 (25): 5892.
[32] HEYMACH J, WEST H, KERR R, et al. ZD6474 in combination with carbopatin and paclitaxel as first line treatment in patients with NSCLC: Results of the run- in phase of a two-part randomized phaseⅡstudy. J. Lung Cancer, 2005, 49(Supp 12): S247.
[33] HURWITZ H, HOLDEN SN, ECKHARDT SG, et al. Clinical evaluation of ZD6474, an orally active inhibitor of VEGF signaling, in patients with solid tumors. J. Proc Am Soc Clin Oncol, 2002, 21: 82a
[34] IHO S, YAMAMOTO T, TAKAHASHI T, YAMAMOTO S. Oligodeoxynucleotides containing palindrome sequences with internal 5’-CpG-3’act directly on human NK and activated T cells to induce IFN-gamma production in vitro. J Immunol. 1999, 163: 3642-52
[35] JAHRSDORFER B, JOX R, MUHLENHOFF L, TSCHOEP K, KRUG A, ROTHENFUSSER S, MEINHARDT G, EMMERICH B, ENDRES S, HARTMANN G. Modulation of malignant B cell activation and apoptosis by bcl-2 antisense ODN and immunostimulatory CpG ODN. J Leukoc Biol. 2002, 70: 83-92
[36] JAHRSDORFER B, MüHLENHOFF L, BLACKWELL SE, WAGNER M, POECK H, HARTMANN E, JOX R, GIESE T, EMMERICH B, ENDRES S, WEINER GJ, HARTMANN G. B-cell lymphomas differ in their responsiveness to CpG oligodeoxynucleotides. J. Clin Cancer Res. 2005 Feb 15; 11(4): 1490-9.
[37] JAHRSDORFER, B. et al. 2005. Immunostimulatory oligodeoxynucleotides induce apoptosis of B cell chronic lymphocytic leukemia cells. J. Leukoc. Biol. 77: 378-387.
[38] JOHNSON DH, FEHRNBACHER L, NOVOTNY WF, et al. Randomized phaseⅡtrial comparing bevacizumb plus carboplatin and paclitaxel with carboplatin and paclitaxel alone in previously untreated locally advanced or metastic non-small-cell lung cancer. J Clin Oncol, 2004 ,22 (11): 2184-2191.
[39] KADOWAKI N, ANTONENKO S, LIU YJ. Distinct CpG DNA and polyinosinic-polycytidylic acid double-stranded RNA, respectively, stimulate CD11c-type 2 dendritic cell precursors and CD11c+ dendritic cells to produce type I IFN. J Immunol.2001 Feb 15; 166(4): 2291-5.
[40] KAWARADA Y, GANSS R, GARBI N, SACHER T, AMOLD B, H?MMERLING GJ. NK+ and CD8+ T Cell-Mediated Eradication of Established Tumors by Peritumoral Injection of CpG-Containing Oligodeoxynucleotides J Innunol. 2001, 167(9): 5247-53
[41] KIM, Y, GIRARDI M, DUVIC M, KUZEL T, ROOK A, LINK B. TLR9agonist immunomodulator treatment of cutaneous T-cell lymphoma(CTCL) with CpG 7909. ASH Annual Meeting Abstracts 104. 2004, Abstract 743
[42] KLINMAN DM,YI AK, BEAUCAGE SL, CONOVER J,KREIG AM. CPG motifs Present in bacteria DNA rapidly induce lymphocytes to secrete interleukin 6,interleukin 12, and interferon gamma. Proc Natl Acad Sci USA. 1996, 93: 2879-83
[43] KOHLER G, MILSTEIN C. Continuous cultures of fused cells secreting antibody of predefined specificity. J Immunol, 2005, 174(5): 2453.
[44] KRIEG AM. Toll-like receptor9 (TLR9) agonists in the treatment of cancer. Oncogene. 2008, 27(2): 161-7
[45] KRIEG AM. Therapeutic potential of Toll-like receptor 9 activation. Nat Rev Drug Discov. 2006 Jun; 5(6): 471-84.
[46] KRIEG AM,YI AK,MATSON S,WALDSCHMIDT TJ,BISHOP GA,TEASDALE R,KORETZKY GA,KLINMAN DM.CpG motifs in bacterial DNA trigger direct B-cell activation.Nature.1995;374:546-549
[47] KRIS MG, NATALE RB, HERBST RS, et al. A phaseⅡtrial of ZD1839 (Iressa) in advanced non-small-cell lung cancer (NSCLC) patients who had failed platinum-and docetaxel-based regi mens (IDEAL-2). J. Pro Am Soc Clin Oncol, 2002, 21: 292.
[48] KOLUMAM GA, S THOMAS, L J THOMPSON, J SPRENT, K MURALI-KRISHNA. TypeⅠinterferons act directly on CD8 T cells to allow clonal expansion and memory formation in response to viral infection. J Exp Med. 2005, 202: 637-650
[49] LANGER C J, STEPHENSON P, THOR A, et al. Trastuzumab in the treatment of advanced non-small-cell lung cancer : Is there a role? Focus on Eastern Cooperative Oncology Group Study 2598. J. J Clin Oncol, 2004, 22 (7): 1180-1187.
[50] LATZ E, SCHOENEMEYER A, VISINTINA, FITZGERALD KA, MONKS BG, KNETTER CF, LIEN E, NILSEN NJ, ESPEVIK T, GOLENBOCK DT. TLR9 signals after translocating from the ER to CpG DNA in the lysosome.Nat lmmunol. 2004, 5(2): 190-8.
[51] LEIGHL NB, PAZ-ARES L, DOUILLARD JY, et al. Randomized phaseⅢstudy of matrix metalloproteinase inhibit or BMS-275291 in combination with paclitaxel and carboplatin in advanced non-small-cell lung cancer: National Cancer Institute of Canada-Clinical Trials Group Study BR. 18. J. Clin Oncol, 2005, 23 (12): 2831-2839
[52] LINK BK, BALLAS ZK. WEISDORF D, WOOLDRIDGE JE, BOSSLER AD, SHANNON M, RASMUSSEN WL, KRIEG AM, WEINER GJ. Oligodeoxynucleotide CPG7909 delivered as intravenous infusion demonstrates immunologic modulation in patients with previously treated non-Hodgkin lymphoma. J. Immunother. 2006, 29(5): 558-68
[53] MANEGOLD C, et al. Addition of PF-3512676 (CpG 7909) to a taxane/platinum regimen for first-line treatment of unresectable non-small cell lung cancer (NSCLC) improves objective response-phaseⅡclinical trial [abstract]. Eur J Cancer. 2005, 3: 326
[54] MARRACK, P., J. Kappler, and T. Mitchell. TypeⅠinterferons keep activated T cells alive. J. Exp. Med. 1999, 189: 521-530
[55] MASON KA, ARIGA H, NEAL R, et al. Targeting toll-like receptor 9 with CpG oligodeoxynucleotides enhances tumor response to fractionated radiotherapy. J. Clin Cancer Res, 2005, 11(1): 361-369
[56] MASON KA, NEAL R, HUNTER N, et al. CpG oligodeoxynucleotides are potent enhancers of radio- and chemo responses of murine tumors. J. Radio ther Oncol, 2006, 80 (2): 192-198
[57] MELLOR, A.L., et al. 2005. Cutting edge: CpG oligonucleotides induce splenic CD19+ dendritic cells to acquire potent indoleamine 2, 3-dioxygenase-dependent T cell regulatory functions via IFN Type 1 signaling. J. Immunol. 175:5601–5605.
[58] MILAS L, MASON KA, ARIGA H, HUNTER N, NEAL R, VALDECANAS D, KRIEG AM, WHISNANT JK. CpG oligodeoxynucleotide enhances tumor response to radiation. Cancer Res. 2004, 64(15): 5074-7
[59] MODELSKA A, DIETZSCHOLD B, SLEYSH N, et al. Immunization against rabies with plant-derived antigen. J Proc Natl Acad Sci USA, 1998, 95(5): 2481-2485.
[60] MOLENKAMP BG, VAN LEEUWEN PA, VAN DEN EERTWEGH AJ, SLUIJTER BJ, SCHEPER RJ, MEIJER S, DE GRUIJL TD. Immunomoeulation of the melanoma sentinel lymph node: a novel adjuvant therapeutic option. Immunobiology. 2006, 211(6-8): 651-61
[61] NATALE R, BODKIN D, GOVINDAN R, et al. A comparison of the antitumor efficacy of ZD6474 and gefitinib (Iressa) in patients with NSCLC: results of a randomized, double blind, phaseⅡstudy. Lung Cancer, 2005, 49 (2): s371
[62] NIETHAMMER AG, PRIMUS FJ, XIANG R, et al. An oral DNA vaccine against human carcinoembryonic antigen (CEA) prevents growth and dissemination of Lewis lung carcinoma in CEA transgenic mice. Vaccine, 2001,20(3-4): 421-429
[63] ONN A, CORREA AM, GILCREASE M, et al. Synchronous over-expression of epidermal growth factor receptor and HER2-neu protein is a predict or of poor outcome in patients with stage I non-small cell lung cancer. Clin Cancer Res, 2004, 10 (1) : 136-143
[64] ONO M, KUWANO M. Molecular mechanisms of epidermal growth factor receptor (EGFR) activation and response to gefitinib and other EGFR-targeting drugs. J. Clin Cancer Res, 2006, 12 (24): 7242-7251.
[65] PASHENKOV M, GOЁSSG, WAGNER C, H?RMANN M, JANDL T, MOSER A, BRITTEN CM, SMOLLE J, KOLLER S, MAUCH C, TANTCHEVA-POOR L, GRABBE S, LOQUAI C, ESSER S, FRANCKSON T, SCHNEEBERGER A, HAARMANN C, KRIEG AM, STINGL G, WAGNER SN. PhaseⅡtrial of a toll-like receptor 9-activating oligonueleotide in patients with metastatic melanoma. J. Clin Oncol. 2006, 24(36): 5716-24
[66] PICCALUGA P P, ARTINELL I G, RONDON IM, et al. Gemtuzumabozogamicin for relapsed and refractory acute myeloid leukemia and myeloid sarcomas. J Leuk Lymphoma, 2004, 45 (9): 1791-1795
[67] PILON-THOMAS S, LI W, BRIGGS JJ, DJEU J, MULéJJ, RIKER AI. lmmunostimulatory effects of CpG-ODN upon dendritic cell-based immunotherapy in a murine melanoma model. J Immunother (1997). 2006, 29(4): 381-7
[68] RUTZ M, METZGER J, GELLERT T, LUPPA P, LIPFORD GB, WAGNER H, BAUER S. Toll-like receptor 9 binds single-stranded CpG-DNA in a sequence-and pH-dependent manner. Eur J Immunol. 2004 Sep; 34(9): 2541-50.
[69] ROSELL R, GATZEMEIER U, BETTICHER DC, et al. PhaseⅢrandomized trial comparing paclitaxel/carboplatin with paclitaxel/cisplatin in patients with advanced non-small-cell Lung cancer: A cooperative multinational trial. J. Ann Oncol, 2002, 13(10): 1539-1549
[70] SCHATTENBERG D, SCHOTT M, REINDL G, KRUEGER T, TSCHOEPE D, FELDKAMP J, SCHERBAUM WA, SEISSLER J. Response of human monocyte-derived dendritic cells to immunostimulatory DNA. Eur J Immunol. 2000; 30(10): 2824-31.
[71] SCHILLER JH, HARRINGTON D, BELANI CP, et al. Comparison of four chemotherapy regimens for advanced non- small- cell lung cancer. J. N Engl J Med, 2002, 346(2): 92-98
[72] SHARMA S, KARAKOUSIS CP, TAKITA H, SHIN K, BROOKS SP. Intra-tumoral injection of CpG results in the inhibition of tumor growth in murine Colon-26 and B-16 tumors. Biotechnol Lett. 2003, 25(2): 149-53
[73] SHEPHERD FA, RODRIGUES PEREIRA J, CIULEANU T, et a1. Erlotinib in previously treated non-small-cell lung cancer. N Engl J Med, 2005, 353 (2): 123-132
[74] SHEPHERD FA, PEREIRA J, CIULEANU JE, et al. A randomized placebo-controlled trial of erlotinib in patients with advanced non-small-cell lung cancer (NSCLC) following failure of 1st line or 2nd line chemotherapy. ANational Cancer Institute of Canada Clinical Trials Group (NCICCTG) Trial. J. Proc Am Soc Clin Oncol, 2004, 23: Abs 7022
[75] SHODA LK, KEGERREIS KA, SUAREZ CE, MWANGI W, KNOWLES DP, BROWN WC. Immunostimulatory CpG-modified plasmid DNA enhances IL-12, TNF-alpha, and NO production by bovine macrophages. J Leukoc Biol. 2001, 70(l): 103-12
[76] SIEVERS E L, LARSON R A, STADTMAUER E A, et al. Efficacy and safety of gemtuzumab ozogamicin in patients with CD33-positive acute myeloid leukemia in first relapse. J Clin Oncol, 2001, 19 (13): 3244-3254
[77] SPEISER DE, LIéNARD D, RUFER N, RUBIO-GODOY V, RIMOLDI D, LEJEUNE F, KRIEG AM, CEROTTINI JC, ROMERO P. Rapid and strong human CD8+ T cell responses to wanccination with peptide, IFA, and CpG oligodeoxynucleotide 7909. J Clin Invest. 2005, 115(3):739-46
[78] SUN, C.M., DERIAUD, E., LECLERC, C., AND LO-MAN, R. 2005. Upon TLR9 signaling, CD5+ B cells control the IL-12-dependent Th1-priming capacity of neonatal DCs. Immunity. 22:467–477.
[79] SUN S, ZHANG X, TOUGH DF, et al. Type I interferon-mediated stimulation of T cells by CpG DNA. J Exp Med, 1998, 188 (12): 2335- 2342
[80] THOMAS C, FOULET A, PLAT M, et al. E fficacy of rituximab in lymphomatoid granulomatosis. J. Rev Mal Respir, 2004, 21(6): 1157.
[81] THOMAS JP, ARZOOMANIAN RZ, ALBERTI D, et al. PhaseⅠpharmacokinetis and pharmacodynamic atudy of recombinant human endostatin in patients with advanced solid tumors. J. Clin Oncol, 2003, 21 (2): 223-233
[82] TOKUNAGA T, YAMAMOTO H, SHIMADA S, ABE H., FUKUDA T, FUJISAWA Y, FURUTANI Y, YANO O, KATAOKA T, SUDO T, MAKIGUCHI N AND SUGANUMA T. Antitumor activity of deoxyribonucleic acid fraction from Mycobacterium bovis BCG. L. Isolation, physicochemical characterization, and antitimor activity. J. Natl. Cancer Inst., 1984; 72: 955-62
[83] UPPENKAMP M, ENGERT A, DIEHL V, et al. Monoclonal antibody therapy with CAMPATH-1H in patients with relapsed high- and low- grade non-Hodgkin’s lymphomas: a multicenter phase I/ II study. Ann Hematol, 2002, 81: 26-32
[84] UTAISINCHARON P, ANUNTAGOOL N, CHAISURIYA P, PICHYANGKUL S, SIRISINHA S. CpG ODN activates NO and iNOS production in mouse macrophage cell line(RAW 264.7). Clin Exp Immunol. 2002, 128: 467-73
[85] VAN OJIK H, KRUITW, PORTIELJE J, BRICHARD V, VERLOES R, DELIREM. PhaseⅠ/Ⅱstudy with CpG 7909 as adjuvant to vaccination with MAGE-3 protein in patients with MAGE-3 positive tumors. Ann Oncol 13(Suppl). 2002, 157-158
[86] VILLALONA-CALERO MA, RITCH P, FIGUEROA JA, et al. A phaseⅠ/Ⅱstudy of LY900003, an antisense inhibitor of protein kinase C-alpha, in combination with cisplatin and gemcitabine in patients with advanced non-small-cell lung cancer. J. Clin Cancer Res, 2004, 10(18): 6086-6093
[87] VOGEL CL, COBLEIGH MA , TRIPATHY D, et al. First-line herceptin monotherapy in metastatic breast cancer. J. Oncology, 2001, 10(6): 37-42.
[88] VOLLMER J, WEERATNA R, PAYETTE P, JURK M, SCHETTER C, LAUCHT M, WADER T, TLUK S, LIU M, DAVIS HL, KRIEG AM. Characterization of three CpG oligodeoxynucleotide classes with distinct immunostimulatory activities. Eur. J. Immunol. 2004; 34: 251–262
[89] WOOLDRIDGE JE, BALLAS Z, KRIEG AM, WEINER GJ. Immunostimulatory oligodeoxynucleotides containing CpG motifs enhance the efficacy of monoclonal antibody therapy of lymphoma. Blood. 1997; 89(8): 2994-8.
[90] WAAG DM, MCCLUSKIE MJ, ZHANG N, KRIEG AM. A CpG oligonucleotide can protect mice from a low aerosol challenge dose of Burkbolderia mallei. Infect Immun. 2006, 74(3): 1944-8
[91] WAGNER M, POECK H, JAHRSDOERFER B, ROTHENFUSSER S,PRELL D, BOHLE B, TUMA E, GIESE T, ELLWART JW, ENDRES S, HARTMANN G. IL-12p70-dependent Th1 induction by human B cells requires combined activation with CD40 ligand and CpG DNA.J Immunol.2004;172(2):954-63
[92] WAGNER S, et al. CpG 7909, a TLR9 agonist immunomodulator in metastatic melanoma: A randomized phaseⅡtrial comparing two doses and in combination with DTIC [abstract]. In Journal of Clinical Oncology,2005 ASCO Annual Meeting Proceedings. 2006, 23: 7526
[93] WANG, X. S. et al. 2005. CpG oligodeoxynucleotides inhibit tumor growth and reverse the immunosuppression caused by the therapy with 5-fuorouracil in murine hepatoma. World J. Gastroenterol. 11: 1220-1224.
[94] WEIGEL BJ, RODEBERG DA, KRIEG AM. CpG oligodeoxynucleotides potentiate the antitumor effects of chemotherapy or tumor resection in an orthotopic murine model of rhabdomyosarcoma. Clin Cancer Res. 2003. 9(8): 3105-14.
[95] WEINER GJ. The immunobiology and clinical potential of immunostimulatory CpG oligodeoxynucleotides. J Leukoc Biol. 2000; 68(4):455-63
[96] WEINER GJ, LIU HM, WOOLDRIDGE JE, DAHLE CE, KRIEG AM. Lmmunostimulatory oligodeoxynucleotides containing the CpG motif are effective as immune adjuvants in tumor antigen immunization. Proc Natl Acad Sci USA. 1997, 94(20): 10833-7
[97] WEKGEL BJ, PANOSKALTSIS-MORTARI A, DIERS M, GARCIA M, LEES C, KRIEG AM, CHEN W, BLAZAR BR. Dendritic cells pulsed or fused with AML cellular antigen provide comparable in vivo antitumor protective response. Exp Hematol. 2006, 34(10): 1403-12
[98] WILLIAMS KJ, TELFER BA, BRAVE S, et al. ZD6474, a potent inhibitor of vascular endothelial growth factor signaling, combined with radiotherapy: Schedule-dependent enhancement of antitumor activity. J. Clin Cancer Res, 2004, 10(24): 8587-8593
[99] WINGENDER, G., et al. 2006. Systemic application of CpG-rich DNA suppresses adaptive T cell immunity via induction of IDO. Eur. J. Immunol. 36:12–20.
[100] WOOLDRIDGE JE, WEINER GJ. CpG DNA and cancer immunotherapy. Orchestrating the antitumor immune response. Curr Opin Oncol.2003. 15(6):440-5.
[101] YAMAMOTO S,KURAMOTO E,SHIMADA S,TOKUNAGA T.In vitro augmentation of natural killer cell activity and production of interferon-alpha/beta and-gamma with deoxyribonucleic acid fraction from Mycobacterium bovis BCG.Jpn J Cancer Res.1988;79(7):866-73
[102] YAMAMOTO M, TAKEDA K, AKIRA S. TIR domain-containing adaptors define the specificity of TLR signaling. J Mol Immunol, 2004, 40: 861-868
[103] YI AK, PECKHAM DW, A SHMAN RF, et al. CpG DNA rescues B Cells from apoptosis by activating NF kappa B and preventing mitochondrial membrane potential disruption via a chloroquine-sentive pathway [J ]. Int Immunol, 1999, 11 (12): 2015- 2024.
[104] YI AK, YOON JG, KRIEG AM. Convergence of CpG DNA-and BCR-mediated signals at the c-Jun N-terminal kinase and NF-kappaB activation pathways: regulation by mitogen-activated protein kinases. Int Immunol.2003; 15(5): 577-91
[105]史鹤玲,徐丽艳,刘哲.重组人血管内皮抑制素(YH-16)注射液治疗晚期非小细胞肺癌Ⅱ期临床研究[J ] .中国肺癌杂志,2004 ,7 (4)∶325– 328
[106]张青松,刘林强,王玲.肺癌疫苗的研究进展.医学伦理与实践,2004,17(10):1140～1141