CpG ODN活化外周血单个核细胞对淋巴瘤细胞作用的研究
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摘要
背景和目的
恶性淋巴瘤(Malignant Lymphoma, ML)是原发于淋巴结或淋巴结外组织或器官的一种恶性肿瘤,来源于淋巴细胞及其前体细胞。在我国恶性淋巴瘤虽相对少见,但近年来新发病例逐年上升,每年至少超过25000例,已成为严重威胁人类健康的一类疾病。恶性淋巴瘤的治疗以化疗为主,同时配合放疗和生物治疗等综合治疗。尽管一部分恶性淋巴瘤经过化疗等综合治疗后可以达到长期持续缓解,但总的来说恶性淋巴瘤因为其侵袭性、原发和继发耐药等原因,治疗效果往往不尽人意。因此,恶性淋巴瘤的攻克需要发展新的治疗途径与手段。目前研究较多的是肿瘤的免疫治疗,其中CpG ODN是肿瘤免疫治疗研究的热点。现已有多篇文献报道,CpG ODN对多种实体瘤的免疫治疗效果显著,但有关淋巴造血系统的恶性肿瘤研究较少。
过去几十年里大量的证据显示,病原体中的核酸不仅能作为生命的基本物质而存在,而且还可以激活宿主的免疫系统。细菌中未甲基化5'CpG3'二核苷酸(CpGoligodeoxynucleotide, CpG ODN)对各种免疫细胞亚群有强大的免疫激活作用。它们能直接激活巨噬细胞、单核细胞和树突状细胞等抗原递呈细胞,释放多种细胞因子,如白介素(IL)-12和IL-6等,再由这些细胞因子间接激活自然杀伤(NK)细胞及T细胞,发挥抗病毒、抗细菌感染及抗肿瘤作用。
本研究运用人工合成的含有CpG基序的寡聚脱氧核苷酸刺激人外周血单个核细胞(Peripheral blood mononuclear cell,PBMC),观察其对PBMC的活化刺激作用,以及由此介导的对淋巴瘤hut-78细胞的杀伤作用,旨在为淋巴瘤的免疫治疗提供一种新途径。
方法
淋巴细胞分离液分离健康人外周抗凝血,体外细胞培养技术常规培养PBMC；将实验分为四组：①阴性对照组：PBS+PBMC;②阳性对照组：LPS+PBMC；③实验组：CpG ODN+PBMC；④联合组：CpG ODN+LPS+PBMC.ELISA法检测CpG ODN活化PBMC后分泌IL-12.INF-γ的含量,流式细胞仪测活化PBMC表面标记CD80、CD86分子的表达；MTT法检测CpG ODN活化PBMC对hut-78细胞杀伤作用,倒置显微镜下观察细胞形态学变化。
结果
1 CpG ODN活化后PBMC培养液上清中IL-12和INF-γ的表达水平
CpG ODN活化后PBMC培养液上清中IL-12和INF-γ的表达水平显著升高。经LPS组、CpG ODN组及LPS+ CpG ODN组活化后的PBMC产生IL-12的含量分别60.54±1.82 pg.mL-1、115.50±19.34 pg.mL-1、120.75±13.71 pg.mL-1；产生INF-γ的含量分别为158.74±18.46 pg.mL-1、262.75±30.21 pg.mL-1、265.25±29.26 pg.mL-1。实验组结果与空白对照组差异有统计学意义(P<0.01)。
2 CpG ODN诱导PBMC表面标志的表达
与对照组相比,PBMC在经LPS组、CpG ODN组刺激后细胞表面标志协同刺激分子CD80、CD86的表达水平均明显增高(P<0.01)；CpG ODN组刺激的PBMC表达协同刺激分子CD80、CD86明显大于阳性对照LPS组(P<0.05)。
3 CpG ODN活化的PBMC对HUT-78细胞杀伤作用
CpG ODN活化后的PBMC对HUT-78细胞杀伤活力显著提高。经LPS组.CpG ODN组及CpG ODN+ LPS组活化后的人外周血单个核细胞对HUT-78细胞杀伤活力分别为41.86±2.93%、63.65±7.98%、67.05±3.56%。其实验组0D值与空白对照组差异有统计学意义(P<0.05)。
4形态学观察结果
通过观察发现HUT-78细胞与经CpG ODN刺激后的PBMC共同培养48小时后,形态学改变较明显：有些细胞边界不清,相互融合,部分细胞可见核浓缩。
结论
1经CpG ODN2006活化PBMC分泌细胞因子IL-12和INF-γ水平显著提高。
2 CpG ODN刺激PBMC表面表达的协同刺激分子CD80、CD86水平明显增高
3 CpG ODN显著增强人外周血单个核细胞对HUT-78细胞杀伤活力。
Background and Objective
Malignant lymphoma is a malignant proliferative disease generates from the human immune system cells and their precursor cells. Although malignant lymphoma in China is relatively rare, yet new cases have been increasing in recent years, at least more than 25,000 cases a year, which often occurs in young and seriously endangers the health and lives of human beings. Currently, combined chemotherapy is the common modality of the treatment of malignant lymphoma. It plays an important role in improving the curative rate and remarkably prolonging the disease-free interval of sufferers, but it has inevitably adverse reactions, such as bone marrow suppression and the decreasing of immune system, which make it difficult for the patients to complete the full course of treatment. Moreover, Primary or secondary drug resistance during treatment has become one of the tough problems of clinical therapy of the malignant tumor. Therefore, it is time to seek new ways and means of treatment for malignant lymphoma. More current researches are focus on immunotherapy of cancer and CpG ODN. A number of articles have been published that treatment with CpG ODN on solid tumors was significantly. However, the study of malignant tumors of lymphoid system is less.
DNAs in bacteria and virus, which contain numethylated CPG motifs activate host defense mechanisms leading to innate and acquired immune responses. Synthetic oligodeoxynucleotides(ODNs) containing unmethylated CPG motifs Minic the immunostimulatory qualities of bacterial DNA and elicit a coordinated set of T helper 1 (Thl)-like immune responses, including direct activation of B cells, plasmacytoid dendritic cells (PDCs), macrophages, antigen present cell (APC) and indirect activation of T cells, natural Killer (NK) cells. CpG ODN-induced activation of innate immunity protects against lethal challenge with a wide variety of pathogens. However, we can only find some reports on antitumor activity of CPG ODN in solid tumors.
So, in this study, we will investigate the antitumor effects of PBMC activated by Synthetic oligodeoxynucleotides (ODN) containing unmethylated CPG dinucleotides (CPG ODN) in vitro and explore the mechanisms of anti-tumor action of CPG ODN, which offers a new way for the treatment of the lymphoma.
Methods
Separating the circumference anti-hemoglutination of healthy persons with Lymphocyte separation medium, human peripheral blood mononuclear cells (PBMC) were cultured with technology of cell culture in vitro. We established four groups in the experiment:control group:PBMC+PBS; Positive control group:PBMC+LPS; ODN2006 group:PBMC+ODN2006; Combination group:PBMC+ODN2006+LPS. The secretions of INF-γand IL-12 from PBMC determined using ELISA method. After treated with these groups for 48 hours, Co-stimulatory molecules of CD80, CD86 on the surface of PBMC were analyzed by flow cytometry. PBMC activated by CPG ODN was co-cultured with hut-78 cells for 48 hours, and then MTT method was used to detect PBMC-mediated antitumor activity. We examined cell morphology with the inverted microscope.
Results
1. The effect of CpG ODN on INF-γand IL-12 of PBMC
The secretions of INF-γand IL-12 from PBMC treated with CpG ODN were increased significantly. The levers of IL-12 from PBMC of control group, Positive control group, CpG ODN group and Combination group was 15.50±10.12, 60.54±1.82 pg-mL-1,115.50±19.34 pg·mL-1,120.75±13.71 pg·mL-1,respectively; The levers of INF-γfrom PBMC of these four groups was 85.50±10.01, 158.74±18.46 pg·ml-1,262.75±30.21 pg·mL-1,265.25±29.26 pg·mL-1 separately.
2. The effect of PBMC treated with CpG ODN on Co-stimulatory molecules of CD80, CD86 of PBMC
PBMC treated with CPG ODN and Combination group exhibited more expression of Co-stimulatory molecules of CD80 and CD 86 than control group and Positive control group.
3. The effects of PBMC Stimulated by CPG ODN on hut-78 cells
The growth of hut-78 cells could be inhibited partially after being treated by PBMC activated by CPG ODN for 48h. The Cytotoxicity of control group, Positive control group, CPG ODN group, and Combination group was 24.93±7.81%, 41.86±2.93%,63.65±7.98%,67.05±3.56%.
4. The influence of PBMC stimulated by CPG ODN on cell morphology of hut-78 cells co-cultured PBMC
After co-cultured with PBMC stimulated by CpG ODN, morphological changes of hut-78 cells were more obvious:some cell boundaries unclear, and some cells shows nuclear enrichment.
Conclusions
1. CpG ODN could increase the secretions of INF-γand IL-12 of PBMC
2. CpG ODN could significantly exhibited more Co-stimulatory molecules of CD80 and CD 86.
3. CpG ODN could increase the cytotoxicity of PBMC on HUT-78 cells in vitro.
恶性淋巴瘤(Malignant Lymphoma, ML)是原发于淋巴结或淋巴结外组织或器官的一种恶性肿瘤,来源于淋巴细胞及其前体细胞。在我国恶性淋巴瘤虽相对少见,但近年来新发病例逐年上升,每年至少超过25000例,已成为严重威胁人类健康的一类疾病。恶性淋巴瘤的治疗以化疗为主,同时配合放疗和生物治疗等综合治疗。尽管一部分恶性淋巴瘤经过化疗等综合治疗后可以达到长期持续缓解,但总的来说恶性淋巴瘤因为其侵袭性、原发和继发耐药等原因,治疗效果往往不尽人意。因此,恶性淋巴瘤的攻克需要发展新的治疗途径与手段。目前研究较多的是肿瘤的免疫治疗,其中CpG ODN是肿瘤免疫治疗研究的热点。现已有多篇文献报道,CpG ODN对多种实体瘤的免疫治疗效果显著,但有关淋巴造血系统的恶性肿瘤研究较少。
过去几十年里大量的证据显示,病原体中的核酸不仅能作为生命的基本物质而存在,而且还可以激活宿主的免疫系统。细菌中未甲基化5'CpG3'二核苷酸(CpGoligodeoxynucleotide, CpG ODN)对各种免疫细胞亚群有强大的免疫激活作用。它们能直接激活巨噬细胞、单核细胞和树突状细胞等抗原递呈细胞,释放多种细胞因子,如白介素(IL)-12和IL-6等,再由这些细胞因子间接激活自然杀伤(NK)细胞及T细胞,发挥抗病毒、抗细菌感染及抗肿瘤作用。
本研究运用人工合成的含有CpG基序的寡聚脱氧核苷酸刺激人外周血单个核细胞(Peripheral blood mononuclear cell,PBMC),观察其对PBMC的活化刺激作用,以及由此介导的对淋巴瘤hut-78细胞的杀伤作用,旨在为淋巴瘤的免疫治疗提供一种新途径。
方法
淋巴细胞分离液分离健康人外周抗凝血,体外细胞培养技术常规培养PBMC；将实验分为四组：①阴性对照组：PBS+PBMC;②阳性对照组：LPS+PBMC；③实验组：CpG ODN+PBMC；④联合组：CpG ODN+LPS+PBMC.ELISA法检测CpG ODN活化PBMC后分泌IL-12.INF-γ的含量,流式细胞仪测活化PBMC表面标记CD80、CD86分子的表达；MTT法检测CpG ODN活化PBMC对hut-78细胞杀伤作用,倒置显微镜下观察细胞形态学变化。
结果
1 CpG ODN活化后PBMC培养液上清中IL-12和INF-γ的表达水平
CpG ODN活化后PBMC培养液上清中IL-12和INF-γ的表达水平显著升高。经LPS组、CpG ODN组及LPS+ CpG ODN组活化后的PBMC产生IL-12的含量分别60.54±1.82 pg.mL-1、115.50±19.34 pg.mL-1、120.75±13.71 pg.mL-1；产生INF-γ的含量分别为158.74±18.46 pg.mL-1、262.75±30.21 pg.mL-1、265.25±29.26 pg.mL-1。实验组结果与空白对照组差异有统计学意义(P<0.01)。
2 CpG ODN诱导PBMC表面标志的表达
与对照组相比,PBMC在经LPS组、CpG ODN组刺激后细胞表面标志协同刺激分子CD80、CD86的表达水平均明显增高(P<0.01)；CpG ODN组刺激的PBMC表达协同刺激分子CD80、CD86明显大于阳性对照LPS组(P<0.05)。
3 CpG ODN活化的PBMC对HUT-78细胞杀伤作用
CpG ODN活化后的PBMC对HUT-78细胞杀伤活力显著提高。经LPS组.CpG ODN组及CpG ODN+ LPS组活化后的人外周血单个核细胞对HUT-78细胞杀伤活力分别为41.86±2.93%、63.65±7.98%、67.05±3.56%。其实验组0D值与空白对照组差异有统计学意义(P<0.05)。
4形态学观察结果
通过观察发现HUT-78细胞与经CpG ODN刺激后的PBMC共同培养48小时后,形态学改变较明显：有些细胞边界不清,相互融合,部分细胞可见核浓缩。
结论
1经CpG ODN2006活化PBMC分泌细胞因子IL-12和INF-γ水平显著提高。
2 CpG ODN刺激PBMC表面表达的协同刺激分子CD80、CD86水平明显增高
3 CpG ODN显著增强人外周血单个核细胞对HUT-78细胞杀伤活力。
Background and Objective
Malignant lymphoma is a malignant proliferative disease generates from the human immune system cells and their precursor cells. Although malignant lymphoma in China is relatively rare, yet new cases have been increasing in recent years, at least more than 25,000 cases a year, which often occurs in young and seriously endangers the health and lives of human beings. Currently, combined chemotherapy is the common modality of the treatment of malignant lymphoma. It plays an important role in improving the curative rate and remarkably prolonging the disease-free interval of sufferers, but it has inevitably adverse reactions, such as bone marrow suppression and the decreasing of immune system, which make it difficult for the patients to complete the full course of treatment. Moreover, Primary or secondary drug resistance during treatment has become one of the tough problems of clinical therapy of the malignant tumor. Therefore, it is time to seek new ways and means of treatment for malignant lymphoma. More current researches are focus on immunotherapy of cancer and CpG ODN. A number of articles have been published that treatment with CpG ODN on solid tumors was significantly. However, the study of malignant tumors of lymphoid system is less.
DNAs in bacteria and virus, which contain numethylated CPG motifs activate host defense mechanisms leading to innate and acquired immune responses. Synthetic oligodeoxynucleotides(ODNs) containing unmethylated CPG motifs Minic the immunostimulatory qualities of bacterial DNA and elicit a coordinated set of T helper 1 (Thl)-like immune responses, including direct activation of B cells, plasmacytoid dendritic cells (PDCs), macrophages, antigen present cell (APC) and indirect activation of T cells, natural Killer (NK) cells. CpG ODN-induced activation of innate immunity protects against lethal challenge with a wide variety of pathogens. However, we can only find some reports on antitumor activity of CPG ODN in solid tumors.
So, in this study, we will investigate the antitumor effects of PBMC activated by Synthetic oligodeoxynucleotides (ODN) containing unmethylated CPG dinucleotides (CPG ODN) in vitro and explore the mechanisms of anti-tumor action of CPG ODN, which offers a new way for the treatment of the lymphoma.
Methods
Separating the circumference anti-hemoglutination of healthy persons with Lymphocyte separation medium, human peripheral blood mononuclear cells (PBMC) were cultured with technology of cell culture in vitro. We established four groups in the experiment:control group:PBMC+PBS; Positive control group:PBMC+LPS; ODN2006 group:PBMC+ODN2006; Combination group:PBMC+ODN2006+LPS. The secretions of INF-γand IL-12 from PBMC determined using ELISA method. After treated with these groups for 48 hours, Co-stimulatory molecules of CD80, CD86 on the surface of PBMC were analyzed by flow cytometry. PBMC activated by CPG ODN was co-cultured with hut-78 cells for 48 hours, and then MTT method was used to detect PBMC-mediated antitumor activity. We examined cell morphology with the inverted microscope.
Results
1. The effect of CpG ODN on INF-γand IL-12 of PBMC
The secretions of INF-γand IL-12 from PBMC treated with CpG ODN were increased significantly. The levers of IL-12 from PBMC of control group, Positive control group, CpG ODN group and Combination group was 15.50±10.12, 60.54±1.82 pg-mL-1,115.50±19.34 pg·mL-1,120.75±13.71 pg·mL-1,respectively; The levers of INF-γfrom PBMC of these four groups was 85.50±10.01, 158.74±18.46 pg·ml-1,262.75±30.21 pg·mL-1,265.25±29.26 pg·mL-1 separately.
2. The effect of PBMC treated with CpG ODN on Co-stimulatory molecules of CD80, CD86 of PBMC
PBMC treated with CPG ODN and Combination group exhibited more expression of Co-stimulatory molecules of CD80 and CD 86 than control group and Positive control group.
3. The effects of PBMC Stimulated by CPG ODN on hut-78 cells
The growth of hut-78 cells could be inhibited partially after being treated by PBMC activated by CPG ODN for 48h. The Cytotoxicity of control group, Positive control group, CPG ODN group, and Combination group was 24.93±7.81%, 41.86±2.93%,63.65±7.98%,67.05±3.56%.
4. The influence of PBMC stimulated by CPG ODN on cell morphology of hut-78 cells co-cultured PBMC
After co-cultured with PBMC stimulated by CpG ODN, morphological changes of hut-78 cells were more obvious:some cell boundaries unclear, and some cells shows nuclear enrichment.
Conclusions
1. CpG ODN could increase the secretions of INF-γand IL-12 of PBMC
2. CpG ODN could significantly exhibited more Co-stimulatory molecules of CD80 and CD 86.
3. CpG ODN could increase the cytotoxicity of PBMC on HUT-78 cells in vitro.
引文
1.孙燕,石远凯 主编.临床肿内科手册[M].第5版.北京:人民卫生出版社,2005,349.
2.潘启超.抗肿瘤药物的研究进展[J].癌症,1998,17(3):225-233.
3.刘学丽.CpG ODN的免疫活性及其在肿瘤治疗中的作用[J].中国肿瘤生物治疗杂志,2006,13(2):147-149.
4.Weiner GJ. CpG oligodeoxynucleotide-based therapy of lymphoid malignancies[J]. Adv Drag Deliv Rev. 2009,61 (3):263-267.
5.Krieg AM, Yi AK, Waldschmidt TJ,et al.CpG motifs in bacterial DNA trigger direct B-cell activation[J].Nature,1995,374(6522):546-549.
6.Krieg AM. Mechanisms and applications of immune stimulatory CpG oligodeoxynucleotides[J].Biochim Biophys Acta. 1999,1489(1):107-116.
8.Jahrsdorfer B, Weiner GJ. CpG oligodeoxynucleotides as immunotherapy in cancer[J]. Update Cancer Ther.2008,3(l):27-32.
9.Wang XS,Sheng Z,Ruan YB,et al.CpG oligodeoxynucleotides inhibit tumor growth and reverse the immunosuppression caused by me therapy with 5-fluorouracil in murine hepatoma [J].World J Gastroenterol,2005,11(8):1220-1224.
11.Wilson HL,Dar A,Napper SK,et al.Immune mechanisms and therapeutic potential of CpG oligodeoxynucleotides[J].Int Rev Immunol,2006,25 (3-4):183-213.
12.Tracey KJ, Cerami A. Tumor necrosis factor, other cytokines and disease[J].Annu Revcell Biol,1993 (9) :317-343.[10].
13.Wright SD. Toll, a new piece in the puzzle of innate immunity[J]. ExpMed,
14.颜伟,孙梯业.CpG ODNs的免疫活性作用与肿瘤治疗的研究进展[J].西南国防 医药,2006,16(4):455-461.
15.周捷,黄晓元,任利成.脆弱类杆菌脂多糖对正常人外周血单个核细胞分泌白介素的影响[J].2003,19(2):82-85.
16.郭乔楠,陈意生.白介素21抗肿瘤作用的进展[J].2020.9(3):353-35.5
17.Hemmi H, Takeuchi O, Kawai T, et al. A toll-like Receptor recognizes bacterial DNA, Nature, 2003,408(6813): 740-745.
18.Kline J N.Effect of CpG DNA on Th1/Th2 balance in asthma[J].Curr Top Microbiol Immunol,2000,24(7):211-225.
19.陈慰峰,主编.医学免疫学[M].第三版.北京:人民卫生出版社.2010.66-96
20.Hartmann G,Weeratna R D,Ballas Z K,et al. Delineation of a CpG phosphorothioate oligodeoxynucleotide for activating primary immune responses in vitro and in vivo[J].J Immunol,2000,16(4):1617-1624.
21.Mochizuki K, Hayashi N, Katayama K, et al. B7/BB-1 expression and hepatitis activity in liver tissues of patients with chronic hepatitis C [J]. Hepatology.1997,25(3):713-718.
22.曹庆伟,王彦军.共刺激分子B7在肿瘤免疫中的作用[J].国外医学外科学分册,2001,28(4):210-212.
23.Speiser DE, Lienard D, Rufer N, et al. Rapid and strong human CD8 + T cell responses to vaccination with peptide, IFA ,and CpG oligodeoxynucleotide 7909[J]. J Clin Invest, 2005,115(3) :739-746
24.Balsari A .Tortoreto M, Besusso D,et al .Combination of a CpG-oligodeoxynucleotide and a topoisom-erase Iinhibitor in the therapy of human tumour xenografts [J].Eur J Cancer,2004,40(8):1275-1281.
25.Wang XS,Sheng Z,Ruan YB.et al. CpG oligodeoxynucleotides inhibit tumor growth and reverse the immunosuppression caused by the therapy with 5-fluorouracil in murine hepatoma [J]. World J Gastroenterol ,2005,11(8):1220-1224.
26.Jahrsdorfer B, Weiner GJ, CpG oligodeoxynucleotides as immunotherapy in cancerCpG[J].Update Cancer Ther. 2008,3(1):27-32.
27. Forster R ,Ohl L, Henning G,et al. Chemokine receptor CCR7 directs dendritic cell differentiation ,mobilization and localization[A].In: Schuler G,Steinkasserer A ,Sting G.7th International Symposium on dendvitic cell[C], Germang, 2002,Auseria: Robidruck,2002:36.
28.Homung V ,Rothenfusser S,Britsch S ,et al. Quantitative expression of Toll-like receptor 1-10 mRNA in cellular subsets of human peripheral blood mononuclear cells and sensitivity to CpG oligodeoxynucleotides[J] J Immunol ,2002,168(9):4531-4537.
29.Londorf as,Kuekrek H.Stern BV,et al. Intratumor CpG oligodeoxynucleotide injection induces protective antitumor T cell immunity[J ]. J Immunol,2003,171(8):3941-3946.
30.Baines J,Celis E.Immune-mediated tumor regression induced by CpG-containing oligodeoxynucleotids[J].Clin Cancer Res,2003,9(7):2693-2700.
31.Jahrsdorfer B, Muhlenhoff L, Blackwell SE. et al. B-cell lymphomas differ in their responsiveness to CpG oligodeoxynucleotides[J]. Clin Cancer Res. 2005,11(4):1490-1499.
23.虞荣喜,罗秀素,毛建平.皮肤T细胞淋巴瘤三例报告[J].实用癌症杂志,1991,6(3):229-230.
33.韩晓凤,王燕婷,欧阳仁荣.MTT法体外药敏实验研究进展[J].肿瘤,1999,19(13):55.
34.Zimmennann S,Egeter O,Hausmann S,et al. CpG oligodeoxynucleotides trigger protective and curative Thl responses in lethal murine leishmaniasis[J].J Tmmoiinol 1998.160(8):3627-3630.
1. 陈洪亮,李伯涛,张家祥,等.免疫活性多糖的免疫调节作用及机制研究进展[J].中国药理学通报,2002,18(3)：249-252.
2.季宇彬,高世勇.多糖的研究进展[J].中草药,2003,34(增刊)：45-53.
3 Wilson HL,Dar A,Napper SK, et al.Immune mechanisms and therapeutic potential of Cp G oligodeoxynucleotides[J]. Int Rev Immunol,2006,25 (3-4):183-213.
4 Krieg AM. Therapeutic potential of Toll like receptor 9 activation[J]. Nat Rev Drug Discov.2006,5(6):471-484.
5 Klinman DM. Adjuvant activity of CpG oligodeoxynucleotides[J].Int Rev Immunol,2006,25 (3-4):135-154.
6 Wooldridge JE, Weiner GJ. CpG DNA and cancer immunotherapy:Orchestraing the antitumor immune response[J]. Curr Opin Oncol,2003,15(6):440-445
7 郭爱珍,陆承平.细菌CpG-DNA免疫激活活性进展[J].中国兽医学报,2002,22(3)：317～320.
8 王晓川,郭履.非甲基化胞嘧啶-腺嘌呤DNA的免疫调节作用[J].中华儿科杂志,1999,37(6)：65-69.
9 Krieg AM, Yi AK, Waldschmidt TJ,et al.CpG motifs in bacterial DNA trigger direct B-cell activation[J].Nature,1995,374(6522):546-549.
10 Yamamoto S, Yamamoto T, Iho S, et al. Activation of NK cell (human and mouse) by immunostimulatory DNA sequence. Springer Semin Immunopathol. 2000,22(1-2):35-43.
11陈晋.免疫刺激序列CpG的构效关系[J].中国兽医学报,1996,20(5)：350-353.
12 Krieg AM. Mechanisms and applications of immune stimulatory CpG oligodeoxynucleotides[J].Biochim Biophys Acta.1999,1489(1):107-116.
13 Yamamoto S,Yamamoto T, Shimada S,et al.DNA from bacterial,but not from Vertebrates, Induces interierons,Activates natural Killer Cells And Inhabits Tumor growth[J].Microbiol Immunol,1992,36(9):983-997.
14颜伟,孙梯业.CpG ODNs的免疫活性作用与肿瘤治疗的研究进展[J].西南国防医药,2006,16(4)：458-461.
15 Tracey KJ, Cerami A. Tumor necrosis factor, other cytokines and disease[J].Annu Revcell Biol,1993 (9):317-343.[10]
16 Wright SD. Toll, a new piece in the puzzle of innate immunity[J]. ExpMed, 1999,189 (2):605-609.
17熊杰.CpG寡核昔酸在免疫治疗中的作用[J].国际输血及血液学杂志,2006,29(6),525-528.
18 Hemmi H, Takeuchi O, Kawai T, et al. A toll-like Receptor recognizes bacterial DNA, Nature,2003,408(6813):740-745.
19刘学丽.CpG ODN的免疫活性及其在肿瘤治疗中的作用[J].中国肿瘤生物治疗杂志,2006,13(2)：147-149.
20 Bauer S,Kirschning CJ, Hacker H, et al.Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition[J]. Proc Natl Acad Sci U S A.2001,8(16):9237-9242.
21张秋玉.Toll样受体家族与TLRs信号转导[J].国外医学免疫学分册,2001,24(3)：142-146.
22魏伟,李晓辉.抗炎免疫药理学[M].北京：人民卫生出版社,2005：205-212.
23 Kisseleva T.Signaling through the JAK/STAT pathway:recent advances and future challenges[J]. Gene,2002,28(5):1121-1134
24 Marrero M B,Venema V J,He H.Inhibition by the JAK/STAI pathway of IFN gamma-and LPS stimulated nitricoxide synthase induction in vascular smooth muscle cells[J].Biochem Biophys ResCommun,2001,255(2):508-512.
25 Ajizian S J,English B K,Meals E A. Specific inhibitors of p38 and extracellular signal-regulated kinase mitogen-activated protein kinase pathways block inducible nitric oxide synthase and tumor necrosis factor accumulation in macrophages stimulated with lipopolysaccharide and inierferon-gamma[J].J Infect Dis,2001,181(5):939-944.
26 Davis H L.Use of CpG DNA for enhancing specific immune responses[J].Curr Top Microbiol Immunol,2000,24(7):171-181.
27 Kline J N.Effect of CpG DNA on Thl/Th2 balance in asthma[J].Curr Top Microbiol Immunol,2000,24(7):211-225.
28 Hartmann G,Weeratna R D,Ballas Z K,et al. Delineation of a CpG phosphorothioate oligodeoxynucleotide for activating primary immune responses in vitro and in vivo[J].J Immunol,2000,16(4):1617-1624.
29何威.新型的免疫佐剂：CpG寡聚脱氧核苷酸[J].中国临床医学,2008,15(5)：673-676.
30 Jahrsdorfer B, Weiner GJ, CpG oligodeoxynucleotides as immunotherapy in cancerCpG[J].Update Cancer Ther.2008,3(1):27-32.
31 Londorf as,Kuekrek H,Stern BV,et al. Intratumor CpG oligodeoxynucleotide injection induces protective antitumor T cell immunity[J]. J Immunol,2003,171 (8):3941-3946.
32 Baines J,Celis E.Immune-mediated tumor regression induced by CpG-containing oligodeoxynucleotids[J].Clin Cancer Res,2003,9(7):2693-2700.
33 Ballas ZK, Rasmussen WL, Krieg AM,et al.Induction of NK activity in murine and human cells by CpG motifs in oligodeoxynucleotides and bacterial DNA[J] Journal of Immunology.1996,157(5):1840-1845.
34 Stefan B, Carsterial J K, Hans H et al.Human TLR9 confera responsiveness to bacterial DNA via species-specific CpG motif recognition[J]. Proc Natl Acad Sci USA,2001,9(8):9237-9242.
35 Wooldridge JE,Ballas Z,Krieg AM, et al.Immunostimulatory oligodeoxynucleotides containing CpG motifs enhance the efficacy of monoclonal antibody therapy of lymphoma[J]. Blood.1997,89(8):2994-2998.
36 Ballas ZK,Krieg AM,Warren, T, et al. Divergent therapeutic and immunologic effects of oligodeoxynucleotides with distinct CpG motifs[J].J Immunol.2001,167(9):4878-4886.
37 Wang H,Rayburn ER, Wang W,et al.Immunomodulatory oligonucleotides as novel therapy for breast cancer:pharmacokinetics, in vitro and in vivo anticancer activity, and potentiation of antibody therapy[J].Mol Cancer Ther. 2006,5(8):2106-2114.
38 Leonard JP,Link BK,Emmanouilides, C,et al. Phase I Trial of Toll-Like Receptor 9 Agonist PF-3512676 with and Following Rituximab in Patients with Recurrent Indolent and Aggressive Non Hodgkin's Lymphoma. Clin Cancer Res. 2007,13(20):6168-6174
39 Jahrsdorfer B,Muhlenhoff L,Blackwell SE,et al.B-cell lymphomas differ in their responsiveness to CpG oligodeoxynucleotides[J].Clin Cancer Res. 2005,11(4):1490-1499.
40 Friedberg JW,Kim H, McCauley M,et al.Combination immunotherapy with a CpG oligonucleotide (1018 ISS) and rituximab in patients with non-Hodgkin lymphoma:increased interferon-alpha/beta-inducible gene expression, without significant toxicity[J]. Blood.2005,105(2):489-495.
41 Weigel BJ,Rodeberg DA,Krieg AM, et al.CpG oligodeoxynucleotides potentiate the antitumor effects of chemotherapy or tumor resection in an orthotopic murine model of rhabdomyosarcoma[J]. Clin Cancer Res.2003,9(8):3105-3114.
42 Pratesi G, Petrangolini G, Tortoreto, M,et al.Therapeutic synergism of gemcitabine and CpG-oligodeoxynucleotides in an orthotopic human pancreatic carcinoma xenograft[J]. Cancer Res.2005,65(14):6388-6393.
43 vanderMost RG, Himbeck R, Aarons S,et al. Antitumor efficacy of the novel chemotherapeutic agent coramsine is potentiated by cotreatment with CpG-containing oligodeoxynucleotides[J].J.Immunother.2006,29(2):134-142.
44 Manegold C, Gravenor D, Woytowitz D,et al. Randomized phase II trial of a toll-like receptor 9 agonist oligodeoxynucleotide, PF-3512676, in combination with first-line taxane plus platinum chemotherapy for advanced-stage non-small-cell lung cancer[J]. J Clin Oncol.2008,6(24):3979-3986.
45 Klinman DM, Xie H, Ivins BE.CpG oligonucleotides improve the protective immune response induced by the licensed anthrax vaccine[J]. Ann N YAcad Sci.2006,137-150.
46 Cooper CL, Davis HL, Morris ML,et al. CPG 7909, an immunostimulatory TLR9 agonist oligodeoxynucleotide, as adjuvant to Engerix-B HB V vaccine in healthy adults:a double-blind phase Ⅰ/Ⅱ study[J].J Clin Immunol.2004,24(6):693-701.
47 Weiner GJ,Liu HM, Wooldridge JE,et al.Immunostimulatory Oligodeoxynucleotides Containing the CpG Motif are Effective As Immune Adjuvants In Tumor Antigen Immunization[J].Proceedings of the National Academy of Sciences of the United States of America. 1997,94(20):10833-10837.
48 Gendron KB,Rodriguez A, Sewell DA.Vaccination with human papillomavirus type 16 E7 peptide with CpG oligonucleotides for prevention of tumor growth in mice[J]. Arch Otolaryngol Head Neck Surg.2006,132(3):327-332.
49 Mukherjee P, Pathangey LB, Bradley JB,et al.MUC1-specific immune therapy generates a strong anti-tumor response in a MUC1-tolerant colon cancer model[J]. Vaccine.2007,25(9):1607-1618.
50 Kim SK, Ragupathi G, Cappello S,et al. Effect of immunological adjuvant combinations on the antibody and T-cell response to vaccination with MUC1-KLH and GD3-KLH conjugates[J]. Vaccine.2000,19(4-5):530-537.
51 Ohashi K, Kobayashi G, Fang S,et al.Surgical excision combined with autologous whole tumor cell vaccination is an effective therapy for murine neuroblastoma. J Pediatr Surg.2006,41(8):1361-1368.
52 Pilon-Thomas S, Li W, Briggs JJ,et al. Immunostimulatory effects of CpG-ODN upon dendritic cell-based immunotherapy in a murine melanoma model[J]. J Immunother.2006,29(4):381-387.
53 Du YC, Lin P, Zhang J, et al.Fusion of CpG-ODN-stimulating dendritic cells with Lewis lung cancer cells can enhance anti-tumor immune responses[J]. Tissue Antigens.2006,67(5):368-376.
54 Brown L, Roda J, Terrell C,et al.Interferon alpha and CPG oligodeoxynucleotides elicit additive immunostimulatory and antitumor effects[J].Surgery. 2006,140(2):297-306.
55 Chaudhry UI, Kingham TP, Plitas G,et al.Combined stimulation with interleukin-18 and CpG induces murine natural killer dendritic cells to produce IFN-gamma and inhibit tumor growth[J]. Cancer Res. 2006,66(21):10497-10504.
56 Chuang CM, Monie A, Wu A,et al.Treatment with LL-37 Peptide Enhances Antitumor Effects Induced by CpG Oligodeoxynucleotides Against Ovarian Cancer[J]. Hum Gene Ther.2009,20(4):303-313.
57 Mason KA,Neal R,Hunter N,et al.CpG oligodeoxynucleotides are potent enhancers of radio-and chemoresponses of murine tumors[J]. Radiother Oncol 2006,80(2):192-198.
58 Meng Y, Carpentier AF, Chen L,et al. Successful combination of local CpG-ODN and radiotherapy in malignant glioma[J]. Int J Cancer 2005,116(6):992-997.
59 Ai WYZ, Kim Y, Hoppe RT,et al. Preliminary Report on a Phase I/II Study of Intratumoral Injection of PF-3512676 (CpG 7909), a TLR9 Agonist, Combined with Radiation in Recurrent Low-Grade Lymphomas[J]. Blood 2006,108(11)Abstract #2716
60 Brok MH, Sutmuller RP, Nierkens S,et al. Synergy between in situ cryoablation and TLR9 stimulation results in a highly effective in vivo dendritic cell vaccine[J]. Cancer Res 2006,66(14):7285-7292.
61 Chamoto K, Takeshima T, Wakita D,et al. Combination immunotherapy with radiation and CpG-based tumor vaccination for the eradication of radio-and immuno-resistant lung carcinoma cells[J]. Cancer Sci.009,100(5):934-939.
62 Roux S, Bernat C, Al-Sakere B,et al. Tumor destruction using electrochemotherapy followed by CpG oligodeoxynucleotide injection induces distant tumor responses[J]. Cancer Immunol Immunother.2008,57(9):1291-300.
63 Wingender G, Garbi N, Schumak B, et al. Systemic application of Cp G-rich DNA suppresses adaptive T cell immunity via induction of IDO[J]. Eur J Immunol,2006,36 (1):12-20.
64 Krieg AM, Efler SM, Wittpoth M, et al. Induction of systemic TH1-like innate immunity in normal volunteers following subcutaneous but not int ravenous administ ration of CPG 7909, asynthetic B-class CpG oligodeoxynucleotide TLR9 agonist [J]. J Immunother,2004,27 (6):460-471
65 Heikenwalder M, Poiymenidou M, Junt T, et al. Lymphoid follicle descfuction and immunosuppression after repeated CpG oligodcoxynucleotide administration. Nat Med,2004,10(2):187-192
66 Klinman DM, Takeno M, Ichino M, et al. DNA vaccines:safety and efficacy issues [J]. Springer Semin Immunopathol,1997,19 (2):245-256.
2.潘启超.抗肿瘤药物的研究进展[J].癌症,1998,17(3):225-233.
3.刘学丽.CpG ODN的免疫活性及其在肿瘤治疗中的作用[J].中国肿瘤生物治疗杂志,2006,13(2):147-149.
4.Weiner GJ. CpG oligodeoxynucleotide-based therapy of lymphoid malignancies[J]. Adv Drag Deliv Rev. 2009,61 (3):263-267.
5.Krieg AM, Yi AK, Waldschmidt TJ,et al.CpG motifs in bacterial DNA trigger direct B-cell activation[J].Nature,1995,374(6522):546-549.
6.Krieg AM. Mechanisms and applications of immune stimulatory CpG oligodeoxynucleotides[J].Biochim Biophys Acta. 1999,1489(1):107-116.
8.Jahrsdorfer B, Weiner GJ. CpG oligodeoxynucleotides as immunotherapy in cancer[J]. Update Cancer Ther.2008,3(l):27-32.
9.Wang XS,Sheng Z,Ruan YB,et al.CpG oligodeoxynucleotides inhibit tumor growth and reverse the immunosuppression caused by me therapy with 5-fluorouracil in murine hepatoma [J].World J Gastroenterol,2005,11(8):1220-1224.
11.Wilson HL,Dar A,Napper SK,et al.Immune mechanisms and therapeutic potential of CpG oligodeoxynucleotides[J].Int Rev Immunol,2006,25 (3-4):183-213.
12.Tracey KJ, Cerami A. Tumor necrosis factor, other cytokines and disease[J].Annu Revcell Biol,1993 (9) :317-343.[10].
13.Wright SD. Toll, a new piece in the puzzle of innate immunity[J]. ExpMed,
14.颜伟,孙梯业.CpG ODNs的免疫活性作用与肿瘤治疗的研究进展[J].西南国防 医药,2006,16(4):455-461.
15.周捷,黄晓元,任利成.脆弱类杆菌脂多糖对正常人外周血单个核细胞分泌白介素的影响[J].2003,19(2):82-85.
16.郭乔楠,陈意生.白介素21抗肿瘤作用的进展[J].2020.9(3):353-35.5
17.Hemmi H, Takeuchi O, Kawai T, et al. A toll-like Receptor recognizes bacterial DNA, Nature, 2003,408(6813): 740-745.
18.Kline J N.Effect of CpG DNA on Th1/Th2 balance in asthma[J].Curr Top Microbiol Immunol,2000,24(7):211-225.
19.陈慰峰,主编.医学免疫学[M].第三版.北京:人民卫生出版社.2010.66-96
20.Hartmann G,Weeratna R D,Ballas Z K,et al. Delineation of a CpG phosphorothioate oligodeoxynucleotide for activating primary immune responses in vitro and in vivo[J].J Immunol,2000,16(4):1617-1624.
21.Mochizuki K, Hayashi N, Katayama K, et al. B7/BB-1 expression and hepatitis activity in liver tissues of patients with chronic hepatitis C [J]. Hepatology.1997,25(3):713-718.
22.曹庆伟,王彦军.共刺激分子B7在肿瘤免疫中的作用[J].国外医学外科学分册,2001,28(4):210-212.
23.Speiser DE, Lienard D, Rufer N, et al. Rapid and strong human CD8 + T cell responses to vaccination with peptide, IFA ,and CpG oligodeoxynucleotide 7909[J]. J Clin Invest, 2005,115(3) :739-746
24.Balsari A .Tortoreto M, Besusso D,et al .Combination of a CpG-oligodeoxynucleotide and a topoisom-erase Iinhibitor in the therapy of human tumour xenografts [J].Eur J Cancer,2004,40(8):1275-1281.
25.Wang XS,Sheng Z,Ruan YB.et al. CpG oligodeoxynucleotides inhibit tumor growth and reverse the immunosuppression caused by the therapy with 5-fluorouracil in murine hepatoma [J]. World J Gastroenterol ,2005,11(8):1220-1224.
26.Jahrsdorfer B, Weiner GJ, CpG oligodeoxynucleotides as immunotherapy in cancerCpG[J].Update Cancer Ther. 2008,3(1):27-32.
27. Forster R ,Ohl L, Henning G,et al. Chemokine receptor CCR7 directs dendritic cell differentiation ,mobilization and localization[A].In: Schuler G,Steinkasserer A ,Sting G.7th International Symposium on dendvitic cell[C], Germang, 2002,Auseria: Robidruck,2002:36.
28.Homung V ,Rothenfusser S,Britsch S ,et al. Quantitative expression of Toll-like receptor 1-10 mRNA in cellular subsets of human peripheral blood mononuclear cells and sensitivity to CpG oligodeoxynucleotides[J] J Immunol ,2002,168(9):4531-4537.
29.Londorf as,Kuekrek H.Stern BV,et al. Intratumor CpG oligodeoxynucleotide injection induces protective antitumor T cell immunity[J ]. J Immunol,2003,171(8):3941-3946.
30.Baines J,Celis E.Immune-mediated tumor regression induced by CpG-containing oligodeoxynucleotids[J].Clin Cancer Res,2003,9(7):2693-2700.
31.Jahrsdorfer B, Muhlenhoff L, Blackwell SE. et al. B-cell lymphomas differ in their responsiveness to CpG oligodeoxynucleotides[J]. Clin Cancer Res. 2005,11(4):1490-1499.
23.虞荣喜,罗秀素,毛建平.皮肤T细胞淋巴瘤三例报告[J].实用癌症杂志,1991,6(3):229-230.
33.韩晓凤,王燕婷,欧阳仁荣.MTT法体外药敏实验研究进展[J].肿瘤,1999,19(13):55.
34.Zimmennann S,Egeter O,Hausmann S,et al. CpG oligodeoxynucleotides trigger protective and curative Thl responses in lethal murine leishmaniasis[J].J Tmmoiinol 1998.160(8):3627-3630.
1. 陈洪亮,李伯涛,张家祥,等.免疫活性多糖的免疫调节作用及机制研究进展[J].中国药理学通报,2002,18(3)：249-252.
2.季宇彬,高世勇.多糖的研究进展[J].中草药,2003,34(增刊)：45-53.
3 Wilson HL,Dar A,Napper SK, et al.Immune mechanisms and therapeutic potential of Cp G oligodeoxynucleotides[J]. Int Rev Immunol,2006,25 (3-4):183-213.
4 Krieg AM. Therapeutic potential of Toll like receptor 9 activation[J]. Nat Rev Drug Discov.2006,5(6):471-484.
5 Klinman DM. Adjuvant activity of CpG oligodeoxynucleotides[J].Int Rev Immunol,2006,25 (3-4):135-154.
6 Wooldridge JE, Weiner GJ. CpG DNA and cancer immunotherapy:Orchestraing the antitumor immune response[J]. Curr Opin Oncol,2003,15(6):440-445
7 郭爱珍,陆承平.细菌CpG-DNA免疫激活活性进展[J].中国兽医学报,2002,22(3)：317～320.
8 王晓川,郭履.非甲基化胞嘧啶-腺嘌呤DNA的免疫调节作用[J].中华儿科杂志,1999,37(6)：65-69.
9 Krieg AM, Yi AK, Waldschmidt TJ,et al.CpG motifs in bacterial DNA trigger direct B-cell activation[J].Nature,1995,374(6522):546-549.
10 Yamamoto S, Yamamoto T, Iho S, et al. Activation of NK cell (human and mouse) by immunostimulatory DNA sequence. Springer Semin Immunopathol. 2000,22(1-2):35-43.
11陈晋.免疫刺激序列CpG的构效关系[J].中国兽医学报,1996,20(5)：350-353.
12 Krieg AM. Mechanisms and applications of immune stimulatory CpG oligodeoxynucleotides[J].Biochim Biophys Acta.1999,1489(1):107-116.
13 Yamamoto S,Yamamoto T, Shimada S,et al.DNA from bacterial,but not from Vertebrates, Induces interierons,Activates natural Killer Cells And Inhabits Tumor growth[J].Microbiol Immunol,1992,36(9):983-997.
14颜伟,孙梯业.CpG ODNs的免疫活性作用与肿瘤治疗的研究进展[J].西南国防医药,2006,16(4)：458-461.
15 Tracey KJ, Cerami A. Tumor necrosis factor, other cytokines and disease[J].Annu Revcell Biol,1993 (9):317-343.[10]
16 Wright SD. Toll, a new piece in the puzzle of innate immunity[J]. ExpMed, 1999,189 (2):605-609.
17熊杰.CpG寡核昔酸在免疫治疗中的作用[J].国际输血及血液学杂志,2006,29(6),525-528.
18 Hemmi H, Takeuchi O, Kawai T, et al. A toll-like Receptor recognizes bacterial DNA, Nature,2003,408(6813):740-745.
19刘学丽.CpG ODN的免疫活性及其在肿瘤治疗中的作用[J].中国肿瘤生物治疗杂志,2006,13(2)：147-149.
20 Bauer S,Kirschning CJ, Hacker H, et al.Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition[J]. Proc Natl Acad Sci U S A.2001,8(16):9237-9242.
21张秋玉.Toll样受体家族与TLRs信号转导[J].国外医学免疫学分册,2001,24(3)：142-146.
22魏伟,李晓辉.抗炎免疫药理学[M].北京：人民卫生出版社,2005：205-212.
23 Kisseleva T.Signaling through the JAK/STAT pathway:recent advances and future challenges[J]. Gene,2002,28(5):1121-1134
24 Marrero M B,Venema V J,He H.Inhibition by the JAK/STAI pathway of IFN gamma-and LPS stimulated nitricoxide synthase induction in vascular smooth muscle cells[J].Biochem Biophys ResCommun,2001,255(2):508-512.
25 Ajizian S J,English B K,Meals E A. Specific inhibitors of p38 and extracellular signal-regulated kinase mitogen-activated protein kinase pathways block inducible nitric oxide synthase and tumor necrosis factor accumulation in macrophages stimulated with lipopolysaccharide and inierferon-gamma[J].J Infect Dis,2001,181(5):939-944.
26 Davis H L.Use of CpG DNA for enhancing specific immune responses[J].Curr Top Microbiol Immunol,2000,24(7):171-181.
27 Kline J N.Effect of CpG DNA on Thl/Th2 balance in asthma[J].Curr Top Microbiol Immunol,2000,24(7):211-225.
28 Hartmann G,Weeratna R D,Ballas Z K,et al. Delineation of a CpG phosphorothioate oligodeoxynucleotide for activating primary immune responses in vitro and in vivo[J].J Immunol,2000,16(4):1617-1624.
29何威.新型的免疫佐剂：CpG寡聚脱氧核苷酸[J].中国临床医学,2008,15(5)：673-676.
30 Jahrsdorfer B, Weiner GJ, CpG oligodeoxynucleotides as immunotherapy in cancerCpG[J].Update Cancer Ther.2008,3(1):27-32.
31 Londorf as,Kuekrek H,Stern BV,et al. Intratumor CpG oligodeoxynucleotide injection induces protective antitumor T cell immunity[J]. J Immunol,2003,171 (8):3941-3946.
32 Baines J,Celis E.Immune-mediated tumor regression induced by CpG-containing oligodeoxynucleotids[J].Clin Cancer Res,2003,9(7):2693-2700.
33 Ballas ZK, Rasmussen WL, Krieg AM,et al.Induction of NK activity in murine and human cells by CpG motifs in oligodeoxynucleotides and bacterial DNA[J] Journal of Immunology.1996,157(5):1840-1845.
34 Stefan B, Carsterial J K, Hans H et al.Human TLR9 confera responsiveness to bacterial DNA via species-specific CpG motif recognition[J]. Proc Natl Acad Sci USA,2001,9(8):9237-9242.
35 Wooldridge JE,Ballas Z,Krieg AM, et al.Immunostimulatory oligodeoxynucleotides containing CpG motifs enhance the efficacy of monoclonal antibody therapy of lymphoma[J]. Blood.1997,89(8):2994-2998.
36 Ballas ZK,Krieg AM,Warren, T, et al. Divergent therapeutic and immunologic effects of oligodeoxynucleotides with distinct CpG motifs[J].J Immunol.2001,167(9):4878-4886.
37 Wang H,Rayburn ER, Wang W,et al.Immunomodulatory oligonucleotides as novel therapy for breast cancer:pharmacokinetics, in vitro and in vivo anticancer activity, and potentiation of antibody therapy[J].Mol Cancer Ther. 2006,5(8):2106-2114.
38 Leonard JP,Link BK,Emmanouilides, C,et al. Phase I Trial of Toll-Like Receptor 9 Agonist PF-3512676 with and Following Rituximab in Patients with Recurrent Indolent and Aggressive Non Hodgkin's Lymphoma. Clin Cancer Res. 2007,13(20):6168-6174
39 Jahrsdorfer B,Muhlenhoff L,Blackwell SE,et al.B-cell lymphomas differ in their responsiveness to CpG oligodeoxynucleotides[J].Clin Cancer Res. 2005,11(4):1490-1499.
40 Friedberg JW,Kim H, McCauley M,et al.Combination immunotherapy with a CpG oligonucleotide (1018 ISS) and rituximab in patients with non-Hodgkin lymphoma:increased interferon-alpha/beta-inducible gene expression, without significant toxicity[J]. Blood.2005,105(2):489-495.
41 Weigel BJ,Rodeberg DA,Krieg AM, et al.CpG oligodeoxynucleotides potentiate the antitumor effects of chemotherapy or tumor resection in an orthotopic murine model of rhabdomyosarcoma[J]. Clin Cancer Res.2003,9(8):3105-3114.
42 Pratesi G, Petrangolini G, Tortoreto, M,et al.Therapeutic synergism of gemcitabine and CpG-oligodeoxynucleotides in an orthotopic human pancreatic carcinoma xenograft[J]. Cancer Res.2005,65(14):6388-6393.
43 vanderMost RG, Himbeck R, Aarons S,et al. Antitumor efficacy of the novel chemotherapeutic agent coramsine is potentiated by cotreatment with CpG-containing oligodeoxynucleotides[J].J.Immunother.2006,29(2):134-142.
44 Manegold C, Gravenor D, Woytowitz D,et al. Randomized phase II trial of a toll-like receptor 9 agonist oligodeoxynucleotide, PF-3512676, in combination with first-line taxane plus platinum chemotherapy for advanced-stage non-small-cell lung cancer[J]. J Clin Oncol.2008,6(24):3979-3986.
45 Klinman DM, Xie H, Ivins BE.CpG oligonucleotides improve the protective immune response induced by the licensed anthrax vaccine[J]. Ann N YAcad Sci.2006,137-150.
46 Cooper CL, Davis HL, Morris ML,et al. CPG 7909, an immunostimulatory TLR9 agonist oligodeoxynucleotide, as adjuvant to Engerix-B HB V vaccine in healthy adults:a double-blind phase Ⅰ/Ⅱ study[J].J Clin Immunol.2004,24(6):693-701.
47 Weiner GJ,Liu HM, Wooldridge JE,et al.Immunostimulatory Oligodeoxynucleotides Containing the CpG Motif are Effective As Immune Adjuvants In Tumor Antigen Immunization[J].Proceedings of the National Academy of Sciences of the United States of America. 1997,94(20):10833-10837.
48 Gendron KB,Rodriguez A, Sewell DA.Vaccination with human papillomavirus type 16 E7 peptide with CpG oligonucleotides for prevention of tumor growth in mice[J]. Arch Otolaryngol Head Neck Surg.2006,132(3):327-332.
49 Mukherjee P, Pathangey LB, Bradley JB,et al.MUC1-specific immune therapy generates a strong anti-tumor response in a MUC1-tolerant colon cancer model[J]. Vaccine.2007,25(9):1607-1618.
50 Kim SK, Ragupathi G, Cappello S,et al. Effect of immunological adjuvant combinations on the antibody and T-cell response to vaccination with MUC1-KLH and GD3-KLH conjugates[J]. Vaccine.2000,19(4-5):530-537.
51 Ohashi K, Kobayashi G, Fang S,et al.Surgical excision combined with autologous whole tumor cell vaccination is an effective therapy for murine neuroblastoma. J Pediatr Surg.2006,41(8):1361-1368.
52 Pilon-Thomas S, Li W, Briggs JJ,et al. Immunostimulatory effects of CpG-ODN upon dendritic cell-based immunotherapy in a murine melanoma model[J]. J Immunother.2006,29(4):381-387.
53 Du YC, Lin P, Zhang J, et al.Fusion of CpG-ODN-stimulating dendritic cells with Lewis lung cancer cells can enhance anti-tumor immune responses[J]. Tissue Antigens.2006,67(5):368-376.
54 Brown L, Roda J, Terrell C,et al.Interferon alpha and CPG oligodeoxynucleotides elicit additive immunostimulatory and antitumor effects[J].Surgery. 2006,140(2):297-306.
55 Chaudhry UI, Kingham TP, Plitas G,et al.Combined stimulation with interleukin-18 and CpG induces murine natural killer dendritic cells to produce IFN-gamma and inhibit tumor growth[J]. Cancer Res. 2006,66(21):10497-10504.
56 Chuang CM, Monie A, Wu A,et al.Treatment with LL-37 Peptide Enhances Antitumor Effects Induced by CpG Oligodeoxynucleotides Against Ovarian Cancer[J]. Hum Gene Ther.2009,20(4):303-313.
57 Mason KA,Neal R,Hunter N,et al.CpG oligodeoxynucleotides are potent enhancers of radio-and chemoresponses of murine tumors[J]. Radiother Oncol 2006,80(2):192-198.
58 Meng Y, Carpentier AF, Chen L,et al. Successful combination of local CpG-ODN and radiotherapy in malignant glioma[J]. Int J Cancer 2005,116(6):992-997.
59 Ai WYZ, Kim Y, Hoppe RT,et al. Preliminary Report on a Phase I/II Study of Intratumoral Injection of PF-3512676 (CpG 7909), a TLR9 Agonist, Combined with Radiation in Recurrent Low-Grade Lymphomas[J]. Blood 2006,108(11)Abstract #2716
60 Brok MH, Sutmuller RP, Nierkens S,et al. Synergy between in situ cryoablation and TLR9 stimulation results in a highly effective in vivo dendritic cell vaccine[J]. Cancer Res 2006,66(14):7285-7292.
61 Chamoto K, Takeshima T, Wakita D,et al. Combination immunotherapy with radiation and CpG-based tumor vaccination for the eradication of radio-and immuno-resistant lung carcinoma cells[J]. Cancer Sci.009,100(5):934-939.
62 Roux S, Bernat C, Al-Sakere B,et al. Tumor destruction using electrochemotherapy followed by CpG oligodeoxynucleotide injection induces distant tumor responses[J]. Cancer Immunol Immunother.2008,57(9):1291-300.
63 Wingender G, Garbi N, Schumak B, et al. Systemic application of Cp G-rich DNA suppresses adaptive T cell immunity via induction of IDO[J]. Eur J Immunol,2006,36 (1):12-20.
64 Krieg AM, Efler SM, Wittpoth M, et al. Induction of systemic TH1-like innate immunity in normal volunteers following subcutaneous but not int ravenous administ ration of CPG 7909, asynthetic B-class CpG oligodeoxynucleotide TLR9 agonist [J]. J Immunother,2004,27 (6):460-471
65 Heikenwalder M, Poiymenidou M, Junt T, et al. Lymphoid follicle descfuction and immunosuppression after repeated CpG oligodcoxynucleotide administration. Nat Med,2004,10(2):187-192
66 Klinman DM, Takeno M, Ichino M, et al. DNA vaccines:safety and efficacy issues [J]. Springer Semin Immunopathol,1997,19 (2):245-256.