mB7.1-GPI膜表面修饰的结肠癌细胞疫苗的抗肿瘤作用研究

目的制备mB7.1-GPI膜表面修饰的结肠癌细胞疫苗,并探讨其抗肿瘤作用。方法将mB7.1-GPI融合蛋白锚定于小鼠结肠癌细胞膜上,制备结肠癌细胞疫苗,并采用流式细胞术检测该蛋白的细胞膜锚定效果;建立C57BL/6小鼠结肠癌模型,检测mB7.1-GPI融合蛋白肿瘤疫苗对荷瘤小鼠的免疫治疗作用。结果 mB7.1-GPI融合蛋白可锚定在小鼠结肠癌细胞膜上,并有效刺激小鼠脾细胞增殖和分泌IL-2和IFN-γ,mB7.1-GPI膜表面修饰的结肠癌细胞疫苗能够显著抑制荷瘤小鼠肿瘤的生长,并延长其生存期。结论 mB7.1-GPI融合蛋白制备的结肠癌细胞疫苗具有较强的抗肿瘤作用,可能成为一种有效的、用于结肠癌的治疗和预防的新型疫苗。     

水溶性壳聚糖抗肿瘤作用的实验研究

目的研究水溶性壳聚糖对抗肿瘤活性因子和NK细胞活性的影响。方法分别用不同浓度水溶性壳聚糖腹腔注射荷瘤小鼠 ,然后测定巨噬细胞一氧化氮 (NO)、肿瘤坏死因子 (TNF)的生成 ,以及脾细胞γ 干扰素(IFN γ)和NK细胞活性。结果水溶性壳聚糖有显著地促进荷瘤小鼠NO、TNF、IFN γ的生成 ,提高NK细胞活性。与对照相比P <0 .0 1。结论水溶性壳聚糖能提高荷瘤小鼠的免疫功能。     

榄香烯哌嗪对荷瘤小鼠免疫功能的影响

目的研究榄香烯哌嗪的抗肿瘤作用及其对免疫功能的影响。方法采用小鼠S180肉瘤移植性肿瘤动物模型,以抑瘤率为指标考察榄香烯哌嗪的体内抗肿瘤活性。并用四氮唑盐(MTT)法对荷瘤小鼠进行脾淋巴细胞增殖能力和自然杀伤(NK)细胞活性测定,考察其对荷瘤小鼠免疫功能的影响。结果榄香烯哌嗪对小鼠肉瘤S180的生长有明显的抑制作用(P<0.01),502、5 mg.kg-1剂量组抑瘤率分别为48.74%和37.60%。榄香烯哌嗪各种剂量对荷瘤小鼠的胸腺指数和脾指数无明显影响,但榄香烯哌嗪剂量为502、5 mg.kg-1可显著提高荷瘤小鼠脾淋巴细胞的增殖能力,对荷瘤小鼠NK细胞活性也有明显的提高作用。结论榄香烯哌嗪具有一定的抗肿瘤作用,其抗肿瘤作用可能与激活体内免疫系统有关。     

螺旋藻多糖硫酸酯化修饰前后抗肿瘤及免疫活性的研究

比较螺旋藻多糖硫酸酯化修饰前后体内外抗肿瘤及免疫活性.采用MTT比色法研究药物在体外对人肿瘤细胞株的抑制作用,促进正常小鼠脾淋巴细胞的增殖活性以及对荷瘤小鼠NK和CTL细胞活性的影响.采用移植性肿瘤实验方法考察了药物对小鼠S180肉瘤的抑制作用.结果显示,螺旋藻多糖（NPSP）对肿瘤细胞株几乎无细胞毒作用,硫酸酯化螺旋藻多糖（SNPSP）对肿瘤细胞株具有显著的细胞毒作用,其中对SMMC-7721人肝癌细胞株抑制率最高达50%.NPSP50 mg/kg对小鼠S180肉瘤无抑制,相同剂量的SNPSP对小鼠S180肉瘤的抑制率达35.42%.NPSP具有促进脾淋巴细胞增殖作用,但对ConA和LPS诱导的脾淋巴细胞增殖反应无促进作用,SNPSP促进脾淋巴细胞增殖作用较NPSP增强,同时对ConA和LPS诱导的脾淋巴细胞增殖反应也具有明显的促进作用.NPSP和SNPSP均能促进荷瘤小鼠NK细胞和CTL细胞活性,其中SNPSP促进CTL细胞活性较NPSP增强.     

绞股蓝总皂甙对Lewis肺癌荷瘤小鼠肿瘤生长及免疫功能的影响

目的 :观察绞股蓝总皂甙 (gypenosides,GPs)对Lewis肺癌荷瘤小鼠肿瘤生长、脾淋巴细胞数及NK活性的影响。方法 :整体动物的抑瘤试验、脾淋巴细胞计数及NK活性测定。结果 :GPs对荷瘤小鼠Lewis肺癌细胞具有明显的抑制作用 ,在剂量 1 0、2 0、4 0mg/kg腹腔注射 (ip)给药条件下 ,其抑瘤率分别为 (3 0 7± 1 2 ) % ,(51 5± 2 5) %和 (50 3± 1 5) % (P <0 0 1 )。同时 ,GPsip给药后荷瘤小鼠脾淋巴细胞总数明显增加 ,外周血淋巴细胞NK活性明显升高 ,脾淋巴细胞NK活性也明显升高。结论 :GPs的抗肿瘤作用与其对荷瘤小鼠免疫功能的增强作用密切相关。     

致敏树突状细胞疫苗抗小鼠乳腺癌的实验研究

目的研究以EMT6乳腺癌细胞致敏的树突状细胞(DC)疫苗的抗肿瘤作用。方法无菌取小鼠骨髓细胞,在体外培养条件下经细胞因子诱导为DC,用EMT6肿瘤细胞冻融抗原冲击致敏DC,检测经DC免疫产生的细胞毒T淋巴细胞(CTL)体外杀伤肿瘤细胞的活性。建立EMT6荷瘤小鼠模型,随机分为实验组、实验对照组和空白对照组,于肿瘤接种后第7、14天给予相应DC疫苗治疗,观察致敏DC免疫对小鼠乳腺肿瘤模型的治疗作用。结果致敏DC诱导生成的特异性CTL在体外对肿瘤细胞可产生杀伤作用,与PBS对照组比较,差异有显著性(P<0.05);经致敏DC注射免疫后,小鼠移植瘤得到抑制,与PBS对照组比较,差异有显著性(P<0.05)。结论以EMT6乳腺癌细胞致敏的树突状细胞(DC)疫苗在体外和小鼠体内均显示了抗肿瘤作用。     

以链球菌制剂OK432为佐剂的内皮细胞疫苗体内抗小鼠肝癌转移的作用（英文）

目的评估OK432作为佐剂制备的内皮细胞疫苗体内抗肿瘤转移的作用。方法以OK432为佐剂,体外混合人脐静脉内皮细胞(HUVEC),制备HUVEC-OK432疫苗。雄性健康BALB/c小鼠尾静脉注射肝癌细胞5×105建立肝癌肺转移模型,皮下注射HUVEC-OK432疫苗,分别在预防性和治疗性免疫中评价HUVEC-OK432疫苗抗肿瘤转移活性;皮内注射5×104肝癌细胞建立皮内肿瘤血管模型,皮下注射HUVEC-OK432疫苗,评价HUVEC-OK432疫苗抑制肿瘤新生血管的活性;采用ELISA的方法测定小鼠免疫血清中HUVEC抗体水平;采用细胞毒T淋巴细胞(CTL)实验考察HUVEC-OK432疫苗诱导的细胞免疫应答水平。结果肺转移模型结果显示,在预防性和治疗性免疫中,HUVEC-OK432免疫均能有效降低小鼠肺上的肿瘤转移灶数目(P<0.01);皮内肿瘤血管模型的结果显示,HUVEC-OK432免疫能显著降低肿瘤新生血管的数目(116.5±20.6 vs 45.0±8.9,P<0.01);ELISA测定抗体的结果显示,HUVEC-OK432初次免疫后1周小鼠即产生了高滴度的特异性HUVEC抗体,随免疫次数的增加抗体水平不断升高,且该抗体在体外可以显著抑制HUVEC细胞的增殖;CTL实验的结果表明,HUVEC-OK432免疫组淋巴细胞体外有明显的特异性杀伤HUVEC的作用。结论 HUVEC-OK432疫苗免疫可以有效诱导小鼠产生靶向内皮细胞的免疫应答,从而有效抑制小鼠肝癌细胞转移。     

OK432优化的内皮细胞疫苗抗小鼠乳腺癌作用研究

目的探讨OK432优化的人脐静脉内皮细胞(HUVECs)疫苗对小鼠EAC乳腺癌的生长抑制作用。方法体外培养HUVECs,与佐剂OK432混合,制备HUVECs-OK432疫苗,以小鼠EAC乳腺癌皮下移植瘤模型考查HUVECsOK432疫苗的抗肿瘤效应,并通过ELISA、脾细胞增殖及细胞毒性T淋巴细胞(CTL)杀伤实验检测疫苗免疫后体液及细胞免疫应答水平。结果在预防性免疫中,HUVECsOK432疫苗可以明显抑制EAC乳腺癌生长;HUVECs-OK432疫苗诱导小鼠产生了高滴度的特异性HUVEC抗体;HUVECs-OK432疫苗能有效刺激免疫小鼠脾淋巴细胞的增殖;HUVECs-OK432组小鼠脾细胞诱导产生了明显的靶向HUVEC细胞的CTL杀伤作用。结论 HUVECs-OK432疫苗可以有效诱导机体产生靶向HUVEC的特异性体液及细胞免疫应答,从而有效抑制了小鼠EAC乳腺癌的生长。     

冻融抗原冲击致敏的树突状细胞抗肿瘤作用的研究

目的研究MC-38肿瘤冻融抗原致敏的树突状细胞(DC)对荷瘤小鼠是否有抗肿瘤作用。方法用MC-38肿瘤冻融抗原体外冲击致敏小鼠骨髓来源的DC,观察其诱导的CTL对MC-38肿瘤细胞的杀伤活性;体内以1×106DC/只多次接种于已荷瘤小鼠同侧腹股沟皮下,观察抗原冲击的DC对肿瘤生长的抑制作用以及对荷瘤小鼠生存期的影响。结果体外抗原冲击致敏的DC诱导的CTL对MC-38肿瘤细胞具有显著的杀伤作用,在效靶比为50∶1,25∶1,12.5∶1,6.25∶1时其杀伤率分别为68.84%,58.36%,41.56%,24.96%,;抗原冲击致敏的DC体内多次皮下免疫后对肿瘤的生长具有显著的抑制作用,能显著延长荷瘤小鼠的生存期。结论肿瘤冻融抗原致敏DC多次皮下免疫对荷瘤小鼠具有显著的抗肿瘤作用。     

猴头多糖抗肿瘤及对免疫功能的影响

目的　研究猴头多糖 ( HEPS)对小鼠 S1 80 肉瘤及免疫功能的影响。方法　 H EPS按 10 0、2 0 0、40 0m g/ kg体重连续灌胃 15 d,测定荷瘤小鼠瘤重 ,通过检测小鼠抗体生成细胞、迟发性变态反应、NK细胞活性 ,荷瘤小鼠免疫器官分别检测 HEPS对体液免疫、细胞免疫、非特异免疫及异常免疫的调节作用。结果　 HEPS可显著抑制 S1 80 肉瘤的生长 ,提高荷瘤小鼠胸腺和脾重 ,增强正常小鼠抗体形成细胞溶解绵羊红细胞能力、迟发型变态反应能力、NK细胞活性。结论　 HEPS具有抗肿瘤及免疫调节作用     

Dendritic cell-tumor coculturing vaccine can induce antitumor immunity through both NK and CTL interaction

Immunization of dendritic cells (DC) pulsed with tumor antigen can activate tumor-specific cytotoxic T lymphocytes (CTL) that are responsible for protection and regression. We show here that immunization with bone marrow-derived DC cocultured with tumor cells can induce a protective immunity against challenges to viable tumor cells. In this study, we further investigated the mechanism by which the antitumor activity was induced. Immunization of mice with DC cocultured with murine colon carcinoma. CT-26 cells, augmented CTL activity against the tumor cells. Concomitantly, an increase in natural killer (NK) cell activity was also detected in the same mice. When DC were fixed with paraformaldehyde prior to coculturing with tumor cells, most of the CTL and NK cell activity diminished, indicating that DC are involved in the process of presenting the tumor antigen(s) to CTL. NK cell depletion in vivo produced markedly low tumor-specific CTL activity responsible for tumor prevention. In addition, RT-PCR analysis confirmed the high expression of INF-gamma mRNA in splenocytes after vaccination with DC cocultured with tumors, but low expression in splenocytes from NK-depleted mice. Most importantly, the tumor protective effect rendered to DC by the coculturing with CT-26 cells was not observed in NK-depleted mice, which suggests that DC can induce an antitumor immune response by enhancing NK cell-dependent CTL activation. Collectively, our results indicate that NK cells are required during the priming of cytotoxic T-cell response by DC-based tumor vaccine and seem to delineate a mechanism by which DC vaccine can provide the desired immunity.     

Effective tumor immunity to melanoma mediated by B16F10 cancer stem cell vaccine

Although tumor vaccines have been considered a promising immunotherapy approach, therapeutic tumor vaccines are mostly disappointing in the clinic due to vaccine weak immunogenicity. Cancer stem cells (CSCs) may broaden the antigenic breadth and effectively induce the immune responses against autologous cancer cells. Here we report on the development of the B16F10 CD133⁺ CD44⁺ CSCs (B16F10 CSCs) vaccine to induce tumor immunity to melanoma in mice. Efficacy of against melanoma was evaluated by analysis of tumor growth and mouse survival. Immunogenicity was assessed by ELISA and flow cytometric assays, including serum cytokines, cytotoxic activity of NK cells and splenocytes in the immunized mice. The results showed that the B16F10 CSC vaccine resulted in tumor shrinkage and mouse lifespan extension. The cytotoxic activity and IFN-γ level were significantly increased in mice immunized with B16F10 CSC vaccine compared with the mice immunized with control vaccines. Additionally, New York esophageal squamous cell carcinoma-1, an efficient tumor associated antigen over-expressed by B16F10 CSCs, was markedly reduced in expression in melanoma tissue, suggesting decrease of CSC subpopulation due to B16F10 CSC vaccination. Collectively, the findings may represent a new powerful approach for treatment of melanoma by B16F10 CSC vaccination.     

Antitumor immunity of human SART3 gene vaccine against mouse tumor in vitro/vivo

To determine whether squamous cell carcinoma antigen recognized by human T cell 3 (SART3) gene can induce antitumor immunity against tumor cells which express the gene, we constructed mouse tumor cells expressing human SART3 (LM8-SART3) and carried out experiments in vitro/vivo. After subcutaneous injection with SART3 gene vaccine, cytotoxic T lymphocyte (CTL) activity in vitro was measured using Cell Counting Kit-8. As for the in vivo part, C3H mice were divided into several groups. One group was challenged with tumor cells after immunity. Another group was treated with the vaccine after tumor implantation. It was found that human SART3 DNA vaccine can elicit a specific CTL reaction from the mouse splenocytes. After vaccination, tumor occurrence and tumor growth speed was reduced. The vaccine also shows activity in tumor treatment. We conclude that the human SART3 DNA vaccine can induce antitumor ability against tumor cells expressing human SART3 (LM8-SART3) in vitro/vivo which may provide new possibilities in antitumor therapy.     

An aqueous extract of Limoniastrum guyonianum gall induces anti-tumor effects in melanoma-injected mice via modulation of the immune response

The objectives of this study were to evaluate the in vitro and in vivo anti-tumor potential of the aqueous gall extract (G extract) from Limoniastrum guyonianum and to elucidate its immunological mechanisms, in part, by assessing its effects on the growth of transplanted tumors and the immune response in these tumor-bearing mice. Here, mice were inoculated with B16F10 mouse tumor cells and then treated intraperitoneally with G extract at 25 or 50 mg extract/kg BW for 7, 14, or 21 days. At each timepoint, effects of the extract on the tumor growth, splenocytes proliferation, NK cell activity, and CTL activity among splenocytes isolated from the mice were measured. G extract-induced tumor growth inhibition was associated with characteristic apoptotic changes in the tumor cells, like nuclear condensation. In addition, the extract inhibited melanin synthesis and tyrosinase activity among melanoma cells in a concentration-related manner. G extract did not only significantly inhibit the growth of the transplantable tumor, but also remarkably increased splenocytes proliferation and both NK and CTL activities in tumor-bearing mice. The extract was also seen to have promoted lysosomal activity of host macrophages and gave rise to enhanced cellular anti-oxidant activity in several cell types in mice.     

Enhancement of antitumor effects of paclitaxel (taxol) in combination with red ginseng acidic polysaccharide (RGAP)

We have recently reported that red ginseng acidic polysaccharide (RGAP), isolated from Korean red ginseng (Panax ginseng C.A. Meyer), shows immunomodulatory and antitumor activities, mainly mediated by the nitric oxide (NO) production of macrophages. This compound may be used in cancer therapy alone or in combination with other chemotherapeutic agents. The synergistic effect of RGAP and paclitaxel (taxol) was evaluated to develop new biological response modifiers in cancer therapy. The present study demonstrates a synergistic antitumor effect of RGAP and paclitaxel in mice transplanted with sarcoma 180 and B16 melanoma. Combined treatment with paclitaxel (5 or 15 mg/kg) and RGAP (25 mg/kg) resulted in a 28.6 or 42.8 % increase in the life span of ICR mice bearing sarcoma 180 tumor cells, while no obvious effect was seen on sole paclitaxel treatment. When a combination of paclitaxel (10 mg/kg) and RGAP (100 mg/kg) was administered to C57BL/6 mice implanted with B16 melanoma, the tumor weight per mouse also decreased by 76.3 %, suggesting that RGAP may be used as an adjuvant in medicinal applications of paclitaxel. The augmented antitumor effect of paclitaxel is supposed to be the result of the immunomodulating antitumor effect of RGAP. RGAP, having B cell specific mitogenic activity, induced the secretion of interleukin-6 (IL-6) in spleen cells in a concentration-dependent manner (5 to 500 microg/microL). RGAP also restored the proliferation of splenocytes and NK cell activity suppressed by paclitaxel. Flow cytometric analysis of splenocytes in mice treated with paclitaxel showed a significant increase of CD11b+ cells. Additionally, a synergistic effect of RGAP and paclitaxel was found to effect an increased tumoricidal activity of macrophages. The above results suggest that clinical trials of RGAP as an adjuvant in cancer chemotherapy of paclitaxel are highly feasible.     

国产云芝多糖对小鼠抗肿瘤免疫反应的促进作用

本文采用观察同一个体多种指标的方法,研究国产云芝胞内多糖对C57BL/6J/J小鼠体内的黑色素瘤B16的抗瘤作用及其促进荷瘤鼠抗肿瘤免疫反应的作用.结果表明,这种云芝多糖对黑色素瘤B16的抑瘤率为40.8%,与环磷酰胺合用对黑色素瘤B16人工肺转移的抑制率达84.4%;其抗瘤作用机理以拮抗肿瘤的免疫抑制作用,多方面有效地促进行瘤鼠的非特异性抗肿瘤免疫反应为主,对正常机体的免疫反应也有一定促进作用.     

Protective antitumor activity through dendritic cell immunization is mediated by NK cell as well as CTL activation

Dendritic cells (DCs) are potent professional antigen-presenting cells (APC) capable of inducing the primary T cell response to antigen. Although tumor cells express target antigens, they are incapable of stimulating a tumor-specific immune response due to a defect in the costimulatory signal that is required for optimal activation of T cells. In this work, we describe a new approach using tumor-DC coculture to improve the antigen presenting capacity of tumor cells, which does not require a source of tumor-associated antigen. Immunization of a weakly immunogenic and progressive tumor cocultured with bone marrow-derived DCs generated an effective tumor vaccine. Immunization with the cocultured DCs was able to induce complete protective immunity against tumor challenges and was effective for the induction of tumor-specific CTL (cytotoxic T lymphocyte) activity. Furthermore, high NK cell activity was observed in mice in which tumors were rejected. In addition, immunization with tumor-pulsed DCs induced delayed tumor growth, but not tumor eradication in tumor-bearing mice. Our results demonstrate that coculture of DCs with tumors generated antitumor immunity due to the NK cell activation as well as tumor-specific T cell. This approach would be useful for designing tumor vaccines using DCs when the information about tumor antigens is limited.     

泥鳅多糖对小鼠脾细胞免疫应答的影响

目的:研究泥鳅多糖对小鼠脾细胞免疫应答的影响.方法:分别给正常小鼠、刀豆蛋白(ConA)或左旋咪唑免疫增强小鼠、环磷酰胺免疫抑制小鼠腹腔注射提取的泥鳅多糖,连续给药7d.用~H-胸苷掺入法测定T淋巴细胞的增殖.用~(51)Cr同位素标记法测定细胞毒T淋巴细胞的细胞毒性和天然杀伤细胞的活性.结果:泥鳅多糖5或10mg·kg~(-1)·d~(-1)可以提高T淋巴细胞的增殖,并增强细胞毒T淋巴细胞的细胞毒性和天然杀伤细胞的活性,还能增强ConA解除环磷酰胺对T淋巴细胞增殖抑制的能力,抑制率从环磷酰胺对照组的51.4％分别降低到18.2％和35.1％.而且,给予小鼠10或20mg·kg~(-1)·d~(-1)的泥鳅多糖,可以恢复环磷酰胺所致的天然杀伤细胞活性降低.结论:泥鳅多糖能增强小鼠脾细胞中T细胞、细胞毒T淋巴细胞和天然杀伤细胞的活性.     

Prevention of growth and metastasis of murine melanoma through enhanced natural-killer cytotoxicity by fatty acid-conjugate of protopanaxatriol

Ginsenosides, the glycosides of Panax ginseng, are metabolized (deglycosylated) by intestinal bacteria after oral administration. 20(S)-Protopanaxatriol (M4) is the main bacterial metabolite of protopanaxatriol-type ginsenosides and mediates their antitumor effects. To clarify the mechanism of the M4-mediated antitumor effect, the antitumor activity and metabolism of M4 was examined, using the C57BL/6 mice implanted with B16-BL6 melanoma. The chronic oral administration of M4 inhibited the growth of B16-BL6 melanoma at the implanted site. Analyses using TLC, HPLC, MS and NMR suggest that orally administered M4 was absorbed from the small intestine into the mesenteric lymphatics followed by the rapid esterification of M4 with fatty acids and its accumulation in the tissues including the liver and lung. The administration of M4 prior to the intravenous injection of B16-BL6 cells abrogated the enhanced lung metastasis in the mice pretreated with 2-chloroadenosine more effectively than in those pretreated with anti-asialo GM1. The esterified M4 (EM4) did not directly affect tumor growth in vitro, whereas it stimulated splenic NK cells to become cytotoxic to tumor cells. These results indicate that the antitumor activity of M4 is based on the NK cell-mediated tumor lysis enhanced by EM4.