针对HER-2的多肽疫苗CKL9与YL20的抗肿瘤活性研究

目的研究能够诱导激活细胞免疫应答的多肽疫苗CKL9、YL20在细胞水平和整体动物水平的抗HER-2阳性肿瘤作用,为肿瘤新药开发提供依据。方法采用CCK-8法检测CKL9、YL20诱导小鼠淋巴细胞的增殖作用,采用LDH法检测细胞毒性T淋巴细胞(CTL)的活性,采用动物体内实验评价CKL9、YL20的抗肿瘤活性。结果 CCK-8检测淋巴细胞增殖实验表明体外孵育多肽CKL9、YL20在一定程度上能够促进淋巴细胞增殖,在50 mg·L~(-1)孵育浓度下,CKL9、YL20相对增殖率分别为11.1%、16.7%;LDH法检测细胞毒性实验表明经多肽疫苗CKL9、YL20诱导分化的T淋巴细胞对HER-2阳性的肿瘤细胞有杀伤作用,当效靶比为80∶1时,细胞毒性T淋巴细胞对肿瘤细胞抑制率分别可达89.8%和84.3%;动物体内实验表明预免疫多肽CKL9、YL20能对BALC/c小鼠N87移植瘤产生明显抑制作用。结论基于HER-2的多肽疫苗CKL9、YL20具有免疫原性,可诱导特异性CD4和CD8 T淋巴细胞免疫,以抑制HER-2阳性肿瘤细胞生长。     

西洋参有效部位的抗肿瘤作用研究

目的　探讨西洋参有效部位 (Panaxquinquefolium'ef fectiveparts, PQEP)的抗肿瘤作用及其免疫调节作用。方法采用小鼠肝癌(H22)和胃癌(MFC)两种移植瘤的动物模型,以肿瘤抑制率、免疫器官重量、NK细胞活性和淋巴细胞增殖率作指标。结果　PQEP有明显的抗肿瘤活性和免疫调节作用。结论　其抗肿瘤作用和其免疫调节作用有明显的相关性,PQEP的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.     

蟾酥脂质体注射液的抗肿瘤作用

目的探讨蟾酥脂质体注射液(CS)的抗肿瘤作用。方法体内实验采用小鼠肝癌H22、肉瘤S180荷瘤小鼠模型,考察CS的抑瘤作用;采用小鼠接种肝癌H22后,用环磷酰胺造成免疫低下模型,考察CS提高机体免疫力作用。体外实验采用MTT法,测CS对人宫颈癌HeLa细胞或人肝癌BEL-7402细胞增殖抑制率。结果CS(0.2、0.4、0.8 mg.kg-1)对2种荷瘤小鼠肿瘤增长有显著或非常显著的抑制作用(P<0.05,P<0.01);同时对机体具有明显的免疫调节作用。CS 0.01~3.00 mg.L-1对人宫颈癌HeLa细胞或肝癌BEL-7402细胞增殖有非常显著的抑制作用;CS对人宫颈癌HeLa细胞IC50为0.51 mg.L-1,95%可信限0.213 2~1.214 0 mg.L-1,CS对人肝癌BEL-7402细胞IC50为0.75 mg.L-1,95%可信限0.312 6~1.801 5 mg.L-1。结论蟾酥脂质体注射液体内、体外均有抗肿瘤活性,且具有免疫调节功能。     

食管癌抗原和超抗原构建肿瘤疫苗的抗癌作用研究

目的利用食管癌肿瘤可溶性抗原(TSA)和超抗原(SEC)构建肿瘤疫苗,刺激外周血淋巴细胞,诱导产生细胞毒性T细胞(CTLs),对肿瘤细胞进行体内外杀伤作用研究,以探讨其抗肿瘤作用。方法外周血淋巴细胞经肿瘤疫苗作用,进行体外培养,诱导产生细胞毒性T细胞;细胞毒实验测定效应细胞杀伤活性;建立小鼠移植瘤模型,用肿瘤疫苗进行干预治疗,观察其治疗效果。结果经肿瘤疫苗刺激的淋巴细胞组诱导的CTLs对靶细胞杀伤活性显著高于单纯淋巴细胞组(P<0.05),对TSA来源的食管癌细胞具有选择性杀伤作用;体内研究发现肿瘤疫苗能显著减轻小鼠荷瘤负担,延长生存期。结论肿瘤可溶性抗原与超抗原SEC构建的肿瘤疫苗能产生高效特异性的抗肿瘤效果,显示出良好的抗肿瘤免疫治疗作用。     

五味子宁神口服液的免疫调节作用研究

目的:研究五味子宁神口服液(WOL)的免疫调节作用.方法:采用测定小鼠碳粒廓清率、二硝基氟苯(DNFB)诱导小鼠迟发型超敏反应、血清溶血素抗体IgM水平,以及测定ConA和LPS诱导的脾T、B淋巴细胞的增殖转化实验(MTT法).结果:WOL提高小鼠血清溶血素抗体IgM水平及免疫功能低下小鼠的吞噬指数、吞噬系数、耳肿胀度、脾和胸腺指数,增强ConA和LPS诱导的脾T、B淋巴细胞增殖转化反应.结论:WOL对小鼠具有一定的免疫增强作用.     

拳参乙醇提取物的免疫调节作用

目的 研究拳参乙醇提取物(BRE)的免疫调节作用.方法 在小鼠体外实验中,通过比色分析法检测小鼠巨噬细胞系RAW264.7细胞吞噬中性红的能力.在小鼠体内实验中,通过测定胸腺、脾脏重量并计算脏器指数;MTT法检测T淋巴细胞增殖和NK细胞活性;鸡红细胞免疫后测定小鼠血清溶血素抗体水平;ELISA法测定血清中IL-2含量....     

板栗中蛋白质的分离鉴定及活性研究

目的探讨板栗中蛋白质的分离纯化及其促肾细胞生长作用和抗肿瘤作用及机制。方法从板栗中提取出的水溶性蛋白质,经鉴定是欧栗球蛋白(CAS)。利用MTT法检测该蛋白对正常肾细胞和肝癌细胞HepS,小鼠肉瘤S180腹水型肿瘤细胞增殖的影响;检验其对小鼠T细胞和B细胞增殖作用的影响;用DNA电泳方法检测该蛋白诱导HepS细胞凋亡的作用。结果 CAS在体外对正常肾细胞的增殖有促进作用,对试验的肿瘤细胞的增殖均有抑制作用,对T细胞和B细胞的增殖有促进作用,对HepS细胞具有诱导凋亡的作用。结论 CAS具有促肾细胞生长和抗肿瘤作用,其抗肿瘤作用机制可能通过提高机体免疫力和诱导细胞凋亡来实现。     

灵龙颗粒剂的抗肿瘤作用及对免疫功能的影响

目的观察灵龙颗粒剂(简称LLE)抗动物移植性肿瘤和裸鼠人肺癌与体外抑瘤作用及其对荷瘤小鼠免疫功能的影响.方法体内抗肿瘤实验采用腋下接种肿瘤细胞及实体瘤称重的方法;腹腔接种瘤液观察荷瘤动物存活率;裸鼠腋下接种人肺腺癌及实体瘤称重;体外抗肿瘤实验采用直接观察肿瘤细胞增殖的MTT法;2,4-二硝基氟苯诱发的DTH反应采用小鼠耳片差重法;淋巴细胞转化实验采用MTT法检测;TNF-α活性检测采用L929细胞杀伤实验.结果LLE(13.1,17.5,35.0g生药*kg-1)对移植性动物肿瘤S180及H22的抑瘤率分别为47.7%,66.1%,82.0%及67.0%,73.3%,76.4%;使荷EAC肿瘤小鼠的生存期延长; LLE(17.5,35.0g生药*kg-1)对裸鼠人肺腺癌有较强的抑制作用,抑瘤率为60.5%和77.8%;该药还显示一定的体外抑瘤作用;LLE对荷S180及H22小鼠的DTH反应有增强作用;对淋巴细胞转化有明显促进作用;LLE可有效增加荷瘤小鼠腹腔巨噬细胞TNF-α的产生.结论ig LLE对荷瘤小鼠具有显著的抗肿瘤作用,且能拮抗荷瘤所致的免疫功能的抑制.     

当归多糖组分AP-3对不同淋巴细胞亚群的作用

目的观察当归多糖组分AP-3对不同淋巴细胞亚群的作用。方法免疫磁珠法制备T、B细胞亚群;四甲基偶氮唑蓝(MTT)法检测细胞增殖;流式细胞术观察CD4+T细胞亚群比例的变化。结果在30~300μg/ml范围内,AP-3能显著促进小鼠脾淋巴细胞、巨噬细胞、混合淋巴细胞和T淋巴细胞的增殖反应,而对B细胞增殖有一定的抑制作用;显著增加培养的脾细胞中CD4+T细胞亚群的比例。结论当归多糖具有独特的免疫调节特性,活化巨噬细胞、T细胞以及辅助性T细胞(Th细胞),但对B细胞有一定的抑制作用。     

In vitro anti-inflammatory effects of cynaropicrin, a sesquiterpene lactone, from Saussurea lappa

We investigated in vitro anti-inflammatory effects of cynaropicrin, a sesquiterpene lactone from Saussurea lappa, on tumor necrosis factor (TNF)-alpha and nitric oxide (NO) release, and lymphocyte proliferation. Cynaropicrin strongly inhibited TNF-alpha release from lipopolysaccharide-stimulated murine macrophage, RAW264.7 cells, and differentiated human macrophage, U937 cells, proved to produce notable amount of TNF-alpha. It also potently attenuated the accumulation of NO released from lipopolysaccharide- and interferon-gamma-stimulated RAW264.7 cells in a dose-dependent manner. In addition, the immunosuppressive effects of the compound on lymphocyte proliferation in response to mitogenic stimuli were examined. Cynaropicrin also dose-dependently suppressed the proliferation of lymphocytes from splenocytes and interleukin-2-sensitive cytotoxic T lymphocyte, CTLL-2 cells, stimulated by lipopolysaccharide, concanavalin A, phytohemagglutinin and interleukin-2. However, treatment with sulphydryl compound, L-cysteine, abrogated all these inhibitory effects. These results suggest that cynaropicrin may participate in the inflammatory response by inhibiting the production of inflammatory mediators and the proliferation of lymphocytes and its inhibitory effect is mediated through conjugation with sulphydryl groups of target protein(s).     

浒苔多糖的分离纯化及其对小鼠淋巴细胞活性的影响

为了探讨浒苔(Enteromorpha)多糖的免疫调节活性,采用Sevage法,Sephacryl-300 分子筛层析以及乙醇分级沉淀法获得四个浒苔多糖组分EP,EP1、EP2和EP3,比较四种多糖对体外培养的小鼠T、B淋巴细胞增殖影响;同时观察作用效果较强的EP2在T、B淋巴细胞中的分布及其对细胞RNA合成的影响。结果表明,EP、EP1、EP2和EP3中单糖、糖醛酸、硫酸根含量存在差异,均能促进对B淋巴细胞以及ConA或LPS诱导的T、B淋巴细胞增殖,且EP2的作用效果最好;经EP2处理的T、B淋巴细胞或同时用EP2和丝裂原(ConA或LPS)处理的T、B淋巴细胞体积明显增大,RNA合成增加,B淋巴细胞有母细胞化趋势。由此可见,浒苔多糖能调节小鼠淋巴细胞活性,促进丝裂原诱导的淋巴细胞分化和免疫应答,有望成为一种新的动物免疫增强剂。     

黄芪皂苷Ⅳ对小鼠T、B淋巴细胞增殖和腹腔巨噬细胞功能的影响

目的：探讨黄芪皂苷Ⅳ（ASI）对小鼠T,B淋巴细胞增殖和腹腔巨噬细胞分泌的细胞因子的影响。方法：采用MTT法检测T,B淋巴细胞的增殖;采用分光光度计测定法检测抗体活性;IL－1活性用胸腺细胞增殖法测定;TNF－α活性用L9292细胞杀伤法测定。结果：1）ASI50－200mg/kg ig7天能够促进T淋巴细胞增殖和抗体生成,而ASI50－100mg/kg 能够促进B淋巴细胞增殖,但是200mg/kg对B淋巴细胞增殖无影响;（2）ASI体外仅在200nmol/L对T,B淋巴细胞有促进作用;（3）ASI 1nmol/L可以促进腹腔巨噬细胞分泌IL－1,而100－1000nmol/L则抑制腹腔巨噬细胞分泌IL－1;（4）ASI体外可以抑制LPS刺激或无LPS刺激下的腹腔巨噬细胞分泌TNF－α。结论：ASI能够促进小鼠T,B淋巴细胞的增殖和抗体生成,同时可以抑制腹腔巨噬细胞体内分泌IL－1和TNF－α。     

Cellular and humoral systemic and mucosal immune responses stimulated in volunteers by an oral polybacterial immunomodulator "Dentavax"

The oral polybacterial immunomodulator Dentavax (D), composed of killed cells from Klebsiella pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, Candida albicans and Lactobacillus acidophilus and their lysates was created for immunoprophylaxis and therapy of oral mucosa and parodont inflammations. The stimulating effect of the preparation was evaluated in twelve volunteers immunized for 10 consecutive days. On days 7, 14, 21, 28 and 49 after the last immunization peripheral blood (PB) lymphocyte subsets, T lymphocyte activation and PB phagocytic activity, were studied by flow cytometry. PB lymphocyte proliferative responses to PHA, rIL-2, LPS and D were evaluated radiometrically. The production of TNF-alpha in supernatants of in vitro stimulated lymphocytes and specific IgA, IgM and IgG antibodies in serum and saliva was determined by ELISA. Ultrastructural morphologic changes in T and B lymphocyte populations were also investigated. Although no significant changes in the levels of basic lymphocyte subsets were detected, the early/late (CD57+/CD57-) CD8 T effectors ratio was increased at the end of the studied period, as were the percentage of PHA-responding (CD69+) T cells and PB phagocytizing cells. The most prominent lymphoprolipherative responses were measured upon costimulation with LPS+D and PHA+D on day 21. Electron-microscopic studies demonstrated a significant effect of D on both T and B cell activity. TNF-alpha concentration increased progressively from day 7 till the end of the investigation. Maximal concentrations were observed after stimulation with D and LPS. An increased level of specific salivary and serum antibodies against the components of D was found, with highest levels between days 7 and 21. Specific secretory IgA predominated in saliva as compared to IgM and IgG. Our results demonstrate the stimulating effect of Dentavax on PB lymphocyte functional activity and the specific humoral systemic and mucosal immunity.     

蝙蝠娥被孢霉多糖抗肿瘤和免疫功能研究

从蝙蝠娥被孢霉液体发酵的菌丝体中,采用热水提取,65%乙醇沉淀,DEAE-52和G-100分子筛分离、纯化得到了一个组分均一的天然多糖—蝙蝠娥被孢霉多糖(CPSM,Cordyceps Polysaccharides of Mortierella hepiali),其由甘露糖、葡萄糖和半乳糖3种单糖组成。体内实验表明,该多糖对小鼠移植瘤有明显抑制生长作用,但是在体外实验中对人肺癌细胞株A549、人肝癌细胞株Hep-G2、急性粒细胞白血病细胞株HL-60均没有表现出细胞毒作用。推测该CPSM是通过调节小鼠免疫系统的功能达到抑制移植瘤生长作用的。对移植瘤小鼠进行了T、B淋巴细胞增殖和NK细胞活性的实验表明灌胃给与多糖的小鼠组T、B淋巴细胞的增殖明显快于正常组,NK细胞活性大于正常组。而CPSM在体外对于小鼠腹腔巨噬细胞同样具有活化功能,增强巨噬细胞NO的分泌。     

Decreased lymphocyte blastogenesis,IL2 production and NK activity following nifedipine administration to healthy humans

Summary The effects of a single oral dose of nifedipine on part of the immune response in healthy humans has been investigated in terms of two different immune functions: T lymphocyte proliferation and NK activity. Both functions are known to require calcium ions.Ten healthy subjects were bled before and 30 min, and 4 and 24 h after receiving 10 mg nifedipine. Lymphocyte proliferation, both in mitogen-activated lymphocyte cultures, and in autologous and allogeneic mixed lymphocyte reactions, was significantly reduced (up to 48%) 30 min after drug administration and reverted to normal 4 h later. The inhibition could be attributed to reduction in IL2 production by the T cells isolated 30 min following the administration of nifedipine, since they normally express IL2-receptors. The addition of recombinant IL2 of 200 U·ml^–1 to the cell cultures restored their responsiveness.NK activity was significantly reduced 30 min and 4 h after drug administration and returned to normal at the 24th h. This function was also restored by the addition of IL2.The data suggest that calcium channel blockers may inhibit, at least transiently, lymphocyte functions in vivo.List of abbreviations AMLC Autologous mixed lymphocyte cultures - CD Cluster determinants - ConA Concanavallin A - CTL Cytotoxic T lymphocytes - GAM-FITC Fluorescein isothiocyanate-conjugated goat anti-mouse immunoglobulins - HLA-class II DP-DQ-DR antigens - IL2 Interleukin 2 - mAb Monoclonal antibodies - MHC Major histocompatibility gene complex - MLC Mixed lymphocyte cultures - NK Natural killer activity - non- T/T type AMLC Autologous mixed lymphocyte culture in which non-T cells are used as stimulators - PBL Peripheral blood lymphocytes - T/T type AMLC Autologous mixed lymphocyte culture in which PHA-activated T lymphocytes are used as stimulators     

Immunomodulatory activity of polysaccharides isolated from Strongylocentrotus nudus eggs

Our previous work showed that SEP, a novel glucan isolated from the eggs of sea urchins, Strongylocentrotus nudus, had remarkable anti-tumor activity. To elucidate the mechanism of the anti-tumor activity, the immunomodulatory activity of SEP was investigated. The in vivo experiment results showed that SEP remarkably enhanced spleen and thymus index in S180-bearing mice, and also stimulated ConA-induced splenocyte proliferation. Immunomodulatory activity assay in vitro indicated SEP could significantly enhance the mouse splenocyte proliferation in a dose-dependent manner. According to comitogenic activity tests, SEP showed significant comitogenic activities and adjuvant properties. We also demonstrated that SEP had a unique mode of immunostimulation with regard to its cell-type specificity. In other words, SEP markedly stimulated B and T cell proliferation, however the influence on B cells was greatly weaker than that on T cells. IL-2, TNF-alpha, and IFN-gamma mRNA expression was upregulated after the mouse splenocytes were treated by SEP, indicating that Th1 cell was the primary cellular target affected by SEP on T lymphocyte. SEP enhanced production of nitric oxide (NO), upregulated mRNA expression of inducible nitric oxide synthase (iNOS) in peritoneal macrophages in a dose-dependent manner. In addition, SEP did not show direct toxicity to tumor cells. Consequently, the anti-tumor effect of SEP was related to stimulating host immunity/enhancing the immune system functions, which may mainly result from SEP activating lymphocytes and macrophages and stimulating secretion of some cytokines.     

乙双吗啉对有丝分裂原诱导的小鼠淋巴细胞增殖反应的抑制作用

当浓度为1.56mg/L时,乙双吗啉就可明显抑制LPS诱导的体外小鼠淋巴细胞增殖反应,其作用随浓度的增高而加强。当此药浓度增大到6.25mg/L时,ConA和PHA诱导的体外小鼠淋巴细胞增殖反应才开始出现明显抑制。表明乙双吗啉对B细胞增殖反应的抑制作用强于对T细胞增殖反应。乙双吗啉对淋巴细胞无非特异细胞毒作用。     

Immunomodulatory effect of caffeic acid phenethyl ester in Balb/c mice

Caffeic acid phenethyl ester (CAPE), an the active component of propolis, is known to have anticarcinogenic, antiviral and various biological activities; however, the effect of CAPE on the immunomodulatory activity in vivo remains unknown. We have investigated the effect of CAPE on the immune system in female Balb/c mice. CAPE (0, 5, 10, 20 mg/kg) was given to mice orally for 14 days. Immunomodulatory activity was evaluated by assessment of body and organ weight, lymphocyte blastogenesis, plaque-forming cell (PFC) assay, lymphocyte subpopulation by flow cytometry and cytokine production. Even though the change of body weight was not observed in CAPE-administered group, thymus weight and/or cellularity of thymus and spleen are decreased at the all dose groups of CAPE (5, 10, 20 mg/kg). On the other hand, CAPE had no effect on B lymphocyte proliferation induced by lipopolysaccharide (LPS) but increased T lymphocyte blastogenesis induced by concanavalin A (Con A) at the dose of 20 mg/kg. In the case of lymphocyte subpopulation, the population of T and B cells was not changed but CD4(+) T cell subsets are significantly increased in exposure to CAPE. The antibody responses to T lymphocyte dependent antigen, sheep red blood cell and keyhole limpet hemocyanin (KLH) were increased more than 10 mg/kg in CAPE-treated group. Likewise, the cytokine, IL-2, IL-4 and IFN-gamma were significantly increased at the dose of 20 mg/kg CAPE group. These results suggest that CAPE could have immunomodulatory effects in vivo.     

白芷提取物对大鼠脾淋巴细胞免疫抑制作用初探

目的探讨白芷提取物对地塞米松致大鼠脾淋巴细胞免疫抑制作用的影响。方法建立地塞米松致大鼠免疫抑制模型,考察白芷提取物对大鼠脾淋巴细胞的增殖活性、自然杀伤细胞（NK细胞）活性的影响。结果白芷提取物（5．0g/kg）对刀豆蛋白A（ConA）诱导脾淋巴T细胞增殖能力有升高作用,但对脂多糖（LPS）诱导脾淋巴B细胞增殖能力未见明显影响,对脾淋巴细胞的NK细胞活性也有增强作用。结论白芷提取物（5．0g／kg）曲能提高大鼠脾淋巴T细胞和NK细胞的增殖活性。