小鼠肝癌细胞(H22)源外泌体的体内抗肝癌作用研究

目的在荷瘤动物模型观察小鼠肝癌细胞（H22）来源的外泌体（exosomes）激发宿主抗肝癌免疫应答的效应。方法用自行分离纯化的小鼠肝癌细胞H22源外泌体免疫小鼠,然后给小鼠皮下注射H22肿瘤细胞,观察肿瘤的生长状况。用MTT法检测小鼠脾细胞增殖情况和细胞毒活性;用免疫组织化学法检测肿瘤组织中CD4^＋、CD8^＋T淋巴细胞浸润情况。结果该外泌体使肿瘤出现时间延迟,肿瘤生长缓慢,小鼠生存率提高。促进脾淋巴细胞增殖并增强其细胞毒活性,促进CD4^＋和CD^8＋T细胞活化。结论小鼠肝癌细胞（H22）源外泌体能诱导抗肿瘤应答,有效地诱导特异性的杀伤肿瘤细胞活性,有针对亲本细胞的免疫保护作用。     

小鼠免疫力对接种人类肝癌细胞系9724生长的影响

目的 探讨不同免疫力小鼠的免疫细胞对人类肝癌细胞系9724生长的影响及异种特异性细胞毒杀伤能力。方法 将培养后的人类肝癌细胞系接种到不同免疫力小鼠（免疫正常的BALB／c,B缺陷的CBA／N,T缺陷的BALB/c nude及T,B缺陷的C.B－17 SICD小鼠）右后臀皮下,观察其生长;取实验和相应的正常鼠脾细胞进行细胞毒杀伤试验。结果 BALB/c和CAB／N小鼠不成瘤;BALB/c nude和C.B－17 SCID小鼠100％成瘤;SCID免疫重建组用BALB/c外周淋巴细胞重建SCID（BALB/c－PBL－SCID）的不成瘤,用CAB／N外周淋巴细胞重建的SCID（CBA／N－PBL－SCID）成瘤,接种过瘤的BALB/c和CBA／N小鼠脾细胞对癌细胞有较强的细胞毒杀作用,对照组和nude,SCID小鼠无杀伤作用;SCID免疫重建有较小的杀伤作用。结论 T细胞在异种瘤移植排斥中起主要的免疫杀伤作用,这种排斥是异种特异笥的;肿瘤免疫需要综合性免疫作用。     

人多发性骨髓瘤荷瘤SCID小鼠模型的建立及其生长特性

为研究人多发性骨髓瘤(Multiple Myeloma，MM)在SCID小鼠免疫重建前后体内生长的特性和免疫细胞对肿瘤的作用，建立人多发性骨髓瘤(MM)荷瘤SCID小鼠模型。体外培养IL-6依赖的人MM细胞株XG7，接种于SCID小鼠和用人淋巴细胞重建的SCID小鼠皮下；监测成瘤状态，通过免疫组化观察细胞毒性T淋巴细胞(Cytotoxic T lymphocytes，CTLs)杀伤。结果表明：成瘤后的肿瘤细胞CD28、CD138等分子的表达与接种前没有明显变化；免疫重建后的SCID小鼠成瘤潜伏期延长，瘤体积小；免疫重建后的SCID小鼠肿瘤组织中，有人CD3 T细胞的浸润。结果提示：SCID小鼠是建立肿瘤模型和肿瘤免疫研究的良好动物；免疫重建后的SCID小鼠体内肿瘤生长受抑制：人MM细胞在小鼠体内的生长状态与淋巴细胞免疫有关。     

新型宫颈癌治疗性疫苗肿瘤杀伤作用

目的研究新型治疗性疫苗NTV的体内外宫颈癌杀伤作用。方法采用51Cr释放法研究脾淋巴细胞对宫颈癌细胞TC-1的体外杀伤活性,将正常小鼠体内激活的免疫活性细胞输给荷瘤小鼠,采用过继性细胞毒性T淋巴细胞(CTL)转移实验研究CTL的体内抗肿瘤作用。结果 CTL杀伤实验设置的效应细胞与靶细胞数量比分别为3:1、10:1及30:1,51Cr释放分析结果显示,疫苗NTV能够显著诱导脾淋巴细胞的肿瘤特异性杀伤作用,杀伤作用呈剂量依赖性变化,杀伤效率最高可达到53.5%;疫苗NTV在过继性CTL转移实验中的肿瘤抑制率为64.7%,上述结果均与阴性对照组有统计学差异(P<0.05)。结论疫苗NTV在体内外对宫颈癌细胞均具有良好的杀伤作用。     

肝癌细胞疫苗对荷瘤小鼠脾淋巴细胞IFN-γ生成及肿瘤浸润淋巴细胞的影响

目的探讨高强度聚焦超声(HIFU)制备肿瘤疫苗对小鼠抗同源肿瘤免疫力的影响。方法以HIFU辐照或高温水浴灭活H22细胞制成HIFU瘤苗和高温瘤苗。将雌性BALB/c小鼠108只,随机分为HIFU瘤苗、高温瘤苗、生理盐水组,分别皮下注射HIFU瘤苗、高温瘤苗、生理盐水,之后接种H22细胞,观测荷瘤小鼠肿瘤体积及生存期。ELSIA检测免疫后荷瘤小鼠脾淋巴细胞在同源肿瘤再刺激下IFN-γ的生成。免疫组化检测小鼠肿瘤浸润T淋巴细胞。结果 HIFU瘤苗免疫使小鼠肿瘤生长被抑制,脾淋巴细胞IFN-γ生成增加,且较高温瘤苗组更显著(P<0.05)。瘤苗增加了肿瘤中CD4+和CD8+细胞浸润的程度。结论 HIFU瘤苗提高了实验动物的抗肿瘤免疫力,表现为增高的淋巴细胞IFN-γ生成和肿瘤内淋巴细胞浸润。     

大蒜素诱导鼠肝癌细胞MM45T.Li 细胞株凋亡

目的探讨大蒜素对体外培养的鼠肝癌细胞MM45T.Li生长的抑制和诱导细胞凋亡的作用. 方法用MTT法检测药物效应,透射电镜及流式细胞仪观察大蒜素处理MM45T.Li细胞后细胞凋亡和细胞周期的改变.结果① MTT法证实大蒜素对MM45T.Li细胞生长有抑制作用,呈浓度、剂量-效应关系.② DAN琼脂糖凝胶电泳可见细胞凋亡特征性"DAN梯形".③流式细胞仪检测发现大蒜素处理MM45T.Li细胞后细胞周期滞留于G0/G1期,对照组及实验组细胞凋亡率分别为(4.98±4.841)%、(20.91±5.272)%,并可见凋亡峰.④透射电镜可见MM45T.Li细胞出现凋亡细胞形态学改变.结论大蒜素对鼠肝癌细胞MM45T.Li有抑制作用,其抑制作用可能与诱导肝癌细胞凋亡、阻抑细胞周期有关.     

树突状细胞疫苗联合抗PD-L1抗体治疗小鼠肝癌的实验研究

[目的]探讨抗PD-L1抗体能否增强树突状细胞疫苗对小鼠肝癌的治疗效果。[方法]将H22肝癌细胞接种于C57BL/6J小鼠左侧腋窝皮下。5 d后,小鼠被随机分为4组,分别给予树突状细胞疫苗治疗、抗PD-L1抗体治疗,树突状细胞疫苗联合抗PD-L1抗体治疗,以及未治疗组。肿瘤接种后28 d,处死小鼠,提取各组小鼠脾淋巴细胞并分析其活性。通过测量肿瘤大小来判定抗肿瘤效应。[结果]与未治疗组相比,各治疗组T细胞活性显著增加,肿瘤生长明显受抑制。[结论]抗PD-L1抗体能够增强树突状细胞疫苗治疗小鼠肝癌的效果,可能于T细胞的活性增强有关。     

负载肿瘤抗原的树突状细胞疫苗诱导CTL的抗肿瘤效应

目的:探讨负载肿瘤抗原的树突状细胞(DC)疫苗诱导的细胞毒性T淋巴细胞(CTL)的体外杀瘤作用及对荷瘤小鼠的治疗作用.方法:体外培养的小鼠骨髓树突状细胞用小鼠结肠腺癌细胞株CT26细胞抗原冲击致敏制备负载肿瘤抗原的DC疫苗;负载肿瘤抗原的DC疫苗激活同源T淋巴细胞产生细胞毒性T淋巴细胞(CTL),采用乳酸脱氢酶(LDH) 4 h释放法检测CTL在体外对CT26细胞的杀伤活性;建立CT26荷瘤小鼠模型,应用CTL治疗荷瘤小鼠,观察肿瘤大小和小鼠存活期.结果:负载CT26肿瘤抗原的DC疫苗能够诱导T细胞增殖分化为肿瘤特异CTL,该CTL对CT26细胞有高效而强烈的杀伤作用,杀伤率为(83.95±11.25)%,而对B16细胞、3LL Lewis 细胞无明显杀伤作用,杀伤率分别为(12.75±5.36)%和(11.38±4.57)%.应用CTL治疗荷瘤小鼠能显著抑制荷瘤小鼠肿瘤的生长,延长荷瘤小鼠存活期.结论:负载肿瘤抗原的DC疫苗能够诱导高效而特异的CTL杀瘤活性并能治疗荷瘤小鼠.提示负载肿瘤抗原的DC疫苗诱导的CTL可能在肿瘤的免疫治疗中发挥重要作用.     

携带flk-l的减毒沙门疫苗菌抗胶质瘤血管生成的研究

目的观察表达鼠血管内皮生长因子受体2(VEGFR2,flk-1)的重组减毒鼠伤寒沙门疫苗菌对胶质瘤荷瘤小鼠的抗肿瘤血管及肿瘤生长抑制作用。方法构建真核表达载体pcDNA3 1.-flk-l,通过电转化法将pcDNA3 1.-flk-1导入减毒鼠伤寒沙门菌SL7207中,经由胃管饲于C57BL/6J小鼠,对胶质瘤荷瘤小鼠进行免疫预防及治疗。通过观察免疫动物的生存期,免疫荧光法检测肿瘤血管密度,评价重组疫苗菌的抗血管及肿瘤生长抑制作用。分离免疫小鼠的脾细胞,分析重组疫苗菌免疫后小鼠体内的特异性细胞毒性T细胞(CTL)应答。结果重组疫苗菌免疫能够明显减少肿瘤血管密度,延缓胶质瘤的生长,延长小鼠生存期,获得明显的抗肿瘤效果。重组疫苗菌免疫后可诱导小鼠脾淋巴细胞产生针对flk-l的CTL活性。结论表达鼠flk-l的重组减毒鼠伤寒沙门疫苗菌经口服免疫,可打破小鼠对于自身抗原flk-1的免疫耐受,诱导小鼠产生抗flk-1的特异性免疫反应,特异性杀伤肿瘤血管内皮细胞,预防和治疗小鼠胶质瘤。     

树突状细胞疫苗联合CpG-ODN对膀胱肿瘤细胞的免疫学效应研究

目的 探讨负载肿瘤抗原的树突状细胞(DC)疫苗联合含未甲基化胞嘧啶鸟嘌呤(CpG)序列的寡脱氧核糖核苷酸(CpG-ODN)对膀胱肿瘤细胞的免疫学效应.方法 用小鼠BTT739细胞反复冻融抗原致敏培养第7天的DC,48h后分别加入CpG-ODN及肿瘤坏死因子-α(TNF-α)诱导DC成熟,制备负载肿瘤抗原的DC疫苗,灭活后用于刺激T淋巴细胞制备效应细胞(CTL),乳酸脱氢酶释放实验测定各组CTL对BTT739细胞的杀伤活性,ELISA法测定效应细胞培养上清液的干扰素-γ(IFN-γ)水平,同时检测了CTL对自体淋巴细胞的杀伤活性.结果负载肿瘤抗原的DC诱导的效应细胞分泌较高水平的IFN-γ,并对BTT739细胞具有较高的杀伤效应,CpG-ODN活化的DC疫苗诱导的效应细胞分泌IFN-γ水平及对肿瘤细胞的杀伤活性均高于TNF-α活化的DC疫苗诱导的效应细胞(P<0.01).DC疫苗对自体淋巴细胞无明显免疫杀伤活性.结论 CpG-ODN可增强负载肿瘤抗原的DC疫苗对膀胱肿瘤细胞的免疫杀伤效应,并且对自体淋巴细胞无免疫杀伤活性,为膀胱癌的免疫治疗提供了实验基础.     

MIF基因修饰的瘤苗免疫小鼠后T细胞数量和功能的分析

目的：研究了细胞在巨噬细胞游走抑制因子（MIF）基因修饰的瘤苗诱导抗肿瘤免疫反应中的应用。方法：将重组MIF腺病毒转染小鼠FBL3红白血病细胞制成的瘤苗接种于小鼠体内,观察脾脏和淋巴结中T细胞数量和功能的改变。结果：经MIF基因修饰的瘤苗免疫后,小鼠脾细胞和腹股沟引流淋巴结细胞数量明显增多,脾细胞CTL活性显增强,引流淋巴结中CD3^ 、CD28^ 、CD4^ 和CD8^ T细胞百分率均较对照组增加。MIF基因修饰的瘤苗免疫对小鼠受野生型FBL3肿瘤细胞再攻击具有明显的保护作用。结论：MIF基因修饰的瘤苗能诱导T细胞介导的抗肿瘤免疫反应,有可能作为肿瘤基因治疗的候选。     

融合基因GM-CSF/IL-2转基因瘤苗在肝癌特异性免疫治疗中的应用

目的 制备人白细胞介素2(hIL-2)与鼠粒-单核细胞集落刺激因子(mGM-CSF)融合基因修饰的H22肝癌全细胞瘤苗,探索融合基因GM.CSF/IL-2转基因瘤苗,在肝癌主动免疫治疗中特异性抗肿瘤作用.方法 用含hlL-2与mGM-CSF融合基因的真核表达载体在体外转染H22肝癌细胞,制成疫苗,皮下接种Balb/c小鼠,同时建立Balb/c小鼠荷瘤模型,ELISA法检测H22/pcDNA3.1( ).GM-CSF/IL-2瘤苗免疫组小鼠与各对照组小鼠(分别接种H22/pcDNA3.1( )瘤苗、H22瘤苗、PBS)血清中IL-10、IFN- γ水平,观察小鼠存活期;同时用51Cγ释放法测定H22/pcDNA3.1( )-GM-CSF/IL-2瘤苗免疫组小鼠与各对照组小鼠(单纯荷瘤组、正常组)脾细胞对亲本H22肝癌细胞的杀伤活性.结果 成功制备了含有hIL2与mGM-CSF融合基因的H22肝癌瘤苗,转基因瘤苗免疫组小鼠的脾细胞在体外对亲本H22肝癌细胞的杀伤率为38.3%,显著高于对照组小鼠的脾细胞对亲本H22肝癌细胞的杀伤率(分别为13.6%和7.5%),也高于其对S180细胞的杀伤率(9.1%)(P＜0.05).转基因瘤苗免疫组小鼠血清IFN-γ较对照组明显升高(P＜0.01),血清IL-10较对照组明显降低(P＜0.01).同时,转基因瘤苗免疫组小鼠生存期亦有明显延长.结论 转染hIL-2与mGM-CSF融合基因的同系肿瘤细胞瘤苗可激发特异性细胞介导的免疫反应,改善抗肿瘤免疫反应,延长荷瘤小鼠生存期.     

Therapeutic antitumor response to cervical cancer in mice immunized with U14 v accines transfected with costimulatory B7 gene

Objective To investigate the effect of U14 vaccine transfected with the B7 gene in inducin g antitumor immune response to murine cervical carcinoma in Chinese 615-strain mice.Methods A recombinant retroviral plasmid vector expressing mouse B7-1 gene (pLNSX-mB7) was transfected into 615-strain mouse cervical carcinoma cell line No. 14 (U1 4) by electroporation to set up a highly-expressed mB7-1 U14 cell clonal strai n (B7(+)U14). In vivo experiments: (1) B7(+)U14 vaccine was primed to protect t he 615-strain mice against U14 re-challenge. (2) B7(+)U14 vaccine was injecte d into tumor-bearing mice with different tumor sizes. Lifetimes and tumor s izes were recorded. In vitro cytotoxicity assay: Mice were immunized with B 7(+)U14 or U14 vaccine and 2 weeks later, spleen cells of those mice were cultur ed for 2 days. The cytotoxicity of these cells against U14 was detected by 5-d iphenyl tetrazolium bromide assay.Results We obtained several B7-1 high expression clonal U14 lines. In vivo experiment, we did not find tumor growing in 3 of the 6 mice primed by B7(+)U14 vaccine during their entire life after re-challenge with U14. The other 3 mice develo ped tumors and their average survival time was longer than that of the control g roup (P&lt;0.01). All 6 mice grew tumors in the control group. When the transplanted tumors became palpable, the mice were randomly divided into 3 group s to be injected with B7(+)U14 vaccine. It was effective for tumor-bearing mic e only when the tumor diameters were &lt;3 mm. When the diameters were ≥3 mm, it was not efficacious to inject B7(+)U14 vaccine (P&lt;0.05). In vitro cytotoxicity assay, cytotoxic T lymphocytes induced by B7(+)U14 vaccine h ad a high er cytotoxicity against U14 than that induced by U14 vaccine (F=310.8, P &lt;0.001).Conclusions Vaccines of cervical cancer cells transfected with the costimulatory molecule B7 gene can induce antitumor immune protection in host mice against U14 re-challe nge. This treatment may cure part of the tumor-bearing mice but be restricted by tumor size. The results suggest that transfecting the B7 gene into cervical cancer as a cell vaccine may be an efficient supplementary method to treat cervi cal cancer after operation.     

CD40L融合改造后的人乳头瘤病毒16型E7基因DNA疫苗的构建及其免疫原性测定

目的研究针对人乳头瘤病毒(HPV)16型的DNA疫苗,测定其免疫原性,用于治疗HPV16感染及感染相关恶性肿瘤。方法联合采用基因切割重排、定点突变及密码子优化等策略改造HPV16的转化基因E7,基因合成法获得改造后的E7基因(mE7);PCR法将mE7基因与CD40L胞外区编码序列融合,然后以pVR1012为载体构建pVR1012-mE7(mE7)及pVR1012-mE7/CD40L(mE7/CD40L)表达质粒;经肌肉免疫C57BL/6小鼠;ELISA法检测E7特异性血清抗体水平,ELISPOT法分析E749-57(H-2b)特异性分泌IFN-γ的CD8 T细胞活化水平,胞内染色-流式细胞检测分析E7特异性CD4 Th细胞活化水平;并在C57BL/6小鼠体内进行疫苗抗瘤活性检测。结果与野生型E7基因(wE7)相比,mE7基因诱发产生的E7特异抗体水平(P<0.01)、分泌IFN-γ的CD8 T细胞数目(P<0.01)及CD4 Th细胞活化水平(P<0.05)均显著提高;与mE7基因相比,mE7/CD40L融合基因可进一步显著提高E7特异性分泌IFN-γ的CD8 T细胞数目(P<0.01),但对E7特异性抗体产生及CD4 Th细胞活化水平没有明显影响。疫苗小鼠体内预防性免疫实验中,经wE7免疫的小鼠接种瘤细胞后2周内全部形成移植瘤,而所有经mE7及mE7/CD40L免疫的小鼠在瘤细胞攻击后第7周仍未见移植瘤形成;疫苗体内治疗性免疫实验中,小鼠在接种wE7后第8天左右全部形成移植瘤,并呈渐进性生长,而接种mE7的小鼠移植瘤清除率为30%,接种mE7/CD40L的小鼠移植瘤清除率增高至45%。对移植瘤的组织学检查结果显示,mE7/CD40L及mE7免疫组小鼠瘤细胞间及瘤组织周围可见大量淋巴细胞浸润,而wE7组小鼠的瘤细胞呈编织状紧密排列,未见有淋巴细胞浸润。结论HPV16型mE7/CD40L融合基因疫苗免疫小鼠后可诱发较强的E7特异性细胞免疫及体内抗瘤活性,具有较强的免疫原性。     

厌氧棒状杆菌菌苗对S_(180)抗癌效应的研究

本文在CFW小鼠S-180实验研究中,证明给小鼠右腋部皮下接种S-180瘤细胞后,随着肿瘤生长,动物腹腔细胞和局部淋巴结细胞的CMC和ADCC活性都下降,厌氧棒状杆菌菌苗治疗能抑制小鼠肿瘤生长,同时增强腹腔细胞和淋巴细胞的CMC和ADCC活性。     

Th17细胞在小鼠H22原位肝癌模型中的表达和意义

目的 检测小鼠原位肝癌模型中Th17细胞的表达,探讨其在肝癌免疫中的作用. 方法 30只Balb/C小鼠随机分为肝癌模型组（10只）、生理盐水对照组（10只）、空白对照组（10只）,流式细胞术检测小鼠分离纯化后的各组新鲜脾脏Th17细胞的相对比例;实时RT-PCR检测各组肝组织IL-17基因和RORγT基因表达水平.结果 肝癌模型组脾脏Th17/ CD4＋T细胞的比例高于生理盐水对照组脾脏、空白对照组脾脏,差异有统计学意义（P 〈 0.01）,而各对照组之间的比例差异无统计学意义（P 〉 0.05）.肝癌模型组肿瘤组织IL-17 mRNA和RORγT mRNA的表达水平均高于各自对照组,差异有统计学意义（P 〈 0.01）,而IL-17 mRNA和RORγT mRNA在各自对照组之间表达水平的差异无统计学意义（P 〉 0.05）. 结论 Th17细胞在小鼠原位肝癌模型中数量增多,IL-17表达升高,增强免疫反应,但在肝癌免疫中的作用及机制复杂,有待深入研究.     

Induction of protective antitumor activity of tumor lysate-pulsed dendritic cells vaccine in RM-1 prostate cancer model

Objective： To investigate the antitumor activity of tumor lysate-pulsed dendritic cells vaccine in RM-1 prostate cancer mice model with the survival time of mice calculated and the tumor size measured in DC vaccine therapy. Methods： C57BL/6 mice were immunized on the dorsal flank by s.c. inoculation of Lysate-DC, ova-DC, and non-DC on day -7. On day 0, 2× 10^6cells of RM-1 tumor cells （H-2b） were injected s.c. in C57BL/6 mice pre-treated by s.c. inoculation of modified DCs, correspondingly. DTH assay was performed with modified DCs. In partial test, for the determination of which immune cells were required for antitumor activity, mice were immunodepleted of CD4, CDS, or natural killer （NK） NK1.1 cells with the corresponding monoclonal antibodies. The survival time of nude mice loaded with tumor cells was calculated and the size of tumor measured. Results： In RM-1 mice prostate cancer model, immunized with lysate-DC, compared with ova-DC and non-DC, the pre-infection vaccine resulted in 100% clearance of primary tumors, whereas on day 0 of injection vaccine cleared 40-60% of primary tumors. On day 0, C57BL/6 mice （H-2b） were immunized with Lysate-DC, compared with ova-DC and non-DC by caudal vein injection, then on day 15, RM-1 cells were inoculated. On day 30, average diameters of tumor in different groups of modified DC were 23.7±5.4 mm, 22.1±4.9 mm, 4.3±2.6 mm, respectively. Lysate-DC, compared with ova-DC and non-DC, can greatly depressed RM-1 tumor cell growth （P〈0.01）. The mean survival time of C57BL/6 mice in Lysate-DC, ova-DC and non-DC groups were 15.8±2.6, 16.6±3.2, 39.0±5.6, respectively, and there was a significant difference in the mean survival time in lysate-DC group between ova-DC and non-DC group （P〈0.01）. DTH test showed that lysate-DC could prime T lymphocyte and elicit tumor antigen specific immune response, and over 80% mice in groups of lysate-DC showed obvious swelling in their foot pad. This response was strengthened with repeating inoculation, whereas DTH response was not seen in control group. In vivo depletion of NK cells resulted in a 40-60% reduction in growth suppression within the primary tumor, and depletion of CD4^＋ cells resulted in a 20% reduction in growth suppression. Conclusion： The minor lysate-pulsed dendritic cells vaccine could elicit antitumor activity in RM-1 loaded C57BL/6 mice, and prolong the duration of RM-1 loaded C57BL/6 mice. So DC-based immunotherapy with hormone-refractory prostate carcinoma yielded protective immunity, generated efficient cellular antitumor responses, thereby providing further preclinical support for feasible immunotherapy approaches for prostate cancer.     

Splenectomy suppresses growth and metastasis of hepatocellular carcinoma through decreasing myeloid-derived suppressor cells <Emphasis Type="Italic">in vivo</Emphasis>

The function of the spleen in tumor development has been investigated for years. The relationship of the spleen with hepatocellular carcinoma (HCC), a huge health burden worldwide, however, remains unknown. The present study aimed to examine the effect of splenectomy on the development of HCC and the possible mechanism. Mouse hepatic carcinoma lines H22 and Hepa1-6 as well as BALB/c and C57 mice were used to establish orthotopic and metastatic mouse models of liver cancer. Mice were divided into four groups, including control group, splenectomy control group (S group), tumor group (T group) and tumor plus splenectomy group (T+S group). Tumor growth, metastases and overall survival were assessed at determined time points. Meanwhile, myeloid-derived suppressor cells (MDSCs) were isolated from the peripheral blood (PB), the spleen and liver tumors, and then measured by flow cytometery. It was found that liver cancer led to splenomegaly, and increased the percentage of MDSCs in the PB and spleen in the mouse models. Splenectomy inhibited the growth and progression of liver cancer and prolonged the overall survival time of orthotopic and metastatic models, which was accompanied by decreased proportion of MDSCs in the PB and tumors of liver cancer-bearing mouse. It was suggested that splenectomy could be considered an adjuvant therapy to treat liver cancer.     

小鼠脾来源树突状细胞与NS_1骨髓瘤细胞融合体的抗肿瘤作用

目的：观察ＢＡＬＢ／Ｃ小鼠脾来源树突状细胞（ｓｐｌｅｅｎｄｅｎｄｒｉｔｉｃｃｅｌｓ,ｓＤＣ）与肿瘤细胞融合体的抗肿瘤效应。方法：以灭活的ＮＳ１细胞免疫活化ＢＡＬＢ／Ｃ小鼠,取其ｓＤＣ与ＮＳ１骨髓瘤细胞融合,并筛选出融合细胞;用此融合细胞作为瘤苗,免疫治疗皮下荷ＮＳ１瘤的小鼠２次,间隔１周。结果：融合细胞瘤苗本身无致瘤性。用融合细胞瘤苗免疫治疗荷瘤小鼠后,其瘤体消失,且在观察期６０ｄ内未见肿瘤转移和复发。结论：脾来源树突状细胞与ＮＳ１细胞融合瘤苗免疫荷瘤小鼠后,激发其体内存在的抗ＮＳ１肿瘤效应。     

重组免疫毒素EGF-TCS对荷瘤小鼠抗肿瘤效果的实验研究

目的 使用EGF-TCS重组蛋白对裸鼠的肿瘤模型进行体内抑瘤实验,观察其抗肿瘤效果.方法 分别给裸鼠右腋皮下接种人肝癌BEL-7402细胞,接种后第6天,尾静脉注射100、50和25 μg/kg EGF-TCS进行治疗,设空白对照组,停药后第2天处死小鼠,称量瘤重,计算抑瘤率;肿瘤组织做免疫组织化学实验,从而研究其抑制肿瘤生长的途径.结果 高、中和低剂量组的抑制率分别为71.3%、60.87%和45.22%,对治疗结束后各组平均瘤重进行统计学分析,结果有显著性差异(F=8.712,P=0.006);免疫组化结果显示EGF-TCS可以明显的抑制肿瘤血管的形成,高剂量组肿瘤组织未有可见血管,中低剂量组肿瘤组织血管明显少于模型组,其抑制肿瘤生长的途径可能是通过抑制肿瘤血管生长从而达到治疗目的.结论 EGF-TCS可以有效地抑制荷瘤小鼠实体瘤的生长,预示了该免疫毒素在肿瘤治疗中的潜在应用价值.