OK-432对荷瘤小鼠脾细胞IL-12及Th1细胞因子分泌的诱导作用

目的：探讨溶血链球菌冻干制剂OK－432的抗肿瘤机理。方法：B16黑色素瘤细胞接种于C57BL／6小鼠皮下,3d后腹腔注射1KU的OK－432,每周1次,连续3周。观察肿瘤生长体积及荷瘤小鼠的生存期,并测定了OK－432在体内、体外对荷瘤脾细胞IL－2、IL－4,IL－6,IL－10,IL－12,IFN-γ分泌的结果。结果：OK－432能显著抑制B16黑色素瘤的生长,延长荷瘤小鼠的生存期（P＜0．05）;在体外OK－432可刺激荷瘤小鼠脾细胞IL－6,IL－10,IL－12,IFN-γ的分泌（P＜0．01）;腹腔注射OK－432后荷瘤小鼠脾细胞IL－2,IL－12,IFN-γ的分泌显著增加,而IL－10显著减少（P＜0．05）。提示OK－432治疗后荷瘤小鼠脾细胞Th1增加,Th2相对减少。结论：OK－432在体内可通过诱导荷瘤小鼠脾细胞IL－12的分泌,促进Th0向Th1分化,使宿主的免疫功能处于Th1优势状态,从而增强宿主的抗肿瘤作用。     

不同辐射剂量损伤后T淋巴细胞亚群及Th1和Th2的变化

目的： 观察不同辐射剂量损伤对小鼠T淋巴细胞功能亚群的影响,探讨辐射所致的免疫系统损伤在细胞学和分子水平上的机制。方法：C57BL/6j小鼠采用60 Coγ射线照射诱导辐射损伤模型。总照射剂量分别为0、0.7、1.4、2.8和5.6 Gy。应用细胞表面标志和细胞内因子标记,流式细胞仪检测分析不同剂量组小鼠脾T淋巴细胞功能亚群CD3＋、CD4＋、CD8＋ T细胞改变及Th1和Th2的变化。结果：与空白对照组比较,小鼠脾T淋巴细胞功能亚群CD3＋、CD4＋、CD8＋ T细胞在照射后均明显降低(P＜0.01),降低幅度与受照剂量明显相关。CD4＋/CD8＋比值显示照射后均明显升高(P＜0.01)。照射后Th1水平均降低(P＜0.01),而在照射剂量≥1.4 Gy时Th2均明显降低(P＜0.01),其中Th1降低明显高于Th2。受照剂量大于2.8 Gy组Th2/Th1比值较空白对照组明显升高(P＜0.01)。结论：不同照射剂量对T淋巴细胞功能亚群CD3+、CD4+、CD8+ T细胞和CD4＋/CD8＋比值有明显影响,Th1,Th2和Th2/Th1功能亚群的失衡在辐射所致的免疫损伤中具有重要作用。     

微波固化保肢术对骨肉瘤患者Th1/Th2漂移的逆转

目的：研究微波组织固化保肢技术对患者免疫功能的影响。方法：应用放射免疫分析法以及酶联免疫分析法检测20例行微波组织固化保肢术组骨肉瘤患者及20例其它手术组患者手术前后血清IL－2,IL－4,IL－6,IL－10,IL－12的含量,应用流式细胞仪检测患者淋巴细胞亚群的变化。结果：与正常对照相比,骨肉瘤患者CD4^ T细胞,血清中IL－2,IL－12减少（P＜0．01）,CD8^ T细胞,IL－10,IL－4,IL－6含量升高（P＜0．01）,CD4／CD8比值降低（P＜0．01）,患者免疫功能受到抑制;术后30天截肢组及保肢组CD4^ T细胞,血清中IL－2,IL－12均升高,CD8^ T细胞,IL－10,IL－4,IL－6含量降低（P＜0．01）,CD4／CD8比值升高（P＜0．01）,患者免疫功能较术前改善;与截肢组比较,保肢组患者CD4^ T细胞、IL－2,及CD4／CD8比值30天上升幅度明显高于截肢组（P＜0．01）,CD8^ T细胞,IL－4,IL－6 30天降低幅度明显高于截肢组（P＜0．01）。IL－2,IL－12与CD4^ 细胞成正相关,IL－4,IL－6,IL－10与CD4^ T细胞成负相关。结论：微波固化保肢术可以加速逆转骨肉瘤患者Th1/Th2和Th2漂移,增强患者的免疫功能。     

免疫重建荷人高转移肝癌SCID鼠模型的建立及其鉴定

目的：建立具有人免疫学特性的高转移肝癌SCID鼠模型。方法：SCID小鼠腹腔注射人外周血淋巴细胞,皮下接种人高转移肝癌细胞MHCC97-H,免疫重建人高转移肝癌SCID小鼠模型,并鉴定建立的结果。结果：①免疫重建荷人高转移肝癌细胞小鼠的成瘤率为100％,较荷瘤组成瘤潜伏期延长,体积缩小（P〈0．05）;转移率为100％,其从皮下转移到肝脏所需时间较荷瘤组延长（P〈0．05）。②第2、4、6周小鼠血中人IgG含量的测定：同期相比,人化荷瘤组小鼠血中人IgG含量高于人化组（P〈0．05）。③第6周SCID小鼠外周血中人CD3^＋T淋巴细胞和人CD20^＋B淋巴细胞含量的测定：人化荷瘤组小鼠血中人CD3^＋T淋巴细胞和人CD20^＋B淋巴细胞含量高于人化组（P〈0．05）。④免疫组化检测人化荷瘤组小鼠脾脏中存在人CD3^＋T淋巴细胞和CD20^＋B淋巴细胞。结论：成功建立免疫重建荷人高转移肝癌SCID鼠模型,为肝癌转移的研究及治疗提供了理想的动物模型。     

肺癌患者红细胞免疫功能与外周血T淋巴细胞亚群的改变

目的：观察肺癌患红细胞免疫功能及T淋巴细胞亚群的变化。方法：应用红细胞免疫粘附花环形成法和流式细胞仪检测法。结果：（1）肺癌组（55例）红细胞C3b受体花环率（RBC－C3bRR)、CD3^ 、CD4^ 、CD^ ／CD^8 比值低下正常人（20例）（P＜0．05－0．01）,红细胞免疫复合物花环率（RBC－ICR）依次高于肺良性肿瘤组（15例）和正常人（P＜0．05－0．01）。（2）肺癌RBC－C3bRR与CD4^ /CD8^ 成直线正相关（P＜0．01,r=0.9131)。（3）Ⅲa,Ⅲb期肺癌,RBC－C3bRR,CD3^ ,CD4^ ,CD4^ /CD8^ 比值低于Ⅰ、Ⅱ期肺癌患和正常人（P＜0．05－0．01）,而RBC－ICR高于正常人（P＜0．01）。Ⅰ、Ⅱ期病人CD3^ ,CD4^ ,CD8^ 低于正常人（P＜0．01）,RBC－ICR高于正常人（P＜0．01）。结论：肺癌患RBC免疫和T细胞亚群的测定及其相关性分析,对于肺癌的诊断、治疗及病情预后估计有一定价值。     

化疗对晚期肺癌患者T淋巴细胞亚群和红细胞免疫功能的影响

目的：分析42例晚期肺癌者化疗前后T淋巴细胞亚群和红细胞免疫功能的变化,并探讨其与病情的关系。方法：对42例晚期肺癌化疗前后的血标本采用流式细胞术检测T淋巴细胞亚群和采用交体粘附法检测红细胞免疫功能,并与体检健康者作比较,结果：本组晚期肺癌患者治疗前后T淋巴细胞亚群和红细胞免疫功能均低于对照组（P＜0．05,P＜0．01）。治疗后,化疗有效率总T细胞（CD3^ ）、辅助／诱导T淋巴（CD4^ ）、CD4^ 与CD8^ 比值均显著升高（P＜0．01）,细胞毒／抑制性T淋巴细胞（CD8^ ）降低（P＜0．05）;化疗无效者CD3^ 、CD4^ 、CD4^ ／CD8^ 显著降低（P＜0．05,P＜0．01）,而CD8^ 升高（P＜0．01）,直向肿瘤红细胞花环（direct tumor erythrocyte rosette,DTER）,红细胞C3b受体花环（red blood cell C3b receptor rosette,RBC-C3bRR)和红细胞免疫复合物花环（red blood cell immunity complex rosette,RBC-ICR),化疗前后变化不明显（P＞0．05）。结论：晚期肺癌患者免疫功能低下,有铲化疗能提高患者的T淋巴细胞亚群免疫功能,临床上对免疫功能的观察对肺癌患者的治疗和预后有一定的监测作用。     

老年肺癌患者血清T淋巴细胞亚群CD28的检测及临床意义

目的：探讨老年肺癌患者淋巴细胞免疫功能状态、临床意义及与肺癌病理类型、临床分期的关系。方法：采用三色免疫荧光标记流式细胞术检测35名正常老年人及32例老年肺癌患者外周血的总T淋巴细胞（CD3^＋）、辅助／诱导T淋巴细胞（CD4^＋）、抑制／细胞毒T淋巴细胞（CD8^＋）、细胞毒T细胞（CD8^＋CD28^＋）和抑制T细胞（CD8^＋CD28）。结果：老年肺癌组CD4^＋Tz细胞（t=2．01,P〈0．05）和CD3^＋、CD8^＋T细胞及CD8^＋CD28^＋T细胞亚群明显低于老年正常对照组,t=2．01,P〈0．01;CD8^＋CD28T细胞亚群明显高于老年正常对照组,t=2．01,P〈0．01;老年肺癌患者中,T淋巴细胞及亚群CD28的表达在TNM临床分期及病理类型间差异均无统计学意义,t=2．11,P值均〉0．05。结论：老年肺癌患者存在明显的T淋巴细胞亚群免疫功能紊乱,抗肿瘤能力明显下降,并导致病情恶化更严重,且与病理类型及临床分期无关。     

HepA-H和HepA-L肝癌荷瘤小鼠CD8+/CD28+T细胞和肿瘤新生淋巴管的研究

目的研究不同淋巴转移潜能肝癌（HepA-H和HepA-L）荷瘤小鼠体内CD8^＋／CD28^＋T淋巴细胞和肿瘤新生淋巴管，探讨与肿瘤淋巴转移的关系。方法用两种来源相同，而淋巴转移能力不同的小鼠肝癌细胞亚系，HepA-H（高淋巴转移）和HepA-L（低淋巴转移），小鼠皮下接种，制备荷瘤动物模型。采用抗小鼠CD8、CD28荧光标记单克隆抗体，流式细胞术定量检测小鼠外周血和肿瘤组织内CD8^＋／CD28^＋T淋巴细胞数量；应用5＇-核苷酸酶-碱性磷酸酶双重组化染色法，观察肿瘤新生淋巴管。结果肿瘤皮下接种后14天，HepA-H组荷瘤小鼠外周血CD8^＋／CD28^＋T淋巴细胞数量明显低于正常对照组（P〈0．05）HepA-H组肿瘤组织内CD8^＋／CD28^＋T淋巴细胞数量显著低于HepA-L组（P〈0．05）。两种肿瘤细胞亚系均可诱导肿瘤新生淋巴管，但HepA-H肿瘤内和瘤周最高淋巴管密度均明显高于HepA-L组（P〈0．01）。结论肿瘤淋巴转移能力的差异与体内CD8^＋／CD28^＋T淋巴细胞数量及肿瘤新新生淋巴管有关。     

CD4+T细胞在抗肿瘤过继性免疫治疗中作用的研究

目的：观察CIK细胞中CD4^＋T细胞中Th1／Th2亚群分布情况。方法：体外大规模扩增CIK细胞，磁珠法纯化其中CD4^＋T细胞．采用胞内染色、ELISA和荧光定量RT—PCR法检测培养前后Th1／Th2细胞亚群的比例、Th1／Th2类细胞因子的分泌量和基因表达的变化。结果：富集CD4^＋细胞纯度达96％，CIK中Th1亚群和Th0亚群的比例较PBMC显著升高（P〈0．05），而Th2亚群无显著变化。CD4^＋CIK细胞中IL-2和IFN-γ等Th1类细胞因子释放量增高（P〈0．01），而IL－10和TGF-β等Th2类细胞因子释放量降低（P〈0．05）。但CD4^＋CIK细胞中除IFN-γ／的mRNA显著升高外（P〈0．05）．IL-2和IL-4的mRNA变化不明显。结论：CIK细胞中的CD4^＋T细胞具有明显的Th1优势。     

肝癌荷瘤小鼠调节性T细胞数量的改变及其与肿瘤生长的关系

目的研究肝癌荷瘤小鼠调节性T细胞数量的改变及其与肿瘤生长的关系。方法采用小鼠肝癌细胞系H22接种BALB／c小鼠,建立肝癌模型;采用流式细胞术方法检测CD4^＋ CD25^＋T／CD4^＋T细胞的比例;以RT-PCR和流式细胞术检测Foxp3基因的表达。以免疫磁珠分选法纯化CD4^＋CD25^＋T和CD4^＋CD25^-T细胞;在体外,用3H-TdR掺入法检测T细胞的增殖情况;在体内,观察荷瘤小鼠来源的CD4^＋CD25^＋T细胞对肿瘤生长的作用。结果（1）荷瘤小鼠在引流淋巴结中,CD4^＋CD25^＋T细胞占CD4＋T细胞（18．80％±0．06％）比例增高,与对照组（9．50％±0．03％）相比,差异有统计学意义（P〈0．01）;在非引流淋巴结（LN）和脾脏（SP）中,荷瘤小鼠CD4^＋CD25^＋T／CD4^＋T比例分别为16．28％±0．02％和17．28％±0．06％,与对照组9．50％±0．03％和11．08％±0．04％相比,差异有统计学意义（P〈0．01,P〈0．05）;同时,调节性T细胞特异性标志Foxp3 mRNA的表达也升高。在同一只荷瘤小鼠中,引流淋巴结中CD4^＋CD25^＋T细胞数量（18．8％±0．06％）较对侧非引流淋巴结（16．28％±0．02％）略有升高,但差异无统计学意义（P〉0．05）。（2）从荷瘤小鼠中纯化的CD4^＋CD25^＋T细胞,在体外对抗CD3单抗的刺激无反应,但能抑制CD4^＋CD25^-T细胞的增殖。（3）CD4^＋CD25^＋T／CD4＋T比例与肿瘤大小呈正相关,并且可以抑制CD4^＋CD25^-T细胞的抗肿瘤效应。结论肝癌细胞在小鼠体内的生长可以提高调节性T细胞的数量,其数量的高低与肿瘤的大小呈正相关,提示清除调节性T细胞将是肿瘤免疫治疗的策略之一。     

IL-6对受照荷瘤小鼠正常组织的辐射防护效应

目的　以荷瘤小鼠为观察对象 ,研究白细胞介素 6 (Interleukin - 6 ,IL - 6 )对受照射荷瘤小鼠正常组织的辐射防护作用。方法　本实验用黄嘌呤氧化酶法测定受全身照射的荷瘤小鼠肝、肾、脑、肺组织中的总超氧化物歧化酶 (T -SOD)含量 ;用硫代巴比妥酸钠 (TBA)荧光法测定以上组织中的脂质过氧化物 (LPO)产物丙二醛 (MDA)的含量 ;双底物偶联动力学法测定以上组织中的谷光苷肽过氧化物酶 (GSH -PX)含量。结果　对于接受全身照射的荷瘤小鼠IL - 6可提高肝、脑组织中的抗氧化酶T -SOD、GSH -PX活性 (P <0 .0 1,P <0 .0 5 ) ,降低肝、脑组织中MDA含量 (P <0 .0 5 )。结论　IL - 6对受全身照射的荷瘤小鼠可肝、脑组织有辐射防护作用。     

Selective suppression of the generation of anti-tumor L3T4+ but not of Lyt-2+ T cell-mediated immunity in the tumor-bearing state

C3H/He mice hyperimmune against syngeneic MH134 hepatoma were prepared by intradermal (id) inoculation of viable tumor cells followed by surgical resection of the tumor and by repeated id challenges with viable tumor cells. Winn assays performed utilizing spleen cells from these mice have revealed that both Lyt-2+ and L3T4+ T cell subsets from MH134-hyperimmune mice produced complete tumor protection. The in vivo tumor-neutralizing activity was also found in spleen cells from tumor-bearing mice at various times after id implantation of MH134 tumor cells. However, in contrast to comparable tumor-neutralization by Lyt-2+ and L3T4+ T subsets from hyperimmune mice, only the Lyt-2+ T cell subset from tumor-bearing mice was capable of mediating the in vivo protective immunity. L3T4+ T cell-mediated immunity was not detectable in the tumor-bearing state irrespective of the length of the sensitization period with a primary growing tumor, but emerged in the mice which resisted the first tumor challenge after the resection of the primary tumor. These results indicate that the emergence of L3T4+ T cell-mediated anti-tumor immunity is stage-dependent and the Lyt-2+ T cells represent the main functional subset in the tumor-bearing state, although both subsets of T cells are potentially capable of effecting anti-tumor in vivo immunity. The results are discussed in relation to the selective suppression of the L3T4+ but not of Lyt-2+ T cell function in the tumor-bearing state.     

蚕丝蛋白肽对S180荷瘤小鼠免疫调节作用的研究

目的:探索蚕丝蛋白肽(silk fibroin peptide,SFP)在肿瘤环境下的免疫调节作用,为临床应用提供药理学基础.方法:构建小鼠S180皮下移植瘤模型,分为空白对照组(生理盐水),阳性对照药胸腺五肽组(TP-5,1 mg/kg),蚕丝蛋白肽低剂量组(SFP-L,300 mg/kg)和蚕丝蛋白肽高剂量组(SF...     

Effects of cancer immunotherapy with indomethacin and interleukin-2 on murine hemopoietic stem cells.

We examined: (a) whether in vitro-generated lymphocyte-activated killer (LAK) cells from normal mice and splenic killer cells from tumor-bearing mice subjected to interleukin-2 (IL-2) therapy alone or in combination with chronic indomethacin therapy have any detrimental effects on the spleen colony-forming units (CFU-S) of the normal bone marrow (BM); and (b) the effects of these immunotherapy protocols on CFU-S numbers in host hemopoietic organs. Effects of in vitro-generated LAK cells (normal C3H/HeN mouse splenocytes cultured with 1000 units IL-2/10(6) cells for 72 h) on BM CFU-S were examined by incubating macrophage-depleted BM cells with LAK cells at 1:2.5 and 1:5 BM:LAK cell ratios or with LAK cell supernatant for 4 h. The cells were washed and subsequently injected into irradiated mice. Irradiated mice were also reconstituted with BM cells or LAK cells incubated alone. Spleen colonies were scored macroscopically and microscopically on day 7 after reconstitution of lethally irradiated mice with the various cell combinations. A comparison of colony numbers produced by LAK and BM cell mixture revealed that LAK cells at either dose had no suppressive effect on the colony-forming ability of BM at the macroscopic and microscopic levels of analysis. The supernatant of cultured LAK cells had a minor suppressive effect on colony formation at the macroscopic but not the microscopic level of analysis, indicating the presence of one or more suppressive factors capable of mediating a short-term inhibitory effect. In the immunotherapy experiment, C3H/HeN mice transplanted s.c. with 5 x 10(5) C3L5 mammary adenocarcinoma cells received either vehicle alone (controls), IL-2 (1.5 x 10(4) Cetus units i.p. every 8 h on days 10-14 and days 20-25), or chronic indomethacin therapy (10 micrograms/ml in drinking water from day 5 onwards) plus IL-2 as above. Animals were killed 24-25 days after tumor transplantation to examine: (a) the number of metastatic lung nodules; (b) the effects of co-incubating therapy-generated splenic effector cells with normal BM cells for 4 h on BM CFU-S, and (c) the CFU-S content of host BM and spleen. Results revealed a drop in spontaneous lung metastases from a mean of 50 in control mice to 18 with IL-2 therapy alone, and to 5 with chronic indomethacin therapy plus IL-2 therapy. Splenocytes from normal and tumor-bearing control or treated mice, when incubated with normal BM, had no effect on spleen colony formation at the macroscopic level.(ABSTRACT TRUNCATED AT 400 WORDS)     

补康灵对辐射损伤肺癌小鼠免疫功能影响的观察

目的:观察补康灵对辐射损伤肺癌小鼠免疫功能的防护作用。方法:C57BL/6小鼠随机分组,制备Lewis肺癌模型后,一次性接受4.0Gy/只60 Coγ射线照射,补康灵组连续给药10d,其他组给予等量的生理盐水,检测小鼠的胸腺指数、脾指数、T淋巴细胞亚群、IL-2和IL-6等。结果:与辐射组比较,补康灵中、高剂量联合辐射组小鼠的提质量显著增长,P<0.05;补康灵中、高剂量组肺癌小鼠血清IL-2分别为(21.78±1.92)和(24.05±1.65)ng/L,IL-6分别为(39.22±5.23)和(42.08±6.52)ng/L,CD4+/CD8+分别为1.37±0.31和1.59±0.33,均明显提高;胸腺和脾脏指数分别为22.28±6.95、26.23±7.01和47.82±8.32、51.20±9.19,差异有统计学意义,P<0.05。结论:补康灵对辐射所致的小鼠免疫功能损伤具有防护作用。     

Changes in peripheral lymphocyte subsets in patients after partial microwave ablation of the spleen for secondary splenomegaly and hypersplenism: a preliminary study.

PURPOSE: Microwave ablation therapy for secondary splenomegaly and hypersplenism has been shown to be effective from pre-clinical animal models and clinical investigations. This study was performed to determine its effects on the status of peripheral lymphocyte subsets in patients receiving microwave ablation of the spleen. MATERIALS AND METHODS: Ten patients with secondary splenomegaly and hypersplenism received microwave ablation of the spleen during laparoscopy or percutaneously under ultrasound guidance. The percentage peripheral blood T cells, B lymphocytes and NK cells were measured using flow cytometry before and on days 1, 3 and 7 after therapy, as well as 1 and 3 months afterwards. RESULTS: Percentages of CD3(+) and CD4(+) cells increased rapidly 1 month after therapy. There was no significant change in CD8(+), CD4(+)/CD8(+) or NK cells of the pre- and post-therapy levels and B lymphocytes increased significantly after therapy. In patients with an ablation volume (AV) less than 20% (group A), T cells increased 1 month after ablation but decreased 3 months after ablation. B lymphocytes increased significantly after surgery. Levels of NK cells were lower than that before therapy on each testing. In patients with 20-40% AV (group B), levels of T cells, B lymphocytes and NK cells showed an increase. Levels of CD4(+) cells were significantly higher in group B than in group A, 3 months after therapy. CONCLUSIONS: Microwave ablation therapy for splenomegaly and hypersplenism appears to have a favourable effect on peripheral lymphocyte subsets. A relationship may exist between the ablation volume and the level of peripheral lymphocyte subsets.     

蛋氨酸脑啡肽联合低剂量纳曲酮治疗胰腺癌的实验研究

目的:通过单独应用MENK、NTX 与联合应用MENK 与NTX 的体内外实验,探讨MENK、NTX 及二者联合时对胰腺癌的抑制作用。方法:体外实验中,MTS方法检测各实验组中胰腺癌增殖细胞的抑制情况;体内实验中,通过建立胰腺癌荷瘤小鼠模型,检测各实验处理组中胰腺癌的抑制情况。 WB法检测荷瘤小鼠胰腺癌组织阿片受体蛋白表达量;流式细胞术检测荷瘤小鼠淋巴细胞亚群（CTL、Treg、NK、γδT）的增殖情况。结果:在体外实验中,MENK 在浓度为10-6 mol/L时对胰腺癌细胞的抑制作用最强;NTX在浓度为10-5 mol/L时对胰腺癌细胞的抑制作用最强;二者相比较,MENK的作用更加显著,具有统计学意义（P＜0.05）;在体内实验中,MENK 与NTX 联合应用在48 小时后与对照组相比较,具有统计学意义（P＜0.05）,在96小时,与MENK组和NTX 组相比较抑癌率升高,具有统计学意义（P＜0.05）;在体内实验中,在1次/24 h的给药方式下,MENK在剂量为20 mg/kg 时抑癌效果最显著;NTX 在剂量为5 mg/kg 时抑癌效果最显著;MENK组与NTX组相比较作用更加显著,具有统计学意义（P＜0.05）;MENK 与NTX 联合应用时,MENK 在剂量为20 mg/kg 1 次/24 h 与NTX 在剂量为5 mg/kg 1 次/96 h 给药方式时,抑癌效果最显著,与对照组相比具有统计学意义（P＜0.05）;Western-blot 技术检测荷瘤小鼠肿瘤组织阿片受体的蛋白的表达含量,NTX 组与MENK+NTX 组肿瘤组织阿片受体蛋白表达量增加,NTX 组高于MENK+NTX 组,具有统计学意义（P＜0.05）;流式细胞技术检测荷瘤小鼠淋巴细胞亚群增殖情况,MENK组、NTX 组与MENK+NTX组都可以促进淋巴细胞亚群（CTL、NK、γδT）增殖,抑制Treg增殖,与对照组相比具有统计学意义（P＜0.05）。结论:MENK在体内外能够抑制胰腺癌细胞增殖,NTX在体内外能够抑制胰腺癌细胞增殖,两者相比MENK作用更加显著;MENK与NTX联合应用,在NTX给药72小时后具有协同作用;NTX在体内能够上调肿瘤组织阿片受体的蛋白表达;MENK和NTX均能在体内能够促进CTL、NK、γδT细胞的增殖,抑制Treg细胞的增殖,二者联合具有协同作用。     

Restoration of impaired T cell functions in tumor-bearing mice by the administration of interleukin 1

The effect of in vivo administration of recombinant human interleukin 1 (IL-1) on T cell functions in tumor-bearing mice was studied using an in vitro assay system. The in vitro induction of trinitrophenyl (TNP)-specific cytotoxic T cell and proliferative T cells responses from spleen cells was impaired in X5563 plasmacytoma-bearing C3H/He mice. However, the administration of IL-1 alpha or IL-1 beta to tumor-bearing mice restored T cell functions in a dose-dependent manner. Antigen-presenting activities of spleen cells in tumor-bearing mice for T cell activation were not restored by the administration of IL-1. The activities of cytotoxic T cells and cytostatic T cells specific for X5563 cells were also enhanced by the administration of IL-1. Furthermore, in IL-1-treated mice, NK cell activity of spleen cells detected in terms of the killing of Yac-1 cells was also restored. In accordance with these results, the growth of X5563 cells was significantly inhibited and the lymphocytes from IL-1-treated mice specifically inhibited the growth of tumor cells. These results suggest that the in vivo administration of IL-1 restored the impaired T cell and NK cell functions in tumor-bearing mice and activated protective immunity against tumor cells. Thus, recombinant IL-1 can be applied for tumor immunotherapy.     

转染CD40配体的卵巢癌细胞株OVHM抗卵巢癌肝转移机制的研究

目的 探讨转染CD40配体(CIMOL)的卵巢癌细胞株OVHM(CD40L-OVHM)体内抗卵巢癌肝转移的可能机制.方法 6～8周龄的C57BL/6N×C3H/He杂交一代(B6C3F1)雌性小鼠5只,经小鼠脾脏内接种OVHM,应用HE染色法验证小鼠肝脾转移模型是否建立成功.将OVHM细胞、空载体DNA-pMKITneo-OVHM细胞和CD40L-OVHM细胞接种于B6C3F1小鼠的脾脏,应用流式细胞术,分析荷瘤小鼠脾细胞CD11c分子的表达情况;应用四甲基偶氮唑蓝(MTT)法,检测荷瘤小鼠脾脏细胞毒性T淋巴细胞(CTL)的杀伤活性;应用酶联免疫吸附试验(ELISA),检测荷瘤小鼠外周血血清中干扰素(IFN)-γ、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-12、IL-4和IL-10的含量,并且观察小鼠脾脏和肝脏的成瘤情况及小鼠生存时间.结果 经病理学证实,卵巢癌肝脾转移动物模型建立成功.CD40L-OVHM组中小鼠脾细胞CD11c阳性细胞率明显高于OVHM组和转染空载体DNA-pMKITneo-OVHM组.CD40L-OVHM组小鼠脾脏内CTL的特异性杀伤活性明显增强,小鼠外周血血清中IFN-γ、TNF-α和IL-12的含量明显升高,而IL-4和IL-10的含量则明显降低,与OVHM组和转染空载体DNA-pMKITneo-OVHM组相比,差异均有统计学意义(P<0.05).CD40L-OVHM组小鼠的肝脏和脾脏重量均明显低于OVHM组和转染空载体DNA-pMKITneo-OVHM组,并且小鼠的生存期也明显延长(P<0.05).结论 转染CD40L cDNA的卵巢癌细胞可促进脾脏树突状细胞(DC)的成熟和分化,增强脾脏CTL的特异性杀伤活性,诱导Th1型细胞因子的分泌,抑制Th2型细胞因子的分泌,这可能是CD40L基因体内抗卵巢癌肝转移的机制之一.     

The changing pattern of the splenic lymphocyte subsets in tumor-bearing mice after oral treatment with OK-432.

The aim of this study was to investigate the influence of oral administration of OK-432 on the tumor growth of tumor-bearing mice. In addition, the changing pattern of the splenic lymphocyte subsets of tumor-bearing mice was evaluated by flow cytometry. OK-432 at a dose of 0.1, 1 or 10 KE was administered orally every 3 days or every other day for 30 days to subcutaneously Meth A tumor-inoculated mice. The tumor growth was significantly inhibited in the 1 KE every 3 days group, in the 1 KE every other day group and in the 10 KE every 3 days group. In the 10 KE every other day group, OK-432 inhibited the tumor growth on days 10 and 20, while the agent did not show a marked inhibitory effect on day 30. The percentages of splenic L3T4-positive cells and splenic asialo GM1-positive cells were significantly increased in the 1 KE every other day group, while the Lyt2+/Thy1.2+ ratio was decreased. On the other hand, in the 10 KE every other day group, OK-432 showed no effect on the percentages of splenic L3T4-positive cells and Lyt2+/Thy1.2+ ratio on days 20 and 30. Our results suggest that the antitumor effect of oral administration of OK-432 may be correlated with the changing pattern of L3T4-positive cells and Lyt2+/Thy1.2+ ratio.