卵巢癌患者手术前后IL-2、IL-6及T细胞亚群水平改变的临床意义

目的 探讨卵巢癌患者细胞免疫状况及IL-2、IL-6及T细胞亚群与临床的关系。方法ELISA法检测血清IL-2、IL-6水平，免疫荧光法检测T细胞亚群。结果40例卵巢恶性肿瘤患者手术前及术后7天IL-2产生能力降低，IL-6水平升高，与术后第6疗程化疗结束后4周相比差异有显著意义(P＜0．01)；术前临床Ⅲ、Ⅳ期患者IL-2活性低于Ⅰ、Ⅱ期患者，IL-6水平明显高于Ⅰ、Ⅱ期水平(P＜0．05)；CD4细胞显著减少，CD4／CD8明显降低，与Ⅰ、Ⅱ期患者比较差异有显著意义(P＜0．01)；术后复发的6例患者血清IL-6持续增高。结论 对IL-2、IL-6及T细胞亚群的定期检测有助于观察卵巢恶性肿瘤患者病情变化，为其预后判断提供重要的参考。     

IL-12抗肿瘤基因治疗的研究进展

IL-12具有强大的抗瘤活性，在肿瘤生物治疗中占有重要地位。近年来IL-12基因疗法成为研究的热点，各国学者采用不同的治疗途径及基因联合方式进行了大量尝试，以求其最佳抗瘤效果，现就此作一综述。     

卵巢癌患者手术前后IL-2、IL-6及T细胞亚群水平改变的临床意义

目的探讨卵巢癌患者细胞免疫状况及IL-2、IL-6及T细胞亚群与临床的关系.方法 ELISA法检测血清IL-2、IL-6水平,免疫荧光法检测T细胞亚群.结果 40例卵巢恶性肿瘤患者手术前及术后7天IL-2产生能力降低,IL-6水平升高,与术后第6疗程化疗结束后4周相比差异有显著意义(P＜0.01);术前临床Ⅲ、Ⅳ期患者IL-2活性低于Ⅰ、Ⅱ期患者,IL-6水平明显高于Ⅰ、Ⅱ期水平(P＜0.05);CD4细胞显著减少,CD4/CD8明显降低,与Ⅰ、Ⅱ期患者比较差异有显著意义(P＜0.01);术后复发的6例患者血清IL-6持续增高.结论对IL-2、IL-6及T细胞亚群的定期检测有助于观察卵巢恶性肿瘤患者病情变化,为其预后判断提供重要的参考.     

联合应用IL-2和IL-7对T细胞体外增殖及功能的增强效应

本文研究了联合应用IL-2和IL-7在体外T细胞培养系统中对小鼠T细胞生长及功能的调节作用。结果表明:单独使用IL-2或IL-7均能增强T细胞对ConA或特异性抗原刺激诱导的增殖反应,但联合使用IL-2和IL-7具有更加显著的协同刺激作用。C57BL/6小鼠抗FBL-3肿瘤特异性T细胞系在IL-2与IL-7联合培养系统中,不但T细胞数量明显增加,且能在保持特异性功能的同时,延长T细胞体外抗原刺激周期。此外,这一细胞因子的组合方案,可大大降低IL-2的应用剂量,避免大剂量IL-2临床应用时所产生的副作用。     

IL-12抗肿瘤基因治疗的研究进展

IL-12具有强大的抗瘤活性,在肿瘤生物治疗中占有重要地位.近年来IL-12基因疗法成为研究的热点,各国学者采用不同的治疗途径及基因联合方式进行了大量尝试,以求其最佳抗瘤效果,现就此作一综述.     

IL-15对树突状细胞激活的T淋巴细胞免疫反应的作用观察

  目的  观察IL-15和IL-2对树突状细胞疫苗(dendritic cells, DC)激活的淋巴细胞免疫反应的作用。  方法  取健康小鼠的脾淋巴细胞, 在体外采用肿瘤抗原负载的DC疫苗与脾淋巴细胞混合后, 分别加入IL-15或IL-2培养7d。采用流式细胞术检测淋巴细胞免疫表型的变化; 使用酶联免疫斑点实验(ELISPOT)检测分泌干扰素(interferon, IFN)-γ的细胞数量; 利用51Cr释放法检测淋巴细胞对肿瘤细胞的杀伤活性。  结果  IL-15联合DC疫苗增强了T细胞反应, 包括淋巴细胞免疫表型和功能的改变。  结论  与IL-2相比, IL-15能更好的增强DC疫苗诱发的免疫反应, 为IL-15与DC疫苗的联合应用提供了一定的依据。      

IL-2基因修饰的细胞毒 T淋巴细胞抗肿瘤效应的研究

目的：了解IL－2基因修饰的细胞毒T淋巴细胞（cytotoxic T lymphocytes,CTL)的增殖和杀伤活性,探索细胞因子基因疗法及被动免疫治疗脑胶质瘤的新途径。方法：用昆明种小鼠的脾细胞体外诱导CTL,用腺病毒载体转染IL－2基因,观察其体外增殖活性和样伤活性,再用G422小鼠胶质母细胞瘤细胞建立肺转移瘤模型,36h后经过继回输,2周后计数肺的肿瘤结节,观察IL－2基因转染的CTL对实验性肺转移瘤的治疗作用。结果：重组腺病毒载体在MOI（multipliciy of infection)为100时,转染率达96．8％,IL－2基因修饰CTL增殖活性、IL－2的分泌（248u)和体外杀伤活性（36．4％）明显增强,对实验性肺转移瘤的治疗作用都显著增强,肺转移结节数（28）显著减少（P＜0．01）。结论：IL－2基因转染的CTL过继回输,可直接杀伤和诱导激活机体抗肿瘤免疫反应,使体内抗肿瘤效果显著增强,有效抑制实验性肺转移瘤的生长,为胶质瘤的过继免疫治疗提供了新的思路和实验依据。     

胃癌患者IL-10,IL-12的检测及其临床意义

目的检测胃癌患者IL-10、IL-12在癌组织和癌周组织中的含量及术前和术后1周、2周血清中浓度的变化并探讨其意义。方法用ELISA法检测了22例胃癌患者IL-10、IL-12在癌组织和癌周组织中的含量及术前和术后1周、2周血清中的浓度,以16例健康人为对照。结果胃癌患者IL-10含量在癌组织中高于癌周组织,其中IL-10含量在Ⅲ、Ⅳ期癌组织和癌周组织中均高于Ⅰ、Ⅱ期癌组织和癌周组织,IL-12含量则相反。胃癌患者血清中IL-10浓度升高,且血清IL-10浓度Ⅲ、Ⅳ期较Ⅰ、Ⅱ期高,术后1周、2周血清中IL-10浓度逐渐下降,但仍高于对照组浓度,IL-12浓度则相反。结论胃癌细胞可能分泌IL-1 0抑制IL-1 2,使机体的免疫力降低。IL-10,IL-12的检测可反映胃癌病期、判断疗效和估计预后。     

IL-18促进肺癌细胞转移的功能鉴定

目的 探讨IL-18是否具有促进肺癌细胞转移的功能以及通过何种途径发挥作用。方法 以人肺巨细胞癌高、低转移株为肿瘤转移的研究模型,采用Western blot和半定量RT-PCR结合Northern blot法检测IL-18蛋白或mRNA在肺臣细胞癌高、低转移株的差异表达;随后采用基因转染技术将IL-18的正义或反义表达质粒分别转染肺巨细胞癌低或高转移株,比较IL-18稳定转染细胞转移表型的改变。采用：MICS系统检测稳定转染细胞体外迁移和侵袭能力的改变,采用Western blot法检测稳定转染细胞转移表型相关指标的改变,从而确定IL-18通过何种途径促进肺癌细胞转移。结果 由于IL-18蛋白和mRNA仅在肺巨细胞癌高转移株检测到,因此,将IL-18正义表达质粒转染肺巨细胞癌低转移株后,可检测到IL-18融合蛋白的表达,且稳定转染细胞出现了细胞体外迁移能力的显著上调和上皮细胞间黏附分子E-cadherin蛋白的显著下调;相反,将IL-18反义表达质粒转染肺巨细胞癌高转移株后,可检测到内源性IL-18蛋白和mRNA水平的下调,且稳定转染细胞出现了细胞体外侵袭能力的显著下调和E-cadherin蛋白的显著上调。结论 IL-18可能通过抑制上皮细胞问黏附分子E-cadherin蛋白的表达来促进肺癌细胞的转移。     

香菇多糖抑制乳腺癌4T1细胞小鼠移植瘤增殖机制研究

目的 观察香菇多糖对乳腺癌小鼠的脾脏、肿瘤组织及外周血中白细胞介素-35(IL-35)表达影响,探讨香菇多糖抑制乳腺细胞癌的可能作用机制.方法 30只雌性Balb/c小鼠分为对照组、模型组和香菇多糖给药组(200 mg/kg),每组10只.注射4T1细胞建立4T1乳腺癌小鼠模型,给药组持续灌服给药香菇多糖,每只小鼠给药...     

Blocking hypoxia-induced autophagy in tumors restores cytotoxic T-cell activity and promotes regression

The relationship between hypoxic stress, autophagy, and specific cell-mediated cytotoxicity remains unknown. This study shows that hypoxia-induced resistance of lung tumor to cytolytic T lymphocyte (CTL)-mediated lysis is associated with autophagy induction in target cells. In turn, this correlates with STAT3 phosphorylation on tyrosine 705 residue (pSTAT3) and HIF-1α accumulation. Inhibition of autophagy by siRNA targeting of either beclin1 or Atg5 resulted in impairment of pSTAT3 and restoration of hypoxic tumor cell susceptibility to CTL-mediated lysis. Furthermore, inhibition of pSTAT3 in hypoxic Atg5 or beclin1-targeted tumor cells was found to be associated with the inhibition Src kinase (pSrc). Autophagy-induced pSTAT3 and pSrc regulation seemed to involve the ubiquitin proteasome system and p62/SQSTM1. In vivo experiments using B16-F10 melanoma tumor cells indicated that depletion of beclin1 resulted in an inhibition of B16-F10 tumor growth and increased tumor apoptosis. Moreover, in vivo inhibition of autophagy by hydroxychloroquine in B16-F10 tumor-bearing mice and mice vaccinated with tyrosinase-related protein-2 peptide dramatically increased tumor growth inhibition. Collectively, this study establishes a novel functional link between hypoxia-induced autophagy and the regulation of antigen-specific T-cell lysis and points to a major role of autophagy in the control of in vivo tumor growth.     

Interleukin-12 requires initial CD80-mediated T-cell activation to support immune responses toward human breast and ovarian carcinoma.

One possible reason for the poor immunogenicity of tumors is the induction of peripheral tolerance by tumor cells that fail to deliver costimulatory signals. Furthermore, T cells stimulated with wild-type tumor cells often fail to secrete cytokines. The present study has been undertaken to identify cytokines that cooperate with CD80 in T-cell activation in vitro toward human breast and ovarian carcinoma cell lines. Tumor cell-mediated T-lymphocyte activation was analyzed directly in allogeneic mixed lymphocyte/tumor cell cultures as proliferation and effector functions were assessed in cytotoxic T-cell assays. Interleukin-7 (IL-7) amplified the proliferative response toward CD80-transfected breast and ovarian carcinomas and stimulated predominantly CD4+ T lymphocytes. IL-12 represses the proliferative response of naive T cells but cooperates with CD80-mediated activation during secondary stimulations. In long-term T-cell cultures, IL-12 synergizes with CD80 expression to stimulate cytolytic CD8+ T-cell lines, which recognize a breast carcinoma line in a human histocompatibility leukocyte antigen-restricted manner. These studies illustrate that costimulation is necessary for tumor cells to function as alloantigen-presenting cells. Furthermore, when added after the priming of T cells with CD80-transfected tumor cells, IL-12 could be helpful in propagating sufficient T-cell numbers to be used in adoptive transfers during cellular immunotherapy.     

Restoration of T-cell function and induction of antitumor immune response in T-cell-depressed spontaneously hypertensive rats by treatment with thymosin fraction 5

We studied the effect of treatment with Thymosin Fraction 5 (Fr-5) on the restoration of T-cell functions and the induction of antitumor immunity in spontaneously hypertensive rats (SHR) with congenital T-cell depression. SHR showed a reduced number of rosette-forming thymocytes early in their lives, and in vitro incubation of SHR thymocytes with Fr-5 restored the numbers of rosette-forming T-cells to the normal level for Wistar/HMK rats, the original strain of SHR. In vivo treatment of SHR with various doses of Fr-5 (0.5, 1.0, or 2.0 mg/kg, 6 times every other day) also increased significantly the blastogenic responses of their spleen cells to phytohemagglutinin but failed to promote plaque-forming cell responses to sheep red blood cells. These immunological restorative effects by Fr-5 were dose dependent. In contrast, treatment with lower doses of Fr-5 (0.25 or 0.50 mg/kg) showed greater curative effects on a high antigenic fibrosarcoma (SMT-6) than did treatment with higher doses of Fr-5 (1.0 or 2.0 mg/kg). This was confirmed by the fact that treatment with a 0.5-mg/kg dose of Fr-5 caused a significant suppressive effect on the growth of a weakly antigenic and highly metastatic adenocarcinoma (SST-2) in SHR with a consequent prolongation of survival days, whereas treatment with a 1.0- or 2.0-mg/kg dose of Fr-5 was without any effect. In order to clarify this mechanism, we studied the effect of pretreatment with cyclophosphamide (CY) on the development of antitumor delayed-type hypersensitivity (DTH) reaction in the SMT-6-bearing SHR treated with Fr-5 (0.5 or 2.0 mg/kg). Treatment with 0.5 mg of Fr-5 per kg significantly increased the DTH reaction to SMT-6 cells in both CY-pretreated and untreated SHR. In contrast, treatment with 2.0 mg of Fr-5 per kg produced a significant antitumor DTH reaction in SHR pretreated with CY but failed to induce the DTH reaction in SHR untreated with CY. These results suggest that higher doses of Fr-5 may induce preferentially suppressor T-cells rather than killer T-cells in tumor-bearing SHR with congenital T-cell depression.     

The effect of 5-fluorouracil on rat adrenal function

This study was undertaken to determine the effects of 5-fluorouracil on corticosteroidogenesis and adrenal size in the rat. The rats were injected intraperitoneally with 5-FU (12.5 mg kg-1 or 25 mg kg-1), normal saline or actinomycin D (0.02 mg kg-1), daily for 10 days. High-dose 5-FU induces adrenal hyperplasia, in association with mild impairment of corticosteroidogenesis (manifest by lower corticosterone and higher ACTH levels). The response to low-dose 5-FU and actinomycin D is of lesser adrenal hyperplasia, relative to high-dose 5-FU (P less than 0.05) and elevated corticosterone levels. There may be a dose-related effect on the suppression of corticosteroidogenesis in the rat by 5-FU.     

Interleukin-12 gene therapy vaccines: directing the immune system against minimal residual leukemia

Current overall survival rates for patients with AML remain poor and there is need for novel therapeutic approaches. One such approach is to use the patient's own immune system to eliminate minimal residual disease. Recent advances in genetic manipulation of tumor cells, together with a better understanding of the immune mechanisms controlling the host-tumor relationship have led to a flurry of preclinical and clinical studies on tumor cell vaccines. Here we present a brief overview of genetic manipulation of tumor cells, and highlight important principles of cancer immunity and cancer vaccines. Special emphasis is given on recent work on the role of interleukin-12 (IL-12) based vaccines in murine AML. These studies have shown that vaccines with AML cells, genetically modified to secrete IL-12, are potent stimulators of the immune system and lead to the development of both prophylactic and therapeutic anti-leukemia immunity.     

Interleukin 6 perfusion stimulates reconstitution of the immune and hematopoietic systems after 5-fluorouracil treatment

Effects of interleukin 6 (IL-6) on the functional capacity of the immune and hematopoietic systems in 5-fluorouracil (5-FU)-treated mice were determined. IL-6 (5 x 10(4) units/mouse/day) was administered s.c. for 7 days by implantation of an osmotic pump, since it was demonstrated that a much higher increase in the primary response to sheep RBC was observed by administration of slowly released rather than daily s.c. injection of IL-6. IL-6 perfusion significantly augmented anti-sheep RBC antibody responses depressed by 5-FU (150 mg/kg) treatment. IL-6 also was shown to stimulate hematological recovery in mice treated with 5-FU. Namely, IL-6 perfusion accelerated the recovery of the number of hematopoietic stem cells, granulocyte-macrophage progenitors, and mature neutrophils in the spleen, although IL-6 did not stimulate the recovery of the neutrophil count in blood. Recovery of the platelet count in blood was stimulated by IL-6. Furthermore, it was found that the endogenous IL-6 level in serum increased after 5-FU treatment, which suggests that IL-6 may play some role in the recovery of the immune and hematopoietic systems. Finally, we examined the effect of IL-6 on the survival of mice treated with a higher dosage of 5-FU (300 mg/kg). IL-6 perfusion produced a distinct increase in survival rate at Day 30 (74% versus 28%). It is of note that the number of bacteria (identified as Escherichia coli) cultured from the spleen and the liver decreased in IL-6-perfused mice. This IL-6-induced effect was accompanied by enhancement of an oxidative burst response. Moreover, the anti-E. coli antibody titer in serum was higher in IL-6-perfused mice than in control mice. These results suggest the possible use of IL-6 for stimulating the reconstitution of the immune and hematopoietic systems after chemotherapy treatment.     

The effect of 5-fluorouracil and alpha interferon and 5-fluorouracil and leucovorin on cellular anti-tumour immune responses in patients with advanced colorectal cancer.

Interferon alpha (IFN-alpha) enhances the activity of 5-fluorouracil (5-FU) in the treatment of advanced colorectal cancer although the mechanism is not understood. We have investigated the effect of this combination on cellular immunity and compared this with standard therapy of 5-FU/L-leucovorin, in 24 patients with advanced colorectal cancer. This study has demonstrated an enhancement of the cellular immune response in patients given 5-FU/IFN-alpha with augmentation of natural killer (NK) cell function and abrogation of 5-FU-induced suppression of lymphokine-activated killer (LAK) cell activity.