Induction and clinical utilization of lymphokine-activated killer cells in patients with gastrointestinal tract cancers

In 18 patients with cancers of the gastrointestinal tract, lymphokine-activated killer (LAK) cell activity was studied and compared with that of healthy subjects. After cultivation with 10(3) iu/mL of recombinant interleukin-2, the cytotoxicity of patients' lymphoid cells was increased from 13.6 +/- 6.8% to 76.2 +/- 19.5% against Daudi cells and from 12.8 +/- 8.1% to 76.2 +/- 19.5% against K-562 cells. Based on these results, autologous LAK cells were given to patients. LAK cells injected into subdermal metastatic tumours demonstrated a significant inhibitory effect on tumour growth in comparison with that of control tumour nodules. Of four patients with metastatic tumours in the liver, to whom LAK cells were administered via the hepatic artery, tumour size was reduced by about 25% (minor response) in one patient, with a decrease of computerized tomography attenuation in the tumours occurring in the other three patients.     

Defective immunological functions associated with abnormal lymphokine-activated killer activity in patients with hepatocellular carcinoma

The in vitro lymphokine-activated killer (LAK) activity of peripheral blood mononuclear cells (PBMC) from 36 patients with hepatocellular carcinoma was investigated. The activity was greatly diminished in 13 patients and enhanced in seven patients. A flow cytometric study showed that the percentage of OKM1+, Leu-7+-11b+, and Leu-7-11b+ fractions in PBMC was decreased and the percentage of OKT8+ and Leu7+11- fractions was increased significantly in the 13 patients with lower LAK activity, compared with the values of the seven higher LAK activity patients. Furthermore, the response of PBMC to interleukin-2 (IL-2) was deficient in the lower activity group. However, there was no significant difference in IL-2 production by PBMC, IL-2 receptor (p55) expression of PBMC and mitogen (Con-A, PHA) response of PBMC between the two groups. These findings indicate the possibility that diminished LAK activity in hepatoma patients is due to a decreased number of LAK precursor cells and a defective response of LAK precursor cells to IL-2.     

Induction of lymphokine-activated killer cells against human leukemia cells in vitro

Summary The induction of lymphokine-activated killer (LAK) cells against fresh human leukemia cells was investigated. Two thirds of the 62 leukemias examined were susceptible to the lytic effect of allogeneic IL-2 induced LAK cells in vitro. No substantial differences could be detected between myeloid or lymphoid leukemias or with regard to the FAB subtype or the immunophenotype. Culturing mononuclear cells from peripheral blood or bonemarrow of leukemia patients with IL-2 resulted in an expansion of residual large granular lymphocytes and development of cytotoxic activity. The combination of IL-2 with IFN-gamma or the presence of tumor cells during the activation process led to an enhancement of LAK cell cytotoxicity. These results suggest that LAK cells may be useful in the treatment of leukemia.Supported by the Deutsche Krebshilfe e.V./Mildred Scheel Stiftung für Krebsforschung, Bonn (W 19/87/Te 1)     

LAK／IL－2不同给药途径对肝癌治疗效果的比较研究

应用ＬＡＫ／ＩＬ－２单独或与经肝动脉插管化疗药物灌注或栓塞治疗（ＴＡＣＥ）结合，分别通过肿瘤供血动脉、外周静脉或浆膜腔输注等多种途径，治疗肝癌１６例。Ｂ组、Ｃ组均为Ⅱ期与Ⅲ期病例，结合ＴＡＣＥ治疗有效率达６４．３％（９／１４）；而Ａ组２例Ｉ期小肝癌病例，单纯予以肝动脉插管输注ＬＡＫ／ＩＬ－２等治疗无效。提示这种过继免疫治疗不宜单独使用，而在ＴＡＣＥ术后患者免疫功能开始恢复阶段，作为综合治疗的一项措施是十分有益的。     

Treatment of non-resectable hepatocellular carcinoma with autologous tumor-pulsed dendritic cells

BACKGROUND: The response of hepatocellular carcinoma (HCC) to therapy is often disappointing and new modalities of treatment are clearly needed. Active immunotherapy based on the injection of autologous dendritic cells (DC) co-cultured ex vivo with tumor antigens has been used in pilot studies in various malignancies such as melanoma and lymphoma with encouraging results. METHODS: In the present paper, the preparation and exposure of patient DC to autologous HCC antigens and re-injection in an attempt to elicit antitumor immune responses are described. RESULTS: Therapy was given to two patients, one with hepatitis C and one with hepatitis B, who had large, multiple HCC and for whom no other therapy was available. No significant side-effects were observed. The clinical course was unchanged in one patient, who died a few months later. The other patient, whose initial prognosis was considered poor, is still alive and well more than 3 years later with evidence of slowing of tumor growth based on organ imaging. CONCLUSIONS: It is concluded that HCC may be a malignancy worthy of DC trials and sufficient details in the present paper are given for the protocol to be copied or modified.     

Immunotherapy of hepatocellular carcinoma with autologous lymphokine-activated killer cells and/or recombinant interleukin-2

Summary Five patients with hepatocellular carcinoma were subjected to immunotherapy: three patients were treated by adoptive immunotherapy with lymphokine-activated killer (LAK) cells and recombinant interleukin-2 (rIL-2), and two patients by systemic administration of rIL-2 alone. In one patient with diffuse-type hepatocellular carcinoma and portal vein thrombosis who was treated by infusion of LAK cells (a total number of 1.5x10¹⁰ cells/13 doses) and continuous rIL-2 administration (a total dose of 1.25x10⁸ units) via a percultaneously placed hepatic arterial catheter, the size of the tumor reduced dramatically and the portal vein thrombosis retracted. In two patients who had LAK cells infused (totals of 6.6x10⁹ cells/4 doses and 3.1x10⁹ cells/2 doses, respectively) during hepatic angiogram followed by systemic administration of rIL-2 twice a day, no clinical improvement was noticed. In two patients who received rIL-2 alone systemically (total doses of 8.9x10⁷ and 5.5x10⁷ units, respectively), neither clinical improvement nor severe side effects were observed. The results suggest that adoptive immunotherapy combined with continuous local administration of rIL-2 via a percutaneously placed hepatic arterial catheter may be an effective therapy without apparent side effects for patients with hepatocellular carcinoma who cannot be treated by conventional cancer therapy.     

The management of hepatocellular carcinoma around the world: a comparison of guidelines from 2001 to 2011

In the past 10 years, many guidelines for hepatocellular carcinoma (HCC) have been published worldwide. To promote standard care for HCC, we systematically evaluated 17 current guidelines for HCC around the world, including 5 guidelines from the USA, 7 from Asia and 5 from Europe, according to the selection criteria of credibility influence and multi-faceted. After a systematic evaluation, we found that these guidelines have both similarities and differences in terms of what organizations or bodies drafted the guidelines and the approach, applicability, content and recent updates of the guidelines as well as in terms of diagnostic and treatment algorithms. The differences could be attributed to various aetiological factors, high-risk patients, health systems, health resources, medical technology, treatment choices and income levels in different countries. Besides, although the full implementation of guidelines could benefit clinicians, patients and authorities, there is still a gap between projected goals and implementation. The factors potentially influencing implementation are what organizations or bodies are drafting guidelines, content and emphasis, modification and consistency of guidelines. Comparative analysis suggested that countries pay close attention to targeted audiences, a basis in evidence, a basis in available resources, applicable patients and systematic evaluation when establishing and implementing domestic guidelines for HCC.     

Preventive antitumor activity against hepatocellular carcinoma (HCC) induced by immunization with fusions of dendritic cells and HCC cells in mice

BACKGROUND: The prevention of recurrence of hepatocellular carcinoma (HCC) after treatment is very important for improvement of the prognosis of HCC patients. Dendritic cells (DCs) are potent antigen-presenting cells that can prime naive T cells to induce a primary immune response. We attempted to induce preventive antitumor immunity against HCC by immunizing BALB/c mice with fusions of DCs and HCC cells. METHODS: Murine bone marrow-derived DCs and a murine HCC cell line. BNL cells, were fused by treatment with 50% polyethyleneglvcol (PEG). Fusion efficacy was assessed by the analysis of fusions of BNL cells stained with red fluorescent dye and DCs stained with green fluorescent dye. Mice injected intravenously with DC/BNL fusions were challenged by BNL cell inoculation. RESULTS: About 30% of the PEG-treated non-adherent cells with both fluorescences were considered to be fusion cells. The cell fraction of DC/BNL fusions showed phenotypes of DCs, MHC class II, CD80, CD86, and intercellular adhesion molecule (ICAM)-1, which were not expressed on BNL cells. Mice immunized with the fusions were protected against the inoculation of BNL tumor cells, whereas injection with a mixture of DCs and BNL cells not treated with PEG did not provide significant resistance against BNL cell inoculation. Splenocytes from DC/BNL fusion-immunized mice showed lytic activity against BNL cells. CONCLUSIONS: These results demonstrate that immunization with fusions of DCs and HCC cells is capable of inducing preventive antitumor immunity against HCC.     

Evidence of aberrant lipid metabolism in hepatitis C and hepatocellular carcinoma

Objectives

Lipids are linked to many pathological processes including hepatic steatosis and liver malignancy. This study aimed to explore lipid metabolism in hepatitis C virus (HCV) and HCV-related hepatocellular carcinoma (HCC).

Methods

Serum lipids were measured in normal, HCV and HCV-HCC patients. Whole-genome microarray was performed to identify potential signature genes involved in lipid metabolism characterizing normal vs. HCV vs. HCV-HCC conditions.

Results

Serum cholesterol was significantly reduced in HCV and HCV-HCC patients compared with normal controls, whereas there was no difference in glucose and triglycerides. Microarray analysis identified 224 probe sets with known functional roles in lipid metabolism (anova, 1.5-fold, P ≤ 0.001). Gene-mediated fatty acid (FA) de novo synthesis and uptake were upregulated in HCV and this upregulation was further enhanced in HCC. Genes involved in FA oxidation were downregulated in both the HCV and HCC groups. The abnormality of cholesterol metabolism in HCV was associated with downregulation of genes involved in cholesterol biosynthesis, absorption and transportation and bile acid synthesis; this abnormality was further intensified in HCC.

Conclusions

Our data support the notion that HCV-related lipid metabolic abnormalities may contribute to hepatic steatosis and the development of cancer. Identification of these aberrations would stratify patients and improve treatment algorithms.     

Systemic administration of liposome-encapsulated OK-432 prolongs the survival of rats with hepatocellular carcinoma through the induction of IFN-gamma-producing hepatic lymphocytes

BACKGROUND: OK-432 is known to increase the host antitumor response. We previously reported that systemic administration of OK-432 (OK-Lipo) specifically induced hepatic lymphocytes in mice. Here we aimed to investigate the antitumor effect of OK-Lipo on hepatocellular carcinomas (HCC) in experimental rats. METHODS: Diethylnitrosamine was administered for 12 weeks to all rats (n = 36). Rats were divided into three groups of 12 rats each. One group was injected with OK-Lipo from week 5 (OK-5w group) and another from week 9 (OK-9w group). A control group was injected with saline from week 5 (Non-OK group). At week 13, five rats from each group were used for histological analysis and immunofluorescence assays (surface phenotypic and intracellular cytokine analysis of the mononuclear cells in the liver, spleen and peripheral blood). The remaining rats were observed for the remainder of their survival period. RESULTS: The mean survival times of Non-OK, OK-5w, and OK-9w groups differed significantly (98.0 +/- 5.3 days, 116.0 +/- 5.8 days, and 106.0 +/- 5.4 days, respectively, P < 0.01). Histological examination revealed many apoptotic tumor cells, infiltration of lymphocytes and macrophages in the OK-5w group. The two-color immunofluorescence assay showed that the proportion of natural killer (NK) cells and IFN-gamma-producing cells in the liver were significantly higher in the OK-5w group. CONCLUSIONS: These findings showed that systemic administration of OK-Lipo contributed to prolonging the survival of rats with HCC. OK-Lipo induced NK cells and IFN-gamma-producing cells specifically in the liver and these cells seemed to reduce hepatocarcinogenesis and tumor growth.     

肝细胞癌分期系统的临床意义与进展

大多数实体瘤患者肿瘤的诊断和分期与其生存期有关,且直接影响治疗指征。肝细胞癌患者生存期的预测较复杂,目前国际上对肝细胞癌分期系统尚无统一的认识。若分期系统只考虑预后参数,如肿瘤、淋巴结、转移或Child—Pugh分级中的一种,那该分期系统是无用的。已建立的几个分期系统,均对终末期患者有确定能力而与治疗无关联。巴塞罗纳临床肝细胞癌分期系统是在几个队列和临床随机研究结果基础之上建立的,集肿瘤状态、治疗方案与预测生存期为一体的分期系统。     

Tumor-detecting capacity and clinical usefulness of SPIO-MRI in patients with hepatocellular carcinoma

The tumor-detecting capacity and clinical usefulness of superparamagnetic iron oxide (SPIO) magnetic resonance imaging (MRI) were examined in patients with hepatocellular carcinoma. The tumor detection rate of SPIO-MRI (64.5%) was comparable to those of dynamic computed tomography (CT) and plain MRI, but lower than that for Gd dynamic MRI (93.5%; P < 0.01%). A combination of Gd dynamic MRI and SPIO-MRI improved the detection rate; further, the tumor stage with respect to tumor blood-flow pattern was predicted by combining plain MRI with SPIO-MRI. This combination procedure may also be useful for selecting therapeutic strategies. Received: November 26, 1999 / Accepted: June 23, 2000     

葛根素对人肝癌细胞HepG2侵袭与迁移能力的影响

目的探索葛根素对人肝癌细胞HepG2侵袭与迁移能力的影响及其相关分子机制。方法将人肝癌细胞HepG2分为两组,实验组用60μg/ml、30μg/ml的葛根素进行培养,对照组用加入等体积的葛根素溶解剂二甲基亚砜(DMSO)的培养基培养;利用细胞划痕实验检测两组细胞的迁移能力;利用Transwell小室检测两组细胞的侵袭能力;通过蛋白免疫印迹检测波形蛋白(vimentin)、E-钙黏蛋白(E-cadherin)、基质金属蛋白酶2/9(matrix metalloproteinase2/9,MMP2/9)。结果人肝癌细胞HepG2在经过葛根素培养后,细胞划痕的愈合能力下降,Transwell小室细胞的侵袭能力下降;E-cadherin表达上升(P<0.05),vimentin、MMP2和MMP9的表达下调(P<0.05)。结论葛根素通过上皮细胞间质转化(EMT)途径导致人肝癌细胞HepG2侵袭转移能力下降。     

Combined hepatocellular carcinoma and cholangiocarcinoma growing into the common bile duct

Received: August 7, 2000 / Accepted: December 22, 2000     

Percutaneous ethanol injection for the treatment of small hepatocellular carcinoma. Study of 95 patients

Percutaneous ethanol injection (PEI) was applied to 120 lesions in 95 patients with hepatocellular carcinomas (HCC) smaller than 3 cm in the past 6 years. All main target tumours, in 67 patients who had been followed by sonography for more than 6 months after PEI, decreased in size; 28 tumours (41.8%) became undetectable and have remained so until now. The 1-, 2-, 3-, 4- and 5-year survival rates calculated by the Kaplan-Meier method were 93%, 81%, 65%, 52% and 28% respectively. These survival rates were better than those of patients with HCC smaller than 3 cm who did not receive anticancer treatment (P less than 0.01). The survival of patients of the Child's A or Child's B status was better than that of those with Child's C disease. Recurrence occurred in areas within the liver different from the original lesion in 34% in one year, 61% in two years and 66% in three years after PEI. PEI was then repeated in 61% of such patients.     

Patient-derived dendritic cells transduced with an a-fetoprotein-encoding adenovirus and co-cultured with autologous cytokine-induced lymphocytes induce a specific and strong immune response against hepatocellular carcinoma cells.

BACKGROUND/AIMS: Breaking immunologic tolerance towards the hepatocellular carcinoma (HCC)-associated alpha-fetoprotein (AFP) antigen is possible. The use of this potential for the treatment of immunocompromised HCC patients is limited. In this study, we analyzed whether dendritic cells (DCs) from HCC patients transduced with a human AFP (hAFP)-expressing adenovirus and co-cultured with cytokine-induced killer (CIK) cells can induce a strong specific immune response against HCC-cells. METHODS: An hAFP-encoding adenovirus (Ad-hAFP) was generated. DCs from healthy donors or patients were transduced at a very high efficacy. Afterwards, DCs were co-cultured with autologous CIK-cells, and their ability to lyse HCC-cells was analyzed. RESULTS: AFP-transduced DCs stimulated CIK cells strongly to lyse about 70% of AFP-expressing HCC cells. Cytotoxicity was significantly higher when lymphocytes were co-cultured with Ad-hAFP-transduced DCs than with Ad-mock-transduced DCs, indicating an AFP-specific immune response. More interestingly, CIK cells from patients with AFP-positive HCC could be stimulated to lyse AFP-expressing HCC cells as effectively as CIK cells from healthy individuals and stronger than CIK cells from patients without AFP-expressing HCC. CONCLUSIONS: The data demonstrate that patient-derived DCs that were transduced with an AFP-expressing adenovirus and co-cultured with autologous CIK cells induce an AFP-specific, strong immune response against HCC cells. Therefore, this approach may have a potential for an adoptive and/or DC-based immunotherapy for HCC patients.     

肝癌细胞的诱导分化及凋亡的研究

本文把肝癌细胞（SMMC-7721）作为靶细胞采用TRAP银染法、Westernblot法、电镜等方法,分别检测对肝癌细胞的全反式维甲酸（ATRA）的诱导凋亡及奥曲肽的诱导分化作用。ATRA在TRAP银染中未见梯状条带,然而在Westernblot分别于32KD、20KD处显示较强的Caspase-3的表达。提示,ATRA有较强的诱导凋亡的作用。在奥曲肽的刺激下可见肝细胞核浓缩、微绒毛减少、线粒体空泡化等变化,而且促进Caspase-3的表达并使其激活,从而诱导肝癌细胞的分化及凋亡。结果显示,上述两种制剂分别对肝癌细胞具有较强的诱导分化及促进凋亡的作用。     

The development of hepatocellular carcinoma from liver cirrhosis during a follow-up study

A prospective study was carried out in 126 cases with liver cirrhosis attending the outpatient clinic of Hepatology of the Department of Internal Medicine, Dr. Cipto Mangunkusmo Hospital Jakarta, between August 1,1982 and Dec. 31, 1985. The patients consisted of 82 men and 44 women and there were 45 HBsAg positive cases (36.7%). HBeAg was posotive in 35.6% (16/45) and 40.0% were antiHBe positive while both markers were negative in 24.4%. During the follow-up study 27 cases died. The cause of death was due to variceal bleeding in 9 cases (33.3%) and hepatic failure in 9 cases (33.3%). In 6 cases (22.2%) hepatocellular carcinoma (HCC) was the cause of death. Nine cases out of 94 traceable cases (13.7%) were developed HCC. They consisted of 4 cases out of 29 cases (13.8%) with HBsAg positive and 5 cases out of 65 cases (7.7% ). But no significant difference was observed between both groups. The length of observation period from the first time of diagnosis until development of HCC was from 1 to 6 years with an average of 2.9 years. Financial support from Japan Society for Promotion Science is acknowledged.