picibanil and Carcinoma--Hepatocellular

The current curative treatments for hepatocellular carcinoma (HCC) do not effectively prevent tumor recurrence. Dendritic cell (DC)-based immunotherapy can be a novel strategy targeting tumor recurrence. Here, we evaluated the bioactivity and beneficial effects of DC infusion of HCC tissues following transcatheter hepatic arterial embolization (TAE). 5x10(6) of monocyte- derived DCs were administered through an arterial catheter during TAE treatment procedures in patients with hepatitis C virus-related cirrhosis and HCC. In DC preparation, peptide-stimulated DCs [HLA-DR(+)CD86(+)CD14(-)] were phenotypically immature [CD80(low)CD83(low)], while OK-432-stimulated DCs highly expressed the activation marker CD83. Following the transfer, there was no clinical or serological evidence of adverse events in any patients in addition to those due to TAE. Most interestingly, survival analysis indicated that the patients treated with OK-432-stimulated DCs prolonged the recurrence-free survivals when compared with the historical controls treated with TAE alone. Collectively, OK- 432-stimulated DC transfer to HCC tissues following TAE treatment contributes to the induction of beneficial antitumor immune responses and the prolongation of recurrence-free survivals, providing a plausible strategy to reduce the tumor recurrence rates of HCC.

Topics: Antineoplastic Agents; Carcinoma, Hepatocellular; Dendritic Cells; Humans; Immunotherapy; Liver Neoplasms; Picibanil

We devised a new therapeutic modality for multiple hepatocellular carcinoma (HCC) consisting of transarterial immuno-embolization (TIE) using OK-432 and fibrinogen, and applied the treatment to 22 patients with advanced HCC who had been insensitive to TAE, and 21 patients who had no treatment. Nineteen patients had a high AFP level of more than 200 ng/ml. The serum AFP level decreased in 15 patients after TIE, and in 7 patients the AFP level decreased to less than 30%. Furthermore, a marked reduction in tumor size was observed in 69% after TIE. We report 61% 1-year, and 3-year survival rate for cases who had been insensitive to TAE, and 86% 2-year survival rate for those who selected TIE as the first choice. A high fever of more than 38 degrees C occurred in all cases, and hypotension less than 80 mmHg was observed in 42% patients. No deterioration of liver function and no disturbance of the coagulation-fibrinolysis system due to TIE except in one patient with liver failure after TIE.

Topics: Carcinoma, Hepatocellular; Embolization, Therapeutic; Fibrinogen; Humans; Immunotherapy; Infusions, Intra-Arterial; Liver Neoplasms; Picibanil

Immunological and histological analyses were performed on 14 patients with hepatocellular carcinoma treated by transcatheter immunoembolization (TIE) and subsequently by hepatic resection. They were compared with the cases treated by transcatheter arterial embolization (TAE). Exceptionally high plasma levels of inflammatory cytokines, such as IL-6 and IL-8, were noted 3 hours after TIE insults in the majority of the cases. On the contrary, exceptionally high levels of TNF-alpha were also observed in some cases of TIE treatment. In addition, light microscopically, the lytic necrosis of the tumor and massive infiltration of mononuclear cells were the histological characteristics of this treatment. Interestingly, the population of the infiltrates has altered after TIE treatment. It thus consisted mainly of neutrophils in early phase, subsequently of the mixture of lymphocytes, eosinophils, and plasma cells, and finally of lymphocytes. These results may suggest that certain inflammatory responses caused by TIE may play important roles in this new therapeutic modality.

Topics: Carcinoma, Hepatocellular; Chemoembolization, Therapeutic; Fibrinogen; Hepatic Artery; Humans; Interleukin-6; Interleukin-8; Liver Neoplasms; Lymphocyte Subsets; Picibanil; Tumor Necrosis Factor-alpha

We examined whether anti-cancer cells are induced in vivo in hepatocellular carcinoma (HCC) by OK-432. Ten patients with HCC were randomly divided into two groups. The group I patient (n = 5) served as the control. In group II (n = 5), OK-432 was preoperatively administered via the hepatic artery. Tumor infiltrating lymphocytes (TILs) were collected from resected tumors. Cytotoxicity of TILs against K562 cells and Raji cells was studied with phenotypic analysis by flow cytometry. Freshly isolated TILs, whether or not treated with OK-432, showed low cytotoxicity. When TILs were co-cultured with recombinant interleukin-2 (rIL-2), the cytotoxicity was significantly activated in the OK-432 treated group, whereas untreated TILs showed no activation. The natural killer (NK) activity and the lymphokine-activated killer (LAK) activity were depressed in group I after hepatic resection, but patients in group II had no depression. Our data indicate that LAK precursor cells are induced in TILs and the prevention of post-operative immune suppression is made possible by OK-432.

Topics: Carcinoma, Hepatocellular; Cytotoxicity, Immunologic; Hepatic Artery; Humans; Infusions, Intra-Arterial; Iodized Oil; Killer Cells, Lymphokine-Activated; Liver Neoplasms; Lymphocyte Subsets; Lymphocytes, Tumor-Infiltrating; Male; Middle Aged; Picibanil

We investigated the question whether lymphokine-activated killer precursor (pre-LAK) cells are induced by OK-432 in vivo, in hepatocellular carcinoma (HCC). Ten patients with HCC were randomly divided into two groups. In group A (n = 5), OK-432 was pre-operatively administered via the hepatic artery. The group B patients (n = 5) served as controls. Tumor-infiltrating lymphocytes (TILs) were collected from the resected tumors. The cytotoxicity of TILs against natural killer (NK)-sensitive K562 cells and NK-insensitive Raji cells was examined by phenotypic analysis with flow cytometry. Freshly isolated TILs, whether treated with OK-432 or not, showed low cytotoxicity against both tumor cells. However, OK-432 pretreatment increased the T-lymphocyte population of TILs, particularly with interleukin-2 (IL-2) receptor positive cells. When TILs were co-cultured with recombinant interleukin-2 (rIL-2), the cytotoxicity was significantly activated in the OK-432 treated group, while untreated TILs showed no activation (P < 0.05). We postulate that pre-LAK cells are induced by OK-432 in TILs, mainly from the T-lymphocyte population. The possibility that LAK cells can be endogenously induced in HCC if OK-432 and rIL-2 are concomitantly administered needs to be considered for immunotherapy to HCC.

Topics: Adult; Aged; Carcinoma, Hepatocellular; Humans; Immunotherapy; Interleukin-2; Killer Cells, Lymphokine-Activated; Liver Neoplasms; Lymphocytes, Tumor-Infiltrating; Male; Middle Aged; Picibanil; Recombinant Proteins; Stem Cells

An important objective for patients with unresectable hepatocellular carcinoma (HCC) is the development of effective chemotherapy. We administered a combination of biological response modifiers and anticancer agents to 24 patients with unresectable HCC. Each case had an implanted infuser port which was connected to a catheter placed in the hepatic artery for the intraarterial (i.a.) administration of chemotherapy. The following agents were administered to each patient: recombinant interleukin-2 (800,000 JRU/day infused i.a. continuously for 6 days/week); OK-432 (5 KE injected i.a. twice in 4 weeks and i.m. three times per week); Adriamycin (10 mg injected i.a. twice in 4 weeks); cyclophosphamide (300 mg injected i.a. twice in 4 weeks), and famotidine (40 mg/day administered orally). Objective response was assessed according to tumor size measured by computed tomography and angiography before and after treatment. We observed a complete response (CR) in 4, partial response (PR) in 3, minor response (MR) in 7, no change (NC) in 7, and progressive disease (PD) in 3. The response rate (CR+PR+MR) was 58.3%. The overall 2-year survival rate was 52%. The 2-year survival rate of the responders (CR+PR+MR) was 80%, while that of the non-responders (NC+PD) was 0%. There was a significant difference between the responders and non-responders in respect to survival rate (P < 0.05). The percentages of CD25+ cells, CD56+ cells, and Leu7-CD16+ cells and NK activity in the peripheral blood showed a significant increase following the regimen. Serum levels of tumor necrosis factor alpha TNF alpha rose after the initiation of OK-432. TNF alpha levels were higher in the responders than in the non-responders. Adverse effects included high fever (all patients) and severe transient hypotension (15 patients) that was controlled by conservative therapy. Combined immunochemotherapy administered intraarterially may be a new strategy for treating unresectable HCC.

Topics: Aged; Carcinoma, Hepatocellular; Combined Modality Therapy; Cyclophosphamide; Doxorubicin; Famotidine; Female; Humans; Immunity, Cellular; Immunotherapy; Interleukin-2; Killer Cells, Natural; Liver Neoplasms; Lymphocyte Subsets; Male; Middle Aged; Picibanil; Survival Analysis; Tumor Necrosis Factor-alpha

We conducted a randomized, controlled trial comparing 5-fluorouracil (5-FU) with or without biological response modifiers (BRMs) as a maintenance therapy for hepatocellular carcinoma (HCC) after treatment with percutaneous ethanol injection (PEI), transcatheter arterial embolization (TAE) or arterial infusion of antitumor agents (AI). A total of 58 cases of HCC were classified into 4 groups as follows: group I, PSK with 5-FU (n = 15); group II, lentinan with 5-FU (n = 15); group III, OK-432 with 5-FU (n = 12); and group IV, 5-FU alone as the control (n = 16). The mean survival time, mortality rate, time to progression, and T4/T8 ratio of lymphocytes in the peripheral blood were compared among the four groups. There was no significant difference in the background factors among the groups. In group I, the T4/T8 ratio of lymphocytes was reduced after the therapy. No significant difference was found among the groups in terms of the mean survival time, mortality rate, or time to progression. PEI for initial therapy was superior to the other therapies in terms of the mean survival time and mortality rate. These results suggest that the addition of BRM to maintenance therapy with 5-FU exerts no prognostic benefit on HCC patients treated with PEI, TAE, or AI.

Topics: Aged; Carcinoma, Hepatocellular; Chemoembolization, Therapeutic; Combined Modality Therapy; Embolization, Therapeutic; Ethanol; Female; Fluorouracil; Humans; Immunologic Factors; Lentinan; Liver Neoplasms; Male; Middle Aged; Picibanil; Survival Rate

Topics: Administration, Oral; Biological Products; Carcinoma, Hepatocellular; Clinical Trials as Topic; Humans; Infusions, Intra-Arterial; Liver Neoplasms; Mitomycin; Mitomycins; Picibanil; Random Allocation; Tegafur

Radiofrequency ablation (RFA) often results in incomplete ablation for medium-to-large and irregular tumors. To solve this clinical problem, we proposed a new treatment strategy of OK-432 in combination with an anti-programmed cell death protein 1 (αPD-1) antibody for residual tumors after incomplete RFA (iRFA) of hepatocellular carcinoma (HCC).. The effect of OK-432 on immature dendritic cells (iDCs) was evaluated in vitro. A CCK-8 kit and ELISPOT were used to assess the killing effect of OK-432-induced CD8. The combined therapy of OK-432 with αPD-1 antibody induced a strong antitumor immune response, which significantly inhibited the residual tumors after iRFA of HCC. This concept may provide a new treatment strategy to increase the curative efficacy of RFA for medium-to-large and irregular HCCs.. The data of this study are available from the corresponding author on reasonable request.

Topics: Animals; Antibodies; Carcinoma, Hepatocellular; CD8-Positive T-Lymphocytes; Liver Neoplasms; Mice; Mice, Inbred Strains; Neoplasm, Residual; Picibanil; Radiofrequency Ablation

To investigate the efficacy of an injectable hydrogel loaded with lysed OK-432 (lyOK-432) and doxorubicin (DOX) for residual liver cancer after incomplete radiofrequency ablation (iRFA) of hepatocellular carcinoma (HCC), and explore the underlying mechanism.. The effect of OK-432 and lyOK-432 was compared in activating dendritic cells (DCs). RADA16-I (R) peptide was dissolved in a mixture of lyOK-432 (O) and DOX (D) to develop an ROD hydrogel. The characteristics of ROD hydrogel were evaluated. Tumor response and mice survival were measured after different treatments. The number of immune cells and cytokine levels were measured, and the activation of cGAS/STING/IFN-I signaling pathway in DC was evaluated both in vitro and in vivo.. LyOK-432 was more effective than OK-432 in promoting DC maturation and activating the IFN-I pathway. ROD was an injectable hydrogel for effectively loading lyOK-432 and DOX, and presented the controlled-release property. ROD treatment achieved the highest tumor necrosis rate (p < 0.001) and the longest survival time (p < 0.001) compared with the other therapies. The ROD group also displayed the highest percentages of DCs, CD4. The novel ROD peptide hydrogel induced an antitumor immunity by activating the STING pathway, which was effective for treating residual liver cancer after iRFA of HCC.

Topics: Animals; Carcinoma, Hepatocellular; CD8-Positive T-Lymphocytes; Cytokines; Doxorubicin; Hydrogels; Liver Neoplasms; Mice; Picibanil; Radiofrequency Ablation

Preoperative transarterial immunoembolization (TIE) for hepatocellular carcinoma (HCC) is effective for preventing recurrence. We aimed to investigate the intratumoral and peritumoral M1 macrophage-induced immune response following TIE treatment.. We compared 13 patients treated with TIE between 2003 and 2009 (TIE group) and 13 patients treated with surgery alone during the same period of time at our institute (control group) using an immunohistological study with CD68 and CD163 antibodies.. No significant differences in clinicopathological characteristics, except for surgical time, were observed between the two groups. The 3-year recurrence-free survival outcome of the TIE group was quite different from that of the control group (100% vs. 38.5%, P = 0.034). In the histological investigation, lytic necrosis and coagulation necrosis of the main tumor along with the presence of multinuclear giant cells were observed in 10 of the 13 patients in the TIE group. The immunohistological study showed that not only the numbers of intratumoral CD68(+) cells, but also the numbers of intratumoral and peritumoral CD8(+) cells were significantly increased in the TIE group.. The suppression of tumor recurrence induced by preoperative TIE might be induced by intratumoral M1 macrophages that are activated by OK-432 and fibrinogen.

Topics: Aged; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Antineoplastic Agents; Carcinoma, Hepatocellular; Catheterization, Peripheral; Chemoembolization, Therapeutic; Disease-Free Survival; Female; Follow-Up Studies; Humans; Injections, Intra-Arterial; Liver; Liver Neoplasms; Macrophages; Male; Middle Aged; Picibanil; Receptors, Cell Surface; Retrospective Studies; Treatment Outcome

Despite curative locoregional treatments for hepatocellular carcinoma (HCC), tumour recurrence rates remain high. The current study was designed to assess the safety and bioactivity of infusion of dendritic cells (DCs) stimulated with OK432, a streptococcus-derived anti-cancer immunotherapeutic agent, into tumour tissues following transcatheter hepatic arterial embolization (TAE) treatment in patients with HCC. DCs were derived from peripheral blood monocytes of patients with hepatitis C virus-related cirrhosis and HCC in the presence of interleukin (IL)-4 and granulocyte-macrophage colony-stimulating factor and stimulated with 0·1 KE/ml OK432 for 2 days. Thirteen patients were administered with 5 × 10⁶ of DCs through arterial catheter during the procedures of TAE treatment on day 7. The immunomodulatory effects and clinical responses were evaluated in comparison with a group of 22 historical controls treated with TAE but without DC transfer. OK432 stimulation of immature DCs promoted their maturation towards cells with activated phenotypes, high expression of a homing receptor, fairly well-preserved phagocytic capacity, greatly enhanced cytokine production and effective tumoricidal activity. Administration of OK432-stimulated DCs to patients was found to be feasible and safe. Kaplan-Meier analysis revealed prolonged recurrence-free survival of patients treated in this manner compared with the historical controls (P = 0·046, log-rank test). The bioactivity of the transferred DCs was reflected in higher serum concentrations of the cytokines IL-9, IL-15 and tumour necrosis factor-α and the chemokines CCL4 and CCL11. Collectively, this study suggests that a DC-based, active immunotherapeutic strategy in combination with locoregional treatments exerts beneficial anti-tumour effects against liver cancer.

Topics: Aged; Aged, 80 and over; Antineoplastic Agents; Carcinoma, Hepatocellular; Combined Modality Therapy; Cytokines; Dendritic Cells; Disease-Free Survival; Embolization, Therapeutic; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Hepatitis C; Humans; Immunotherapy, Active; Interleukin-4; Liver Neoplasms; Male; Middle Aged; Monocytes; Neoplasm Recurrence, Local; Picibanil; Radiography

Therapeutic vaccination with dendritic cells (DCs) pulsed with tumor cell lysate vaccine (H-D) represents an attractive approach for hepatocellular carcinoma (HCC) treatment. However, the efficacy of this approach is not most satisfactory for the low levels of T helper 1 (Th1)-type cytokines secretion and weak T cell responses. In this study, in order to increase the potency of H-D, two tandem repeats of microbial HSP70 peptide epitope 407-426 (2mHSP70(407-426), M2) which has been demonstrated to be effective in enhancing DC maturation were applied. The DC vaccine (HM-D) which was HCC tumor cell lysate pulsed with M2 was developed. Nevertheless, the immunotherapeutic effect was still not satisfactory enough even some promotion was obtained. Therefore, OK-432 (OK), which is a useful anti-cancer agent and effectively in stimulating DC maturation, was introduced to HM-D. Our results demonstrated that treatment with the improved DC vaccine which was tumor cell lysate pulsed with M2 and OK (HMO-D), compared with H-D and HM-D, significantly increased cell surface markers (MHC-I and II, CD40, CD80, CD86 and CD11c) expression on DCs, enhanced Th1-type cytokines (IL-12, TNF-α and IFN-γ) production but not Th2-type cytokine (IL-5) production, induced remarkable high levels of lymphocytes proliferation and CD8(+) cytotoxic T-lymphocyte (CTL). Furthermore, immunization with HMO-D effectively reduced tumor progression and enhanced the survival of mice with H22 tumors. Besides, we also found that the capability of M2 in inducing the Th1 cytokines was stronger than OK. In view of these results, HMO-D vaccination provided a novel immunotherapeutic approach for the treatment of HCC.

Topics: Animals; Cancer Vaccines; Carcinoma, Hepatocellular; Cytokines; Dendritic Cells; Epitopes; HSP70 Heat-Shock Proteins; Liver Neoplasms, Experimental; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Picibanil; T-Lymphocytes, Cytotoxic; Tandem Repeat Sequences; Treatment Outcome

Suppressor of cytokine signaling 1 (SOCS1) plays a critical role in antitumor immunity. Down-regulating SOCS1 in antigen-presenting dendritic cells (DCs) could enhance antigen-specific antitumor immunity. This study was to investigate the antigen-specific antitumor effect and mechanism of DCs with siRNA-mediated inhibition of SOCS1, stimulated by OK-432 and pulsed with hepatocellular carcinoma cell line HepG2 antigens.. The expression of SOCS1 in immature DCs was down-regulated by RNA interference (RNAi). DCs were pulsed with lysate of HepG2 cells and stimulated with OK-432. The morphology of DCs was observed under converted phase microscopy. Phenotypic changes in cells after stimulation were characterized by flow cytometry (FCM). The Alamar Blue assay was adopted to evaluate the activation and proliferation of autologous lymphocytes induced by mature DCs. The cytotoxicity of cytotoxic T lymphocytes (CTLs) elicited by modified DCs to HepG2, EC109 and K562 cells was tested by the lactate dehydrogenase (LDH) assay.. Cells displaying a typical morphology and phenotypic properties of mature DCs were obtained successfully. The expression of SOCS1 in DCs was down-regulated by SOCS1 RNAi. Mature DCs showed high expressions of CD80, CD83, CD86, and HLA-DR. Pulsing of DCs with lysate of HepG2 had no influence on the phenotypic properties of DCs. Down-regulating SOCS1 expression enhanced the maturation of DCs. The modified DC tumor vaccine stimulated the proliferation of autologous lymphocytes effectively, and the proliferation rate of T cells was (110.7+/-22.2)%. After being activated by modified DCs, TCLs exerted a specific and effective killing effect on HepG2 cells, but not on EC109 and K562 cells.. Mature DCs could induce antigen-specific antitumor immunity against hepatocellular carcinoma after silencing of SOCS1 by siRNA, stimulation by OK-432 and pulsing of DCs with HepG2 cell antigens.

Topics: Antineoplastic Agents; Cancer Vaccines; Carcinoma, Hepatocellular; Cell Line, Tumor; Cytotoxicity, Immunologic; Dendritic Cells; Humans; Liver Neoplasms; Lymphocyte Activation; Picibanil; RNA, Small Interfering; Suppressor of Cytokine Signaling 1 Protein; Suppressor of Cytokine Signaling Proteins; T-Lymphocytes, Cytotoxic

We report a 51-year-old man with a ruptured hepatocellular carcinoma (HCC). He was admitted to the hospital with abdominal pain and distension. Imaging studies revealed massive ascites, liver cirrhosis, and a 3-cm tumor at the inferior edge of the medial segment of the liver, with adhesions to the greater omentum. Abdominal paracentesis showed bloody ascites, and the patient was diagnosed with a ruptured HCC. OK-432, an immunomodulatory agent prepared from an attenuated strain of Streptococcus pyogenes, was injected (10 KE) into the peritoneal cavity four times within 1 week; the massive ascites disappeared, and the serum alpha-fetoprotein (AFP) level decreased to within the normal limits. Afterwards, he underwent a curative operation for HCC. His postoperative course was uneventful and he was discharged from the hospital on the twenty-second postoperative day. He had shown no evidence of recurrence or metastases at the time he died of hepatic failure related to alcohol abuse 9 months after the operation.

Topics: Antineoplastic Agents; Carcinoma, Hepatocellular; Humans; Injections, Intraperitoneal; Liver Neoplasms; Male; Middle Aged; Picibanil; Rupture, Spontaneous

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.. 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.. 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.. 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.

Topics: Animals; Carcinoma, Hepatocellular; Disease Models, Animal; Interferon-gamma; Killer Cells, Natural; Liver; Liver Neoplasms, Experimental; Lymphocyte Subsets; Male; Picibanil; Rats; Rats, Wistar; Survival Analysis

Recurrence of hepatocellular carcinoma (HCC) is frequent, even after apparently curative resection. Preoperative transcatheter arterial chemoembolization (TAE) does not improve disease-free survival after hepatic resection. We previously reported the potential usefulness of transarterial immunoembolization (TIE), a newly developed arterial embolization technique using OK-432 and fibrinogen, as preoperative treatment. In this study, we further investigated the effect of TIE by histologic examination of the resected specimens and compared it with conventional TAE in a prospective nonrandomized manner. Thirty-nine patients underwent TIE (n = 17) or TAE (n = 22) before curative hepatectomy for HCC. Transarterial immunoembolization was performed according to the standard protocol using OK-432, fibrinogen, and thrombin. Histologic changes in cancerous and noncancerous liver tissues were examined at different stages after TIE. Histologic grading of cancer cell injury according to the modified Shimosato criteria (Grades 0-IV, in increasing order of severity of cell injury) and postoperative disease-free survival were compared between the two groups. Based on the results of histopathology, TAE was more effective than TIE against the main tumor. In contrast, TIE was significantly more effective than TAE against extracapsular invasion and intrahepatic metastasis. Disease-free survival after hepatectomy tended to be better in patients pretreated with TIE than TAE. Postoperative tumor recurrences in the TIE group (n = 4) occurred in the nontreatment regions, whereas tumor recurrences in TAE group developed mostly (8 of 11 patients) in treated liver regions. Based on results of histologic examination, TIE seems to be more effective than conventional TAE against extracapsular invasion and intrahepatic metastasis. Data for disease-free survival and recurrence site suggest TIE may be a useful preoperative treatment.

Topics: Antineoplastic Agents; Carcinoma, Hepatocellular; Catheterization; Disease-Free Survival; Embolization, Therapeutic; Female; Fibrinogen; Hepatectomy; Hepatic Artery; Humans; Liver; Liver Neoplasms; Male; Middle Aged; Neoadjuvant Therapy; Neoplasm Recurrence, Local; Picibanil; Preoperative Care; Prognosis; Thrombin; Time Factors

Surgical resection, transcatheter arterial embolization (TAE) and percutaneous ethanol injection therapy (PEIT) are effective for hepatocellular carcinoma (HCC), but the recurrence rate is high. We have devised a new therapy of transarterial immuno-embolization (TIE) with OK-432, fibrinogen and thrombin, and 2 cases are reported. Case 1: A 78-year-old Japanese male with HCC (diameter, 4 cm in subsegment 5) received TIE. The tumor size was markedly decreased, and the patient survived for more than 3 years without recurrence. Case 2: A 61-year-old Japanese male with HCC (diameter, 4.5 cm in segment 5) received hepatic subsegmentectomy following TIE. Histological examination of resected specimens following TIE showed massive infiltration of mononuclear cells in the main tumor. Tumor recurrence had developed three times thereafter, but was effectively treated by TIE. TIE may be an effective therapy for HCC.

Topics: Aged; Carcinoma, Hepatocellular; Embolization, Therapeutic; Fibrinogen; Hepatic Artery; Humans; Immunotherapy; Iodized Oil; Liver Neoplasms; Male; Middle Aged; Picibanil; Thrombin

The antitumor effects of immunotherapy using streptococcal preparations (OK-432), recombinant granulocyte-colony-stimulating factor (rG-CSF) and recombinant interleukin-2 (rIL-2) were examined for human hepatocellular carcinoma (HCC). Following subcutaneous injection of OK-432 (2 KE) and rG-CSF (50-60 microg), low-dose intratumoral administration of OK-432 (3-12 KE) was performed. Thereafter, 2 x 10(5) JRU of rIL-2 was subcutaneously injected. This therapeutic regimen was repeated twice. Serum alpha-fetoprotein levels were markedly decreased in three of seven patients with HCC by this treatment. Post-therapeutic histological examination revealed that trabecular cords or pseudoglandular arrangements of tumor cells were completely disordered in all cases and that extensive infiltration of lymphocytes into the tumor stroma was present in five cases. The number of CD4- and CD57-positive cells among tumor-infiltrating lymphocytes after immunotherapy was significantly higher than that in patients without immunotherapy (P <0.01). These findings suggest that even a small intratumoral injection of OK-432 can induce extensive infiltration of helper/inducer and natural killer cells into the tumor stroma when combined with subcutaneous injection of OK-432, rG-CSF and rIL-2 and that these cells might play important roles in tumor cytotoxicity.

Topics: Aged; alpha-Fetoproteins; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Hepatocellular; Drug Administration Schedule; Female; Granulocyte Colony-Stimulating Factor; Humans; Immunohistochemistry; Immunotherapy; Interleukin-2; Leukocyte Count; Liver Neoplasms; Male; Middle Aged; Neutrophils; Picibanil; Recombinant Proteins

The antitumor activity of combination therapy with traditional Chinese medicines and OK432 (Streptococcus pyogenes) for cancer patients was investigated. Excellent antitumor activity of this treatment was achieved in one patient with hepatocellular carcinoma. The present report describes the clinical course of this patient and examines the contribution of production of tumor necrosis factor (TNF) and interferon-gamma (IFN). Endogenous production of TNF could be observed after drip intravenous injection of OK 432 in the serum of patients treated by previous oral administration of traditional Chinese medicines. The serum levels of IFN were very low and remained at almost undetectable levels under these conditions. The selective use of immunostimulants such as traditional Chinese medicines may be of value in combination with other therapies such as drip infusion of OK 432, in the treatment of advanced cancer or of aged patients because of the low toxicity. One patient out of 12 revealed a partial response as assessed by the antitumor activity. However, with this treatment, patients did become free from pain and a good performance status was supported.

Topics: Aged; Aged, 80 and over; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Hepatocellular; Drugs, Chinese Herbal; Female; Humans; Interferon-gamma; Interleukin-1; Liver Neoplasms; Lymphocyte Count; Male; Picibanil; T-Lymphocyte Subsets; Tumor Necrosis Factor-alpha

The prognosis of patients with multiple hepatocellular carcinoma (HCC) remains disappointing. In this study, we devised a new therapeutic modality for HCC consisting of transarterial immunoembolization (TIE) using OK-432 and fibrinogen and then analyzed the preliminary results. In the first series, we applied the treatment to 19 patients with advanced HCC who had proved to be insensitive to several previous conventional treatments. In all, 14 patients (74%) with unresected HCC have currently survived for between 2 and 16 months after TIE. The remaining 5 patients died at 17, 14, 8, 7, and 4 months after TIE. The serum levels of tumor markers decreased in all of the patients, and a marked reduction in tumor size was observed in six patients after TIE. A high fever occurred in all cases, and abdominal pain and loss of appetite were also observed after TIE. However, deterioration of liver function was negligible. After confirmation of the safety of this method, we started a second study series in which this TIE treatment was selected as the first choice. Six patients have been treated to date. All patients in this group underwent hepatic resection at 6-48 days following TIE. Histological examination of the resected specimens following TIE showed massive infiltration of mononuclear cells around tumor cell nests and lytic necrosis as well as coagulation necrosis of the main tumor and the intrahepatic metastases. In conclusion, our results indicate that TIE may be a safe and promising therapy for patients with HCC.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Hepatocellular; Embolization, Therapeutic; Female; Fibrinogen; Humans; Immunotherapy; Injections, Intra-Arterial; Liver Neoplasms; Male; Middle Aged; Picibanil; Prognosis

We report here a case of hepatocellular carcinoma (HCC) with multiple lung metastases, which were disappeared by treatment of OK-432. The patient was a 65-year-old man and was diagnosed in 1986 with a small (17 x 11 mm) HCC in the anterior-superior segment of the liver. A part of the right hepatic lobe including the tumor was surgically removed, and transarterial injections of adriamycin (10 mg/week) and subcutaneous injections of OK-432 (10 KE/week) were given. Two and a half years later, recurrence of HCC in the liver and its invasion to vena cava inferior (IVC) were found. OK-432 administration was then stopped and percutaneous ethanol injection therapy (PEIT) was performed 10 times. Six months later, the PEIT was effective and the liver tumor with IVC invasion diminished. However, multiple lung metastases were visible on roentgenograms of the chest, and serum alphafetoprotein (AFP) concentration increased to 50,000 ng/ml. The OK-432 treatment resumed. After 6 months of OK-432 treatment, the multiple lung metastases were disappeared and the serum AFP level decreased to 100 ng/ml. At present, the patient is surviving without any sign of recurrence in either the liver or the lung. The clinical course of this case suggests that OK-432 might have effectively treated lung metastases of HCC, although the exact mechanisms are at present unclear.

Topics: Aged; alpha-Fetoproteins; Carcinoma, Hepatocellular; Humans; Liver Neoplasms; Lung Neoplasms; Male; Picibanil

The efficacy of transarterial immuno-embolization therapy (TIE) was examined in six operable patients with hepatocellular carcinoma (HCC). We administered OK-432, fibrinogen (30 mg/ml) and thrombin (1 U/ml) through a catheter which was inserted into the tumor-feeding artery. In all patients with a high level of tumor markers (AFP and PIVKA-II), the level decreased promptly to less than the pretreatment level after TIE therapy. The therapy has not caused any serious side effects. No disturbance of the coagulation-fibrinolysis system due to TIE was observed in any patient. Histological examination of resected specimens following TIE showed massive infiltration of mononuclear cells around tumor cell nests, and lytic necrosis as well as coagulation necrosis of the main tumor and the intrahepatic metastases. Our results indicate that TIE may be an effective and promising modality for HCC patients.

Topics: Aged; Carcinoma, Hepatocellular; Chemoembolization, Therapeutic; Combined Modality Therapy; Female; Fibrinogen; Humans; Iodized Oil; Liver Neoplasms; Male; Middle Aged; Picibanil; Thrombin

Human neutrophil-mediated oxidative processes against a human hepatoma cell line, HCC-M, was visualized at the cellular level by using a silicon-intensified target camera and subsequently processing with a computer-assisted digital-imaging processor. Neutrophils were activated by a streptococcal preparation, OK-432. A hydroperoxide-sensitive tracer, dichlorofluorescein diacetate, was loaded in HCC-M and temporal and spatial changes of lipid peroxides in this cell after addition of stimulated neutrophils were analyzed. The luminol-dependent chemiluminescence activity of neutrophils was significantly enhanced and continued for at least 2 hr by stimulation with OK-432, and its activity was shown to be accumulated at the site where a neutrophil attached with HCC-M. The intensity of dichlorofluorescein fluorescence in HCC-M rapidly increased after adding stimulated neutrophils, and their reaction was significantly attenuated by superoxide dismutase. The number of non-viable cells was increased as the dichlorofluorescein fluorescence increase. It is suggested that oxidative stress may play an important role in neutrophil-mediated tumor-cell damage.

Topics: Carcinoma, Hepatocellular; Fluoresceins; Humans; Liver Neoplasms; Luminescent Measurements; Neutrophils; Oxidation-Reduction; Picibanil; Tetradecanoylphorbol Acetate; Tumor Cells, Cultured

The efficacy of transarterial immuno-embolization therapy (TIE) was investigated in 7 patients with multiple hepatocellular carcinoma (HCC). We administered OK-432 (10 KE), fibrinogen (30 mg/ml) and thrombin (1 U/ml) as a new treatment for HCC with intrahepatic metastasis. In all patients with a high level of AFP, the AFP level decreased promptly less than the pretreatment level after TIE therapy. High fever in all cases, epigastralgia in 6 cases and appetite loss in 3 cases were observed after TIE therapy. Our results indicate that this therapy may be a safe and effective method in HCC patients with intrahepatic metastasis.

Topics: Administration, Oral; Aged; Carcinoma, Hepatocellular; Embolization, Therapeutic; Fibrinogen; Fluorouracil; Hepatic Artery; Humans; Immunotherapy; Iodized Oil; Liver Neoplasms; Male; Middle Aged; Picibanil; Thrombin

A 66-year-old woman was hospitalized in a state of shock with rupture of hepatocellular carcinoma and multiple pulmonary metastasis. Her bleeding was successfully controlled by emergency transcatheter arterial embolization with Lipiodol (Lp-TAE). Treatments with UFT, OK-432 and two additional Lp-TAE caused the disappearance of pulmonary metastasis with AFP levels decreased and natural killer cell activity increased. The patient died one and a half years after the emergency Lp-TAE. The disappearance of pulmonary metastatic lesions seemed to be caused by improvement of the patient's immunity, which related to the regression of primary tumor after Lp-TAE. It was suggested that Lp-TAE is worth undertaking even in rupture of hepatocellular carcinoma with remote metastatic lesions.

Topics: Aged; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Hepatocellular; Chemoembolization, Therapeutic; Doxorubicin; Female; Hepatic Artery; Humans; Iodized Oil; Liver Neoplasms; Lung Neoplasms; Mitomycin; Picibanil; Prognosis; Remission Induction; Rupture, Spontaneous; Tegafur; Uracil

To increase antitumor effects of transcatheter arterial embolization therapy (TAE) for hepatocellular carcinoma (HCC), immunochemo-embolization therapy via hepatic artery was performed with a mixture of doxorubicin and iodized oil (LPD) following a mixture of gamma-IFN, OK-432 and gelatin sponge, and then a mixture of actinomycin D and gelatin sponge. Three patients with HCC were treated by this procedure. One patient had tumor thrombus in inferior vena cava (IVC). The serum alpha fetoprotein (AFP) levels before this procedure ranged from 66 ng/ml to 8,360 ng/ml. Following this procedure, the serum AFP levels began to decrease for 1-3 months, then increased for 3-6 months, and again suddenly decreased under 10 ng/ml in two cases after initial procedure. The serum AFP levels of two cases revealed under 10 ng/ml for 9-18 months. CT after 2 weeks to 3 months of this procedure showed a low-density area around LPD-uptaking tumor and after 1-8 months decreased tumor in size with diminishing of the low-density area. Therapy for the main tumor of one case with tumor thrombus of IVC proved to be effective, but it was not effective for tumor thrombus of IVC. The reasons that the serum AFP level increased after decreasing for 1-3 months and then fell below 10 ng/ml following this procedure, may be some kinds of immunological antitumor effects produced by endogenous cytokines.

Topics: Aged; alpha-Fetoproteins; Carcinoma, Hepatocellular; Dactinomycin; Doxorubicin; Female; Humans; Interferon-gamma; Iodized Oil; Liver Neoplasms; Male; Middle Aged; Picibanil; Tomography, X-Ray Computed

The endogenous induction of tumor necrosis factor serum (TNS) for cancer immunotherapy was undertaken in the immediate postoperative period using Lentinan as the primer and OK-432 as the inducer. The changes in several immunological markers of the blood were assayed and compared with a control group to clarify the effects of this treatment. Plasma TNF-alpha levels were elevated two to three hours after eliciting treatment. The neutrophil count was elevated on the 7th postoperative day (POD) and the natural killer (NK) cell activity was transiently suppressed on the 1st POD, but NK cells possessing a high activity (Leu7-CD16+) were preserved until the 7th POD. Helper/inducer (CD4+) and killer cells (CD8+ CD11-) tended to increase, and suppressor (CD8 bright+ CD11+) cells tended to decrease in the induction group. There was no difference in the levels of prostaglandin E2 (PGE2) between the groups, but a marked elevation of interferon-gamma was evident on the 1st POD in the induction group. This treatment may be useful as postoperative adjuvant immunotherapy for cancer due to its ability to induce cytokines and activate host immune mechanisms.

Topics: Aged; Antigens, CD; Biomarkers, Tumor; Carcinoma, Hepatocellular; CD4-Positive T-Lymphocytes; Esophageal Neoplasms; Female; Humans; Immunotherapy; Interferons; Lentinan; Leukocyte Count; Liver Neoplasms; Male; Middle Aged; Pancreatic Neoplasms; Picibanil; Postoperative Period; Remission Induction; Stomach Neoplasms; T-Lymphocytes, Regulatory; Tumor Necrosis Factor-alpha

OK-432 and etoposide were administered intravenously to a patient with liver metastasis of ovarian cancer. OK-432 induced IFN-gamma and TNF. SuPS and PPD skin reaction became positive after the therapy. The size of metastatic lesions was reduced significantly (partial response). It is suggested that the combination therapy of OK-432 and etoposide may be effective to the liver metastasis of ovarian cancer.

Topics: Adult; Carcinoma, Hepatocellular; Etoposide; Female; Humans; Injections, Intravenous; Liver Neoplasms; Ovarian Neoplasms; Picibanil

Unresectable hepatocellular carcinoma (HCC) has a poor prognosis and little sensitivity to anticancer agents. We planned a combination immunotherapy, associating with rIL-2 continuous injection. Combination immunotherapy consists of 5 different biological response modifiers (BRM), i.e., rIL-2, OK-432, adriamycin (ADR), cyclophosphamide (Cy), and famotidine (Fa). rIL-2, OK-432 and ADR were administered from implantable infuser port connecting to a catheter which was injected into the hepatic artery via the gastroduodenal artery under laparotomy. OK-432, Cy, and Fa were also administered systemically. From 1988 to 1990, seven patients with unresectable HCC and two patients who underwent curative resection were treated by this therapy. The objective responses were evaluated by CT and angiography. This combination immunotherapy produced 3 cases of complete response and 2 of minor response. Immunological monitoring of lymphocyte subsets and NK activity of peripheral blood mononuclear cells was also performed. NK activity was significantly augmented and the percentage of Leu 7(-) NCD 16(+) cells increased during this therapy. Serious toxicity, but transient high fever and hypotension, was not observed during this therapy. All of these patients left the hospital within two weeks and returned to their previous job or similar life activity. These results suggest that this combination immunotherapy is worth performing in further clinical trials for hepatocellular carcinoma.

Topics: Antineoplastic Combined Chemotherapy Protocols; Biological Products; Carcinoma, Hepatocellular; Cyclophosphamide; Doxorubicin; Famotidine; Female; Hepatic Artery; Humans; Infusions, Intra-Arterial; Injections, Intramuscular; Interleukin-2; Liver Neoplasms; Male; Middle Aged; Picibanil; Quality of Life; Remission Induction

Antitumor effect of TNF has been demonstrated to be increased with some kinds of anticancer agents. We reported antitumor effect of hepatic endogenous TNF induced with gamma-IFN and OK-432 for hepatocellular carcinoma (HCC). To increase antitumor effect of transcatheter arterial embolization (TAE), hepatic arterial chemoembolization was performed with a mixture of gamma-IFN, OK-432 and gelatin sponge following a mixture of Doxorubicin and iodized oil (LPO) on the first time. Serum alpha-fetoprotein decreased from 18,903 ng/ml to 470 ng/ml but elevated three months after these procedures. Following the above procedure, hepatic arterial embolization with a mixture of gelatin sponge and Actinomycin D as an inhibitor of RNA was given the second time. Serum alpha-fetoprotein decreased under 5 ng/ml and computed tomography revealed decreased tumor size and low density area following this second procedure. Hepatic arterial chemoembolization with a mixture of hepatic induction of endogenous TNF and anticancer agents may well be beneficial for survival of patient with HCC.

Topics: Aged; Antineoplastic Combined Chemotherapy Protocols; Biological Products; Carcinoma, Hepatocellular; Dactinomycin; Delayed-Action Preparations; Doxorubicin; Drug Administration Schedule; Embolization, Therapeutic; Gelatin Sponge, Absorbable; Hepatic Artery; Humans; Infusions, Intra-Arterial; Interferon-gamma; Iodized Oil; Liver Neoplasms; Male; Picibanil; Tumor Necrosis Factor-alpha

Intratumor injection of OK-432, a biological response modifier, in the treatment of small HCC was studied in 7 inoperable patients. After evaluation with ultrasound (US), computed tomography (CT), angiography and US-guided biopsy, implantation of a steel coil in the tumor, intratumor injection was performed under US guidance. After completion of the treatment, liver biopsy and image studies were again done to evaluate the extent of tumor necrosis. One patient was alive and well without recurrence 19 months after treatment. Four had recurrent tumors at different site of the liver 4 months, 9 months, 9 months and 8 months later. Two died of progressive malignancy 3 months and 8 months later. In the 6 patients with elevated serum alpha-fetoprotein (AFP) levels, 4 had decreased AFP after treatment, and the 2 mortalities had steadily increased AFP. The most common side effects are fever and chills. Transient abdominal pain with elevated transaminase activities, cough with hemoptysis, and vomiting were seen in 1 case each. After treatment, the biopsy specimens showed total necrosis of HCC. Although the T4/T8 ratio of peripheral blood was increased as compared with that before treatment in 4 cases, peritumoral cytotoxic T lymphocyte and monocyte infiltration were seen in one specimen only, and another 7 examined specimens showed negative staining with monoclonal antibodies of T cells. We conclude that intratumor injection of OK-432 is an alternative treatment for small HCC in inoperable cases. The effectiveness may be due to the direct tumoricidal mechanism of OK-432.

Topics: Aged; Carcinoma, Hepatocellular; Female; Humans; Injections, Intralesional; Leukocytes, Mononuclear; Liver Neoplasms; Lymphocyte Subsets; Male; Middle Aged; Picibanil; T-Lymphocytes, Cytotoxic

Lymphokine-activated killer (LAK) activity of peripheral blood mononuclear cells (PBMC) from 33 patients with hepatocellular carcinoma was significantly decreased compared with that of healthy volunteers. There was less LAK activity in PBMC from patients with larger tumours (5 cm or more in diameter) than in patients with smaller tumours (under 5 cm in diameter). In 8 out of 20 patients with larger tumours there was none or little LAK activity. Flow cytometry revealed that the percentage of Leu11b+ cells in PBMC was lower in patients than in normal volunteers, and was lowest in patients with larger tumours. 10 patients with hepatocellular carcinoma were treated with intratumoral injection of OK432. LAK activity was enhanced after treatment in 7 cases, and the percentage of Leu11b+ cells was increased. Enhancement of LAK activity in response to OK432 was more significant in patients with smaller rather than larger tumours. Of the 7 high LAK responders, 4 showed 50-100% tumour regression at 6-9 weeks after injection.

Topics: Carcinoma, Hepatocellular; Cytotoxicity, Immunologic; Female; Flow Cytometry; Humans; Killer Cells, Lymphokine-Activated; Leukocyte Count; Liver Neoplasms; Male; Middle Aged; Picibanil

Antitumor effect and reduction of tumor size by some cytokines as Biological Response Modifier have been demonstrated by various studies. Endogenous tumor necrosis factor is produced from macrophage. To increase the antitumor effect of transcatheter arterial embolization (TAE) in hepatocellular carcinoma (HCC), we treated 7 HCC patients with endogenous tumor necrosis factor (ETNF) which was induced by hepatic arterial injection of gamma-IFN (1.0-3.0 X 10(6) IU) as priming agent and OK-432 (2-5 KE) as triggering agent. TAE was performed with Lipiodol, ADM and gelatin sponge on 3-10 days after the induction of ETNF. TNF activity was detected in 2 cases and suspected to depend on the dose of gamma-IFN and OK-432. Serum alpha-Fetoprotein levels after the injection of ETNF began to decrease from 3-30 days in 5 patients and remained unchanged in 2 cases. Serum alpha-Fetoprotein levels after TAE with the induction of ETNF were decreased 1-5 months in 5 cases. Reduced size and low-density area on CT scan in 3 advanced cases after these procedures were no different from those of HCC patients treated with TAE alone. In one of two inoperable cases with a single mass lesion in the liver, CT scan after one more added TAE following these procedures showed a low-density area around the Lipiodol uptaking tumor, indicating obstruction of the peripheral portal vein. CT scan of another case revealed low density around Lipiodol in the tumor, which showed complete necrotic change. In all cases, middle-grade fever and hypotension were seen transiently, but these subsided by symptomatic treatment. The antitumor effect of TAE in HCC might be enhanced with ETNF induced by hepatic arterial injection of a low dose of gamma-INF and OK-432.

Topics: Aged; alpha-Fetoproteins; Carcinoma, Hepatocellular; Combined Modality Therapy; Doxorubicin; Embolization, Therapeutic; Female; Hepatic Artery; Humans; Infusions, Intra-Arterial; Interferon-gamma; Liver Neoplasms; Male; Middle Aged; Picibanil; Tumor Necrosis Factor-alpha

The relationship between nonspecific cytotoxic activity of spleen cells and the resistance against the graft challenge of a human hepatoma cell line (HCC-M) was investigated in nude mice. Two administrations of an immunopotentiator, OK-432 or human interleukin-2, prior to the subcutaneous inoculation of HCC-M cells, which was performed 24 h after the last administration, significantly inhibited the tumor development in terms of rate of tumor take and tumor size. This effect was abrogated by simultaneous administration of an anti-asialo GM1 (ASGM1) antiserum. There was a significant inverse correlation between tumor volume and spleen cell cytotoxicity which was determined at the time of HCC-M cell inoculation against a YAC-1 or HCC-M target. Spleen cell cytotoxicity enhanced by these immunopotentiators could not completely be abolished by in vitro treatment with ASGM1 and complement. This result suggests that effector cells of the enhanced cytotoxicity consist of heterogeneous cells including both ASGM1+ natural killer cells and other nonselective cytotoxic cells. These results suggest that nonspecific cytotoxic cells play crucial roles in the resistance against tumor cell challenge and that the total level of cytotoxic activity of these cells at the time of tumor cell challenge is a key factor which determines tumor development.

Topics: Animals; Biological Products; Carcinoma, Hepatocellular; Cell Line; Cytotoxicity, Immunologic; Female; Graft Rejection; Humans; Interleukin-2; Liver Neoplasms; Male; Mice; Mice, Nude; Neoplasm Transplantation; Picibanil; Spleen

In order to evaluate the combination therapy for liver and bile tract cancer, the effects of anticancer drugs and hyperthermia were observed using cultured human cancer cell lines. In the case of gall bladder cancer cell line (NOZ), combination of adriamycin and hyperthermia showed more effective inhibition for cell proliferation than MMC + hyperthermia and 5-FU + hyperthermia. Hepatocellular carcinoma cell line (JHH-4) showed remarkable inhibition of cell growth and secretion of albumin by combination treatment of adriamycin and hyperthermia. Morphologically, JHH-4 cells were enlarged and the nucleus was also enlarged with combination adriamycin and hyperthermia by phase contrast microscopy. Cytoskeleton of JHH-4 cells became irregular and intercellular borderline was unclear by plasma polymerization replica method (PPRM). The effects of BRM (OK-432 and TNF) on HCC cell lines was also investigated. OK-432 directly inhibited proliferation of JHH-4 cells. We observed internalization of OK-432 by JHH-4 cells with TEM and 16-mm movie. TNF showed various effects on human HCC cell lines. Proliferation of two cell lines was inhibited, and one tended to be enhanced after the addition of TNF to the medium. Hyperthermia influenced the effects of TNF to HCC cell lines. We think that this paper is a very significant study for improving the therapy for hepato-biliary cancers.

Topics: Antineoplastic Agents; Carcinoma, Hepatocellular; Cell Line; Combined Modality Therapy; Doxorubicin; Fluorouracil; Gallbladder Neoplasms; Humans; Hyperthermia, Induced; In Vitro Techniques; Liver Neoplasms; Mitomycin; Mitomycins; Picibanil; Tumor Necrosis Factor-alpha

Hepatic resection is generally considered to be superior to any other therapeutic procedures for hepatocellular carcinoma (H.C.C.). However, the resectability of the patients who have HCC. with liver cirrhosis is still low, and surgery is appropriate in only a minority of patients. Although some successful reports of intra-arterial chemotherapy for HCC. have been documented, most of the therapeutic effects are transient and the survival rate is not satisfactory. This report is of a rare case, that of a long-term survivor with HCC treated by intra-arterial chemotherapy and immunotherapy. A 66-year-old man, with a 10-year history of liver cirrhosis was admitted to The Center for Adult Diseases, Osaka, after detection of a tumor in the right lobe on US. On admission, serum AFP was within normal range, HBs-Ag was negative, and ICG-R 15 was 20.8%. On hepatic angiogram, a hypervascular tumor (6 cm in size) was recognized in the middle of the right lobe. He was assessed as unresectable because of insufficient reserve capacity, and the catheterization of the hepatic artery for intra-arterial chemotherapy and the injection 35 KE of OK-432 into the tumor were carried out under laparotomy. After the procedure, the patient was treated by intra-arterial infusion of doxorubicin (ADR) at a total dose of 150 mg and 5-FU in total dose of 25 g, with a hypodermic injection of OK-432 at a total dose of 161 KE. Hepatic angiography, carried out one year after the procedure, disclosed no foci in the liver. The duration of complete remission continued more than 5 years. The patient eventually died of intrahepatic recurrence, but he lived for 7 years and 3 months after the catheterization.

Topics: Aged; Antineoplastic Combined Chemotherapy Protocols; Biological Products; Carcinoma, Hepatocellular; Combined Modality Therapy; Doxorubicin; Fluorouracil; Humans; Infusions, Intra-Arterial; Injections; Injections, Intradermal; Liver Neoplasms; Male; Picibanil; Remission Induction

Topics: Animals; Biological Products; Carboxymethylcellulose Sodium; Carcinoma, Hepatocellular; Catheterization; Dextrans; Drug Combinations; Embolization, Therapeutic; Gelatin Sponge, Absorbable; Hepatic Artery; Humans; Iodamide; Iodized Oil; Iron; Liver Neoplasms; Liver Neoplasms, Experimental; Male; Middle Aged; Picibanil; Rabbits

In order to prevent endotoxemia after hepatectomy in cirrhotic patients, we administered OK-432 before and after hepatectomy to activate the reticuloendothelial function and studied its effect on postoperative endotoxemia. In the cirrhotic group without OK-432 administration (7 patients), the value of endotoxin increased significantly after hepatectomy, compared to the cirrhotic group which received OK-432 administration (5 patients) and the non-cirrhotic group (12 patients), and the endotoxin level was still higher than the preoperative value even on the 14th day. On the other hand, the cirrhotic group with OK-432 administration and the non-cirrhotic group showed minimal increases of endotoxin levels at the first day, which returned to the preoperative values at the third day. Base on these findings, it is suggested that activation of the reticuloendothelial function bears substantial significance as one of the therapeutic modalities for prevention of endotoxemia after hepatectomy in cirrhotic patients.

Topics: Biological Products; Carcinoma, Hepatocellular; Endotoxins; Hepatectomy; Humans; Liver Cirrhosis; Liver Neoplasms; Mononuclear Phagocyte System; Picibanil; Postoperative Complications

Interferon-gamma (IFN-gamma) was induced from a human peripheral mononuclear fraction by incubation with a streptococcal preparation stabilized with penicillin G (OK432). This IFN-gamma-producing activity was significantly reduced in patients with chronic hepatitis and hepatocellular carcinoma. In patients with liver cirrhosis it was also reduced but not significantly. Serum hepatitis B virus DNA and skin tests for the purified protein derivative of tuberculin, phytohemagglutinin-P and a polysaccharide fraction prepared from streptococcus pyogenes Su strain were determined to have no significant relation to this IFN-gamma-producing activity. Although the addition of interleukin 2 (IL-2) to the culture medium enhanced the IFN-gamma-producing activity, there was no difference in this enhancement between normal control and chronic hepatitis. Therefore reduction of the IFN-gamma-producing activity observed in chronic hepatitis seems to be caused by a decreased number of IFN-gamma-producing activity cells or hypofunction of these cells or both. Since HBeAg became negative in patients whose IFN-gamma-producing activity was increased by the administration of the immunopotentiator OK432 or IFN-beta, the IFN-producing system in the patients with B type hepatitis may contribute to the elimination of HBV. Adenine arabinoside suppressed IFN-gamma-producing activity both in vivo and in vitro.

Topics: Adult; Carcinoma, Hepatocellular; Female; Hepatitis; Humans; In Vitro Techniques; Interferon-gamma; Leukocytes; Liver Cirrhosis; Liver Diseases; Liver Neoplasms; Male; Middle Aged; Picibanil

Natural killer (NK) activity in the peripheral blood of patients with chronic liver disease was measured using 51Cr labeled K562 cells as target cells. NK activity was elevated but not significantly in patients with chronic hepatitis compared with healthy controls and significantly lower in the patients with hepatocellular carcinoma. The activity decreased in the order of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. Although the level of NK activity in patients with chronic hepatitis did not correlate with the level of alanine aminotransferase (ALT), it tended to be elevated in association with elevation of ALT in patients treated with OK432, interferon-beta, glycyrrhizin or adenine arabinoside. In chronic liver disease, phytohemagglutinin (PHA) skin test showed a positive correlation with NK activity. In all patients who were treated with the immunopotentiator, OK432, and whose HBeAg became negative, NK activity was elevated during the treatment. These results suggest that the NK activity in peripheral blood may be related to hepatocytic injury even if this is not the effector mechanism of the injury.

Topics: Adult; Carcinoma, Hepatocellular; Chronic Disease; Cytotoxicity, Immunologic; Female; Glycyrrhetinic Acid; Glycyrrhizic Acid; Hepatitis B; Humans; Interferon Type I; Killer Cells, Natural; Liver; Liver Cirrhosis; Liver Neoplasms; Male; Middle Aged; Picibanil; Radioimmunoassay; Skin Tests; Vidarabine

Twenty-one patients with nonresectable hepatocellular carcinoma (HCC) received intraarterial infusion chemotherapy of Adriamycin (Adria Laboratories, Columbus, Ohio) via an indwelling catheter in the hepatic artery. Additional intratumoral injection therapy of OK-432 (50 KE) was administered to ten of these 21 patients. Nine of the ten patients showed a remarkable decrease in lymphocyte count on the first day after therapy. In all of the patients with a decreased lymphocyte count, computed tomograms (CTs) demonstrated evidence of necrosis associated with a rapid decrease in alpha fetoprotein (alpha-FP). Blastogenesis of lymphocytes in peripheral blood induced by phytohemagglutinin (PHA) increased by 3.99 +/- 1.9 (mean +/- SE) times 4 weeks after therapy. On the basis of these results, we concluded that intratumoral injection therapy of OK-432 apparently produced initiation of necrosis in HCC by cell-damaging activity as well as by improvement of cell-mediated immunity.

Topics: alpha-Fetoproteins; B-Lymphocytes; Biological Products; Carcinoma, Hepatocellular; Doxorubicin; Hepatectomy; Hepatic Artery; Humans; Immunotherapy; Leukocyte Count; Liver Neoplasms; Lymphocyte Activation; Picibanil; T-Lymphocytes; Tomography, X-Ray Computed

Topics: Adult; Biological Products; Carcinoma, Hepatocellular; Doxorubicin; Embolization, Therapeutic; Female; Humans; Injections, Intra-Arterial; Iodized Oil; Liver Neoplasms; Male; Middle Aged; Mitomycin; Mitomycins; Picibanil

The studied patient (70 years old, male) had primary hepatoma with rupture of the liver. We resected his left lobe partially and were able to save him. Since a residual tumor was found after surgery, we injected a total of 30 mg of MMC into the SCA 3 times. Under continuous administration of OK-432, the patient survived for 3 years and 6 months in remission. Because the tumor grew gradually, we started a multi-drug immunotherapy (OK-432 5 KE/2 W/intradermal, PSK/3.0 g/day/P.O., SPG/20 mg/2 W/I. M.) which we have advocated. In 6 months the tumor regressed to 1/3 of its original size. The patient is currently in good condition 4 years and 7 months after the operation. This case demonstrated the superiority of multi-drug immunotherapy over single-drug immunotherapy.

Topics: Aged; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Hepatocellular; Humans; Liver Neoplasms; Male; Mitomycin; Mitomycins; Picibanil; Prognosis; Proteoglycans; Sizofiran

Ten patients with hepatocellular carcinoma (HCC) received intra-tumoral injection of OK-432 (6 patients), 99.5% ethanol (2 patients) or both (2 patients). Under ultrasonographic control, a PTC needle (22 G) was inserted percutaneously into the tumor and OK-432, which was prepared with a solution of Su-strain Streptococcus pyogenes A3, or 99.5% ethanol was injected. Patients were injected with OK-432 repeatedly at one-to two-week intervals (up to 5 times) for a total duration of 5 to 15 weeks. The degree of skin test reaction for Streptococcus pyogenes was increased in all patients after the treatment. Over 40% tumor regression was noted in 6 out of 9 patients who received intra-tumoral injection of OK-432. Complete regression was noted in one patient. Before treatment, Interleukin-2 (IL-2) and lymphokine-activated killer (LAK) cell activity in peripheral blood lymphocytes decreased in HCC patients. Two of 6 patients showed markedly increased activity of LAK-cells one week after treatment with OK-432. One other patient had moderately increased LAK-cell activity after treatment with OK-432. No increase in LAK-cell activity was seen in 3 patients who received intra-tumoral injection of ethanol. An especially increased response of LAK-cell activity was seen in patients with small-sized HCC (diameter below 5 cm).

Topics: Administration, Topical; Aged; Biological Products; Carcinoma, Hepatocellular; Ethanol; Female; Humans; Interleukin-2; Liver Neoplasms; Lymphocyte Activation; Male; Middle Aged; Picibanil; Skin Tests; T-Lymphocytes

Topics: Biological Products; Carcinoma, Hepatocellular; Humans; Killer Cells, Natural; Liver Neoplasms; Lymphokines; Picibanil

1-Hexylcarbamoyl-5-fluorouracil (HCFU) was orally administered at doses of 300-600 mg daily for 7-360 days to 27 inoperable cirrhotics with hepatocellular carcinoma. They were simultaneously treated with transcatheter arterial embolization (TAE), and mitomycin C (MMC) injected into the hepatic artery and/or subcutaneous injection of OK-432. Efficient treatment according to the Koyama and Saito (PR and MR) and Karnofsky classification (I -A and I -B) was obtained in 40.7 and 29.6%, respectively, of all the patients; 81.8 and 54.5%, respectively, in the patients treated with TAE and intraarterial MMC injection and 25.0 and 12.5%, respectively, in those given MMC therapy. However, the total doses of HCFU administered (3-216 g) were not related to either efficient or inefficient treatment. Survival rate obtained 1 year following diagnosis of HCC was much better in the patients treated with TAE and/or MMC therapy (greater than 70%) than in those without these treatments (20%). TAE intervals in HCC patients given repeated TAE and intrahepatic MMC injection were slightly prolonged by oral administration of HCFU (more than 12 g). The results suggest that HCFU might be effective for prevention of HCC recurrence following TAE.

Topics: Administration, Oral; Aged; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Hepatocellular; Embolization, Therapeutic; Female; Fluorouracil; Hepatic Artery; Humans; Liver Cirrhosis; Liver Neoplasms; Male; Middle Aged; Mitomycin; Mitomycins; Picibanil; Prognosis

Topics: Adult; Aged; Biological Products; Carcinoma, Hepatocellular; Contrast Media; Female; Humans; Injections; Liver Neoplasms; Male; Middle Aged; Picibanil; Punctures; Ultrasonography

Topics: Biological Products; Carcinoma, Hepatocellular; Humans; Injections; Liver Neoplasms; Picibanil

A low-molecular-weight fraction (M.W. approximately 700) that specifically impairs the induction of type II interferon in mice by purified protein derivative of tuberculin or OK-432 was isolated from the cell-free ascitic fluid of mice bearing Ehrlich ascites carcinoma. Purification was achieved by ultrafiltration and gel filtration. The inhibitory activity of the isolated fraction was 10 times greater than that of the unfractionated starting material in the impairment of type II interferon induction. The significant inhibition was observed even when 0.2 ml of the 10,000-fold dilution of the fraction, which was previously adjusted to 0.25 A unit at 290 nm absorption, was once treated i.p. in normal mice 48 hr before challenge of type II interferon inducers. This fraction was stable to heating at 56 degrees for 60 min. The active component, however, did not affect the in vivo induction of type I interferon by polyriboinosinic-polyribocytidylic acid or tilorone-HCl. In parallel experiments, an identical low-molecular-weight fraction that impairs the type II interferon induction in mice was isolated from the ascitic fluids of rats bearing AH-100B ascites tumor and from a human hepatoma case with advanced cancer metastatic to the peritoneal cavity. However, nontumorous ascitic fluids obtained from adjuvant-stimulated mice and a human liver cirrhosis case did not contain any such inhibitory activity.

Topics: Animals; Ascitic Fluid; Carcinoma, Ehrlich Tumor; Carcinoma, Hepatocellular; Chromatography, Gel; Humans; Interferon Inducers; Liver Neoplasms; Mice; Picibanil; Ultrafiltration