picibanil and Ascites

Postoperative hepatic lymphorrhea is a very rare complication after abdominal surgery. Hepatic lymphorrhea, not containing chyle, involves an internal lymph fistula between the lymphatic channels toward the cisterna chyli and the peritoneal cavity. Over the past 20 years, 17 cases have been reported in Japan. Here, we report a further case, of a patient with successfully treated intractable hepatic lymphorrhea following gastrectomy for early gastric cancer. We review 18 cases, including the present case, with respect to the management of postoperative lymphorrhea refractory to conventional medical treatment.

Topics: Aged; Ascites; Fistula; Gastrectomy; Humans; Ligation; Liver; Lymph Node Excision; Lymphocele; Male; Picibanil; Postoperative Complications; Recurrence; Sclerotherapy; Stomach Neoplasms; Time Factors

In total, 16 patients with cytologically proven malignant effusion from colorectal cancer were treated by locoregional administration of the streptococcal preparation OK-432 alone or OK-432 plus the T-cell growth factor interleukin (IL)-2, and the action mechanism of the treatment was studied. A positive clinical response, showing a cytologic disappearance of cancer cells and decrease of effusion, was observed in nine of 11 (82%) patients treated with OK-432 alone and in all five patients treated with OK-432 plus IL-2. Flow cytometric analysis revealed that OK-432 plus IL-2 locally induced acute inflammation-like responses, including serial cellular infiltrations of granulocyte migration within a matter of hours, and activation of macrophages and T lymphocyte involvement within the following days, and that a predominant expansion of CD3+CD4+ lymphocytes (CD: cluster of differentiation) was induced by in vitro stimulation with IL-2 of locoregional cells after the OK-432 administration (OK/IL-2AK cells). The OK/IL-2AK cells produced tumour necrosis factor-alpha and interferon-gamma, but these cells did not produce IL-4 and IL-6. The OK/IL-2AK cells expressed potent killing activity against autologous tumour cells. This activity was abrogated by treatment of the lymphocytes with anti-CD3, -CD4, -TCRalphabeta antibody, and by the treatment of target cells with anti-human leukocyte antigen (HLA)-DR antibody. The OK/IL-2AK cells expressed Fas-L gene, and flow cytometric analysis demonstrated HLA-DR expression in approximately 75% of CEA+ or cytokeratin+ effusion cells. TCRVbeta gene analysis of the OK/IL-2AK cells showed an oligoclonal usage of TCRbeta20, which was also involved in the cytotoxic mechanism of the OK/IL-2AK cells. Single-strand conformational polymorphism analysis demonstrated the clonotypes for the TCRVbeta20 gene, and the CDR3s of the gene were sequenced. The clonotypic PCR using the TCRVbeta20-CDR3 sequences could detect the CDR3-identical TCRs in effusion lymphocytes from the other patients. Taken together, it is suggested that locoregional administration of OK-432 plus IL-2 is highly effective for the management of malignant effusion from colorectal cancer. OK-432 plus IL-2 induces autologous tumour-reactive CD4+ Th1 killer lymphocytes, which recognise tumour antigen(s) presented with HLA class II molecules on effusion tumour cells by means of preferential usage of TCRVbeta20. The clonotypic PCR using the TCRVbeta20-CDR3 sequences may be inf

Topics: Adult; Aged; Ascites; CD4-Positive T-Lymphocytes; Colorectal Neoplasms; Female; Humans; Immunotherapy; Interleukin-2; Lymphocyte Activation; Male; Middle Aged; Paracentesis; Picibanil; Pleural Effusion; Reverse Transcriptase Polymerase Chain Reaction

Topics: Adult; Aged; Ascites; Biological Products; Clinical Trials as Topic; Female; Humans; Intestinal Neoplasms; Intestine, Large; Leukocyte Count; Male; Middle Aged; Palliative Care; Picibanil; Prognosis; Skin Tests; Stomach Neoplasms; T-Lymphocytes

Effects of low-dose anti-CD25 antibody on targeting regulatory T (Treg) cells in vitro and in vivo were investigated. Human-mouse chimeric anti-CD25 monoclonal antibody basiliximab was administered into the effusion cavity, followed by locoregional immunotherapy using OK-432 on day 7. Peripheral blood mononuclear cells and effusion lymphocytes (ELs) were collected before and after the basiliximab administration and subjected to further investigations. Surface phenotypes, IFN-gamma production, cytotoxic activity and foxp3 expression of ELs were assessed by flow cytometry, ELISA, 51Cr-releasing assay, and RT-PCR analysis, respectively. We observed that a low concentration of 0.01 microg/ml basiliximab effectively targeted CD4+CD25(bri) Treg cells while preserving CD4+CD25(dim) activated T cells in vitro. This concentration of basiliximab significantly augmented interferon (IFN)-gamma production of ELs when interleukin (IL)-2 was added on day 0 or on day 1 after basiliximab. In the clinical study, intracavitary administration of basiliximab on day 0 followed by OK-432 on day 7 was as safe, well-tolerated, and effective as using OK-432 alone, and a low-dose of 0.002-0.005 mg/kg basiliximab could target CD4+CD25(bri) cells for at least 3 days while relatively preserving CD4+CD25(dim) cells. Foxp3 expression of ELs was not changed definitely by the intracavitary basiliximab. These results suggest that low-dose basiliximab can target Treg cells in vitro and in vivo, and subsequently augment the activation of ELs. Locoregional immunotherapy of malignant effusion using the Treg cell-conditioning regimen with low-dose basiliximab followed by OK-432 administration on day 0 or on day 1 should be evaluated for clinical efficacy in the next phase II trial.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Ascites; Basiliximab; Dose-Response Relationship, Drug; Female; Humans; Interferon-gamma; Interleukin-2; Interleukin-2 Receptor alpha Subunit; Lymphocyte Activation; Male; Middle Aged; Picibanil; Pleural Effusion, Malignant; Recombinant Fusion Proteins; T-Lymphocytes, Regulatory

To evaluate the efficacy of a streptococcal preparation, OK432, on malignant ascites in mice.. PC-C203U (PC203) is a preparation of another strain of the streptococcal family, with the lowest antineoplastic action. To examine the survival curves of mice after the inoculation of BAMC-1 tumor cells, we gave intraperitoneal OK432, PC203, or saline as a control. Intraperitonal neutrophils were counted by cytospin, and interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and macrophage inflammatory protein (MIP)-1alpha were measured by enzyme-linked immunosorbent assay, and 8 h after the administration of OK432, PC203, and saline. Using electron microscopy, we examined the greater omental milky spots, where the in situ proliferation of neutrophils or macrophages takes place.. The OK432 group had the best survival and the control group the worst. The ratio of intraperitoneal neutrophils to BAMC-1 was highest in the OK432 group and lowest in the control group. Quantitative IL-1beta, IL-6 and MIP-1alpha levels were correlated closely with survival. Electron microscopic examination of the milky spots revealed massive proliferation of neutrophils in the OK432 group, but not in the PC203 or control groups.. OK432 effectively activated intraperitoneal neutrophils and a series of immunological chain reactions through an increase in IL-1beta, IL-6, and MIP-1alpha levels. Milky spots could have important antitumor effects in terms of the spread of neutrophils.

Topics: Animals; Ascites; Cell Line, Tumor; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Neutrophils; Peritoneal Neoplasms; Picibanil; Probability; Random Allocation; Reference Values; Sensitivity and Specificity

Locoregional administration of the streptococcal preparation OK-432 is effective in treating malignant ascites from gastric cancer. In order to enhance the efficacy, we conducted a pilot study of locoregional immunotherapy for malignant ascites using host-oriented doses of OK-432. Moreover, action mechanisms of OK-432 were further explored in view of the T-helper type 1 (Th1)-Th2 concept. Gastric cancer patients with cytologically determined malignant ascites were locoregionally administered with OK-432. The dose of OK-432 was selected according to the delayed-type hypersensitivity (DTH) reaction levels to OK-432. Cytokine production profiles of ascites cells were determined using whole ascites assay by stimulation with OK-432. IL-10 mRNA expression was analyzed using RT-PCR. It was found that a positive clinical response was observed in 37 of the 51 (73%) patients with the DTH-oriented approach, showing a significantly higher efficacy than traditional dosage methods using empirical doses (31/58, 53%) (p=0.0487). The DTH-oriented administration of OK-432 produced adverse effects such as fever elevation (p<0.0001) and abdominal pain (p=0.0013) to a significantly lesser extent compared with the traditional treatment. Analysis of the action mechanism of OK-432 revealed that the DTH reaction in responders (19+/-6 mm) was stronger than that in non-responders (6+/-4 mm) (p<0.0001). Tumor necrosis factor (TNF)-alpha production of ascites cells was also higher in responders (3943+/-1247 pg/ml) than in non-responders (1217+/-939 pg/ml) (p=0.0002). There was a significant positive correlation (p=0.0085) between the levels of DTH reaction and TNF-alpha production of ascites cells, but not of blood cells. Responders appeared to polarize on the Th1 axis when clinical responses were plotted on Th1-Th2 dimensions according to the cytokine production profiles of TNF-alpha, IFN-gamma, IL-4 and IL-6 of ascites cells. In vitro culture with IL-2 of ascites cells after OK-432 administration demonstrated an almost clonal expansion of CD4+ lymphocytes, which produced TNF-alpha and IFN-gamma, but did not produce IL-4 or IL-6. IL-10 mRNA expression was detectable in ascites cells from non-responders before treatment. These results suggest that the DTH-oriented locoregional administration of OK-432 may be both effective and less toxic in treating malignant ascites from gastric cancer, showing a possibility of the tailored immunotherapy for malignant ascites. Th1 dysfunction exists

Topics: Aged; Aged, 80 and over; Antineoplastic Agents; Ascites; Female; Humans; Hypersensitivity, Delayed; Immunotherapy; Interleukin-10; Male; Middle Aged; Picibanil; Pilot Projects; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stomach Neoplasms; Th1 Cells; Th2 Cells; Tumor Necrosis Factor-alpha

The case of a patient who developed intractable massive ascites caused by hepatic lymphorrhea derived from surgical injury to the hepatoduodenal ligament is presented. OK-432 was injected intraperitoneally through a catheter advanced to near the hepatoduodenal ligament, so as to expose this site to a high concentration of OK-432. Under ultrasound guidance, it was difficult to reach this site due to massive ascites, and so we performed this procedure under CT and fluoroscopic guidance using a unified CT and angiography system. Subsequently, local administration of OK-432 on five separate occasions resulted in resolution of the ascites. We ascribe this favourable result to the use of this unified CT and angiography system to advance the catheter to the suitable site, making possible the local administration of OK-432.

Topics: Antineoplastic Agents; Ascites; Fluoroscopy; Gastrectomy; Humans; Injections, Intraperitoneal; Male; Middle Aged; Picibanil; Postoperative Complications; Radiography, Interventional; Stomach Neoplasms; Tomography, X-Ray Computed

The intracavitary injection of OK-432 (a streptococcal preparation) with subcutaneous priming has been shown to be an effective immunotherapy for patients with malignant effusion. We applied this treatment in a case of advanced renal cell carcinoma with massive ascites. The patient received 0.2 Klinishe Einheit (KE) OK-432 in the subcutaneous injection twice (day 1 and day 7) followed by 10KE OK-432 intra-abdominal administration (day 9). The treatment was performed safely without major side-effects except for transient pyrexia. A significant reduction of ascites was noted 1 month after the treatment without subsequent re-accumulation. Intracavitary injection of OK-432 with subcutaneous priming seems to be a simple, safe and effective treatment for ascites in advanced renal cell carcinoma.

Topics: Antineoplastic Agents; Ascites; Carcinoma, Renal Cell; Disease Progression; Humans; Injections; Kidney Neoplasms; Male; Middle Aged; Picibanil

Epithelioid haemangioendothelioma is an unusual type of endothelium-derived vascular tumour of borderline malignancy, which has high variability in clinical presentations, depending on the primary site of involvement. We report on a 20-year-old woman who presented with progressive abdominal fullness for 6 months. Multiple lung and liver nodules with pleural effusion and profuse ascites were found. The diagnosis of epithelioid haemangioendothelioma was made after wedge biopsy of the liver. The ascites was intractable and refractory to strong diuretic therapy and repeated paracentesis. Therefore, six courses of intraperitoneal injection of OK-432 were administered. The ascites subsided to a minimal amount after treatment and the patient remained symptom-free for approximately 8 months. The ascites recurred later and another three courses of intraperitoneal injection of OK-432 were administered. The ascites disappeared again. The patient has remained symptom-free since the end of the second period of treatment.

Topics: Adult; Antineoplastic Agents; Ascites; Female; Hemangioendothelioma, Epithelioid; Humans; Injections, Intraperitoneal; Liver Neoplasms; Palliative Care; Picibanil

We have previously reported the development of antitumor effector cells by day 12 after tumor implantation using a murine malignant ascites model with BAMC-1 tumor, which could be cured completely by five consecutive i.p. injections of OK-432 starting on day 2. In contrast, the OK-432 treatment with the same protocol failed to cure the tumor-bearing athymic mice, though it could suppress tumor growth temporarily. The results suggest that T cells may play a critical role in achieving a therapeutic effect. The present study was designed to clarify the nature of the antitumor effector cells induced by OK-432 in euthymic mice. The number of tumor cells in the peritoneal cavity of OK-432-treated euthymic mice increased gradually up to day 12 and dropped suddenly on day 14, while in the athymic mice the tumor cells transiently decreased in the first 7 days then started to expand drastically on day 8. The timing of the appearance of the effector cells was examined by adoptive-transfer experiments. The peritoneal exudate cells (PEC) obtained from BAMC-1 bearing euthymic mice on various days during the treatments with OK-432 were passively transferred intraperitoneally on the respective days (synchronous transfer) or on day 7 (convergent transfer) to BAMC-1-bearing athymic mice, which were treated similarly with OK-432. More than 85% of the recipient athymic mice survived when an adoptive transfer was made on and after day 7. These results indicated that the effector cells developed before day 8 in euthymic mice. The effector cells detectable on day 7 in the PEC represent plastic- or nylon-wool-column-nonadherent cells, which could cure the tumor-bearing athymic mice. Furthermore, the effector cells were destroyed when the nylon-wool-column-nonadherent cells were treated with an anti-L3T4 antibody and complement whereas the same treatment with anti-Lyt2 antibody had no effect. These L3T4+ cells did not possess asialo-GM1 antigen. Although the exact mechanism of action of the effector cells is yet to be clarified, the induction of human equivalents of this type of effector cell would be a good parameter indicative of clinical effects induced by OK-432 or other biological response modifiers in an individual cancer patient.

Topics: Animals; Antigens, Surface; Ascites; CD4-Positive T-Lymphocytes; G(M1) Ganglioside; Immunization, Passive; Immunophenotyping; Immunotherapy; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasms, Experimental; Peritoneal Cavity; Picibanil; Survival Analysis; T-Lymphocytes, Helper-Inducer; Thy-1 Antigens

The antitumor effect of a synthetic lipid A analogue, DT-5461, was investigated using syngeneic tumor models in mice. Intravenous injection of DT-5461 into mice transplanted with solid tumors of MethA fibrosarcoma, MH134 hepatoma, MM46 mammary carcinoma, Lewis lung carcinoma (3LL), and colon adenocarcinomas 26 and 38 resulted in significant reductions in the weight of all tumors except Colon 26, with marked hemorrhagic necrosis of tumor tissues. Efficacy was almost equal to that of an Escherichia coli-type synthetic lipid A (compound 506), and also to those of some chemotherapeutics including Adriamycin, mitomycin C, fluorouracil and cisplatin. Furthermore, DT-5461 was more effective than other immunotherapeutics, including picibanil (OK-432) and lentinan. However, its antitumor effects were inferior to those of Adriamycin or OK-432 against the malignant ascites caused by intraperitoneal inoculation with MethA or with MH134 cells; life span was not prolonged by either intraperitoneal or intravenous administration. In addition, although DT-5461 showed direct inhibitory effects on the in vitro growth of MethA or MH134, these were much weaker than those of Adriamycin. These findings clearly indicated that DT-5461 with systemic administration is a highly effective antitumor agent on solid tumors, and suggest that the antitumor effect of DT-5461 with potent necrotizing activity might derive from indirect mechanisms related to the activation of host immune systems and not to the weak direct cytotoxicity.

Topics: Animals; Antineoplastic Agents; Ascites; Bone Marrow; Disaccharides; Lipid A; Male; Mice; Mice, Inbred Strains; Neoplasms, Experimental; Picibanil; Tumor Cells, Cultured

Although OK-432, a potent BRM, has been known to induce the remarkable improvement of clinical conditions in cancer patients through its strong effects on their immune capabilities, no specific immune parameters have been identified to best predict the clinical outcome after the OK-432 treatment. In an attempt to identify early parameters indicative of the clinical effects, we have administered 0.1 mg of OK-432 intraperitoneally to a total of 12 patients with malignant ascites and examined peritoneal fluid and peripheral blood obtained on 4 days before, 1, 3, and 7 days after the OK-432 injection using various immunobiological assays. Four weeks later, clinical improvements were evaluated by the disappearance of malignant cells from and/or substantial decrease in ascites. Four patients (responders) showed the improvements while 8 patients (nonresponders) showed no clinical evidence for improvement. In a few parameters among the many examined, significantly different patterns of changes were noted between responders and nonresponders. Thus, in nonresponder patients MO and T cell population returned to an initial low level after early increases (on days 1 and/or 3), while they remained increased day 1 through 7 in responders. In responder patients, the cytotoxicity of peritoneal mononuclear cells against K562 and Daudi cells were augmented on day 7, but not in nonresponder patients. The in vitro stimulation of the mononuclear cells with OK-432 enhanced the cytotoxic activity and induced the interferon (IFN) production in the responders but not in nonresponders. These parameters will be useful for the early prediction of the expected clinical effects of OK-432.

Topics: Adult; Aged; Ascites; Cell Survival; Female; Flow Cytometry; Humans; Immunologic Factors; Interferons; Interleukin-1; Interleukin-2; Male; Middle Aged; Monocytes; Neoplasm Proteins; Neoplasms; Picibanil; Tumor Cells, Cultured

Recently, successful treatment of peritonitis carcinomatosa by intraperitoneal administration of lymphokine-activated killer (LAK) cells followed by intraperitoneal recombinant interleukin 2 (rIL-2) has been reported by several authors, in spite of the well documented results of the immunosuppressive activity of malignant ascitic fluid. We investigated the effect of malignant ascitic fluid on in vitro induction of LAK cells obtained from patients' peripheral blood mononuclear cells. We also examined whether Streptococcal preparation OK-432, which has been instilled into the peritoneal cavity to treat malignant ascites, is able to synergize with rIL-2 to induce LAK activity, and the following results were obtained; 1. A little augmentation of LAK activity was observed at a lower concentration of malignant ascitic fluid, while the results were diversified at a concentration higher than 10%. In two cases out of six, a severe suppressive effect was observed at a concentration higher than 40%. At the higher concentration, fewer cells were recovered after a 5 day culture in all cases. 2. No augmentation of LAK activity following the combination of OK-432 with rIL-2 was observed. At a lower concentration of OK-432 (ranging from 0.01-0.04KE/ml), the number of cells increased in comparison with rIL-2 alone. These results suggest that the potential of adoptively transferred LAK cells followed by rIL-2 was not effectively suppressed by malignant ascitic fluid in vivo and that the administration of OK-432 followed by rIL-2 could induce a larger number of various killer cells than rIL-2 alone.

Topics: Ascites; Cytotoxicity, Immunologic; Female; Humans; Interleukin-2; Killer Cells, Lymphokine-Activated; Neutrophils; Ovarian Neoplasms; Picibanil; Tumor Cells, Cultured; Uterine Neoplasms

When a streptococcal preparation, OK-432, was administered intraperitoneally to patients with malignant ascites, lymphocytes with cytotoxic activity against tumor cells increased in number in the peritoneal cavity after 5 to 7 days. To investigate the underlying mechanisms of such lymphocyte accumulation, lymphocyte chemotactic activity (LCA) in ascitic fluid was measured by a modification of the Boyden method. High LCA was found on the third and fourth days after the OK-432 injection. This LCA was generated in the cell-free supernatant of the patients' abdominal neutrophils that accumulated in the peritoneal cavity 24 hours after the injection of OK-432. A similar LCA was also found when normal peripheral neutrophils were incubated with OK-432. Incubation of normal neutrophils without OK-432 failed to generate LCA, however, and OK-432 alone had no LCA. We tentatively named this factor "neutrophil-derived lymphocyte chemotactic factor" (NDLCF). The NDLCF was heat stable and nondializable, and its molecular weight was approximately 45,000 daltons. It attracted mainly natural killer cells by immunoperoxidase assay of migrated lymphocytes in the chemotactic membrane. These characteristics were distinct from C5a, interleukin-1, and interleukin-2. The results suggest that the newly found NDLCF may be responsible for the infiltration of cytotoxic lymphocytes, especially natural killer cells in the peritoneal cavity in patients with malignant ascites when treated by intraperitoneal injections of OK-432.

Topics: Animals; Antibodies, Monoclonal; Antigens, Surface; Ascites; Biological Products; Cells, Cultured; Chemotactic Factors; Chemotaxis, Leukocyte; Humans; Immunoenzyme Techniques; Indomethacin; Injections, Intraperitoneal; Interleukin-1; Interleukin-2; Killer Cells, Natural; Lymphocyte Activation; Mice; Models, Biological; Neoplasms; Neutrophils; Picibanil; T-Lymphocytes; T-Lymphocytes, Cytotoxic

In four ovarian cancer patients with malignant ascites, 10 KE of OK-432 was intraperitoneally administered four times every other day for priming, and 40 KE of OK-432 in a single dose by the same route on day 13 for triggering. The changes in peripheral blood monocytes and intraperitoneal macrophages and the production of tumor necrosis factor (TNF) by peripheral blood mononuclear cells (PBMC) and ascitic lymphoid cells (ALC) were examined. In two of the four patients in whom TNF was induced in the ascites, the TNF production by PBMC and ALC was noted during priming, and after triggering, an increase in both the number of intraperitoneal macrophages and the TNF production by ALC was noted. In two other patients in whom TNF was not detected in the ascites, the ratio of intraperitoneal macrophages to ALC did not change throughout the whole period, and the TNF production by ALC was not augmented. These findings suggest that the priming administration of OK-432 can induce both intraperitoneal macrophages and peripheral blood monocytes into a primed state, and the triggering administration of OK-432 can increase the number of intraperitoneal OK-432-primed macrophages and induce TNF release from these cells.

Topics: Ascites; Biological Products; Cell Count; Female; Humans; In Vitro Techniques; Interferon-gamma; Leukocyte Count; Macrophages; Middle Aged; Monocytes; Ovarian Neoplasms; Picibanil; Tumor Necrosis Factor-alpha

To clarify the action mechanism of OK-432, OK-432 was intraperitoneally (i.p.) administered to 4 ovarian cancer patients with malignant ascites. In 2 of the 4 patients, 4 types of cytokines (tumor necrosis factor, interleukin 1 beta, interleukin 2, and interferon gamma) were induced in ascites after intraperitoneal injection of OK-432 (i.p. OK-432). In one of these patients, the ascitic supernatant after i.p. OK-432 significantly inhibited colony formation of autologous ascitic tumor cells; the amount of ascites decreased markedly, and the ascitic cytology became negative. In the other 2 patients, only interferon gamma was induced in ascites after i.p. OK-432. In these 2, ascitic supernatant did not inhibit colony formation of autologous ascitic tumor cells. These results indicated that i.p. OK-432 can induce various endogenous cytokines in malignant ascites and that these induced cytokines may contribute to the antitumor effect of i.p. OK-432.

Topics: Ascites; Biological Products; Colony-Forming Units Assay; Female; Humans; Interferon-gamma; Interleukin-1; Interleukin-2; Middle Aged; Ovarian Neoplasms; Picibanil; Tumor Necrosis Factor-alpha; Tumor Stem Cell Assay

To four ovarian cancer patients with malignant ascites, 10 KE of OK-432 was intraperitoneally administered four times at 2 day intervals for priming, and 40 KE of OK-432 was given on the 13th day after the first injection for triggering. The changes in blood monocyte and peritoneal macrophage levels and the production of tumor necrosis factor (TNF) by blood mononuclear cells (BMCs) and ascitic lymphoid cells (ALCs) were examined. In the two patients in whom TNF was induced in the ascites, TNF production by BMCs and ALCs was noted during priming. After triggering, increases in both the number of peritoneal macrophages and TNF production by ALCs were noted. In the other two patients, in whom TNF was not detected in the ascites, the ratio of peritoneal macrophages to ALCs did not change throughout the study period, and TNF production by the ALCs was not augmented. These findings suggest that OK-432 can exert a primary effect on both peritoneal macrophages and blood monocytes, and that OK-432 triggering can promote an increase in primed peritoneal macrophages and the release of TNF from these cells.

Topics: Ascites; Biological Products; Female; Humans; Interferon-gamma; Leukocytes, Mononuclear; Lymphocytes; Macrophages; Middle Aged; Ovarian Neoplasms; Picibanil; Polysaccharides; Skin Tests; Tumor Necrosis Factor-alpha

A single injection of a streptococcal preparation, OK-432, with fresh frozen plasma (FFP) (or fresh human serum) into the peritoneal or pleural cavity for the treatment of malignant ascites or pleurisy resulted in a complete reduction of ascitic fluid or pleural effusion in 5 out of 11 patients. FFP was used a further source of complement for the effective accumulation of antitumor polymorphonuclear leukocytes (PMNs) by complement-derived chemotactic factors in the cavity. C5a increased in the fluids 3-9 h after the injection and preceded a massive increase in PMNs. C1 inhibitor (C1INH) and C3b inactivator (C3bINA) decreased in several cases 6 h after the treatment. Chemotactic arachidonic acid metabolites, thromboxane B2(TXB2) as a characteristics of TXA2, and leukotriene B4(LTB4) also increased at the same time even in cases where C5a changed only minimally, and may play a role in accumulating antitumor PMNs in the cavity.

Topics: Adult; Aged; Aged, 80 and over; Ascites; Biological Products; Complement Activation; Complement C5; Complement C5a; Female; Freezing; Humans; Immunization, Passive; Leukotriene B4; Male; Middle Aged; Neoplasms; Neutrophils; Picibanil; Pleurisy; Thromboxane B2

Topics: Ascites; Biological Products; Humans; Male; Middle Aged; Picibanil

Topics: Adjuvants, Immunologic; Ascites; Ascitic Fluid; Biological Products; Female; Humans; Interferon-gamma; Middle Aged; Ovarian Neoplasms; Picibanil; Tumor Necrosis Factor-alpha

Topics: Adult; Aged; Aged, 80 and over; Ascites; Biological Products; Female; Humans; Killer Cells, Natural; Male; Middle Aged; Neutrophils; Picibanil

When a streptococcal preparation, OK-432, was administered intraperitoneally to patients with malignant ascites, the number of neutrophils with cytotoxic activity against tumor cells was increased in the peritoneal cavity immediately after the OK-432 injection. In order to investigate the underlying mechanisms of such neutrophil accumulation, a possible neutrophil chemotactic activity in ascitic fluid was assayed by a modified Boyden method. The chemotactic activity for neutrophils was found significantly higher 6 hr after the OK-432 injection. OK-432 along had no direct chemotactic activity for neutrophils. The chemotactic activity was generated in vitro when ascitic fluid from patients without OK-432 treatment was incubated with OK-432 for 30 min at 37 degrees C. However, preheating of the fluid at 56 degrees C for 30 min or the addition of EDTA to the fluid resulted in the failure of generation of the chemotactic activity after the incubation with OK-432. The addition of EGTA did not show a significant effect. The chemotactic activity in ascitic fluid was found near cytochrome c marker (MW 12,400 D), when fractionated by Sephadex G-200 gel chromatography. The chemotactic activity was heat stable, nondialyzable, and neutralized completely with anti-human complement C5 antibodies. These results suggest that C5a generated via the alternative pathway activated by OK-432 may be responsible for the infiltration of killer neutrophils in the peritoneal cavity in patients with malignant ascites when they are treated by the intraperitoneal injection of OK-432.

Topics: Adult; Aged; Ascites; Biological Products; Chemotaxis, Leukocyte; Complement Activation; Complement C5; Cytotoxicity, Immunologic; Edetic Acid; Egtazic Acid; Hot Temperature; Humans; Middle Aged; Neutrophils; Picibanil

Twelve patients with malignant ascites caused by gastro-intestinal cancer were treated by intraperitoneal administration of OK-432. Tumor cells from these patients were separated from ascitic fluid and cultured in vitro before OK-432 therapy. OK-432 was given intraperitoneally one to two times a week at doses ranging from 5 to 20 KE suspended in saline. Mononuclear lymphocytes were collected from the fluid at various intervals throughout the therapy. The effect of ascites-derived lymphocytes on ascites-derived autologous tumor cell growth was studied in vitro using microplate assay. Nine (responders) of 12 patients showed complete disappearance or significant reduction of ascitic fluid. Ascites-derived lymphocytes slightly inhibited autologous tumor cell growth only in one case before OK-432 therapy. Lymphocytes collected from ascites after OK-432 injection significantly inhibited auto-tumor cell growth in all of 9 responders. In 3 non-responders, however, auto-tumor cell growth inhibition was found only in one case. Interestingly, lymphocytes from non-responders significantly inhibited the growth of tumor cells taken from responders. Conversely, lymphocytes from responders did not inhibit non-responder-derived tumor cell growth. These findings imply that auto-tumor killing by OK-432-induced lymphocytes may depend more on the condition of the tumor cells than on the condition of the lymphocytes, and that the measurement of auto-tumor killing activity by ascites-derived lymphocytes may be useful as an indicator in OK-432 therapy.

Topics: Abdominal Neoplasms; Ascites; Biological Products; Cytotoxicity, Immunologic; Drug Evaluation; Humans; Intestinal Neoplasms; Methods; Peritonitis; Picibanil; Stomach Neoplasms; T-Lymphocytes, Cytotoxic

The role of complement in the polymorphonuclear leukocyte (PMN)-mediated tumor cell destruction in cancer ascites was investigated in relation to a streptococcal preparation OK-432, a so-called biological response modifier. Incubation of OK-432 with fresh human serum at 37 degrees C for 60 min resulted in the generation of C3a and C5a chemotactic factors. Intraperitoneal (i.p.) injection of the mixture to a patient with cancer ascites revealed an accumulation of PMNs in the ascitic fluid for a longer period with a rapid reduction of the ascitic fluid, than an intraperitoneal injection of OK-432 alone examined in the same patient. PMNs were found to invade clusters of the tumor cells and then form rosettes followed by the destruction of tumor cells. These findings induced by OK-432 continued over 10 days in the presence of fresh serum, while diminished within 3-4 days when OK-432 alone was injected. When fresh human plasma or fresh frozen plasma was used instead of serum and i.p. injected with OK-432 avoiding preincubation, the same cytological and clinical changes were observed in other patients. These data strongly indicate that OK-432 activates human complement either in vitro or in the peritoneal cavity, and induces PMNs to accumulate in the ascitic fluid. Although the mechanism of killing of tumor cells by PMNs is obscure, addition of human serum or plasma to i.p. use of OK-432 seems to be valuable for the management of patients with malignant ascites.

Topics: Ascites; Biological Products; Chemotactic Factors; Complement System Proteins; Humans; Interleukin-8; Neoplasms; Neutrophils; Picibanil

Twelve patients with advanced ovarian cancer with ascites were treated by intraperitoneal immunotherapy with OK-432 in combination with surgery and systemic immunochemotherapy. The median time for total resolution of ascites after the OK-432 injection was 7.1 days. In three of the four patients who died, it took more than the average number of days for the ascites to disappear. The ascites index, the ratio of the ascitic fluid volumes before and after intraperitoneal immunotherapy, was significantly higher in the patients who died than in those who survived. The side effects of intraperitoneal administration of OK-432 varied, but were not serious and relief was not needed in most cases. Intraperitoneal injection of OK-432 was considered effective in preventing the reappearance of ascites in these cases of advanced ovarian cancer, and the ascites index was believed to be related to the prognosis.

Topics: Adult; Aged; Ascites; Biological Products; Combined Modality Therapy; Female; Humans; Injections, Intraperitoneal; Middle Aged; Ovarian Neoplasms; Picibanil; Prognosis

The effects of intraperitoneal administration of OK-432 on tumor cells in ascites, in relation to the infiltration of effector cells and on the immune responses of the host, particularly, with regard to immune suppressive mechanisms, were investigated in 25 patients with cancerous ascites. The effects of OK-432 depended on frequency of the repeated and continuous administrations through a tube placed in the peritoneum during laparotomy. Infiltrations of neutrophils and lymphocytes were observed in the ascites within a short period after the administration and monocyte infiltration followed. Disappearance of tumor cells correlated well with the infiltration of these cells. No marked changes in the proliferative responses of peripheral blood lymphocytes were noted and decreases in serum inhibitory factor levels in sera were observed in patients given larger doses of OK-432. A marked reduction in Concanavalin-A-induced suppressor cell activities was observed after OK-432 administration. OK-432 administration probably leads to a disappearance of tumor cells by enhancing peritoneal effector cell activities and by inhibiting the induction of suppressor cell activities, in a dose dependent manner.

Topics: Ascites; Biological Products; Digestive System Neoplasms; Humans; Injections, Intraperitoneal; Killer Cells, Natural; Lymphocyte Activation; Lymphocytes; Picibanil; Stomach Neoplasms; T-Lymphocytes, Regulatory; Time Factors

Topics: Ascites; Biological Products; Humans; Picibanil; Stomach Neoplasms; Streptococcus pyogenes

Twelve patients with malignant ascites caused by gastric cancer were treated with intraperitoneal injections of a streptococcal preparation, OK-432. All had resolution of the ascites after OK-432 treatment. Neutrophils, macrophages, and lymphocytes increased in number in ascitic fluid samples. Some of the OK-432-induced inflammatory cells were attached to tumor cells. The absolute number of tumor cells decreased as the number of infiltrating inflammatory cells increased. Infiltrating lymphocytes were mainly E rosette-forming cells. Infiltrating macrophages were in an activated state. The infiltrating neutrophils, lymphocytes, and macrophages could inhibit DNA synthesis of the patient's own tumor cells in the ascitic fluid after OK-432 injection, but not before the injection. These results indicate that OK-432-induced neutrophils, lymphocytes, probably T cells, and activated macrophages may play an important role in tumor cell destruction in ascites. Moreover, as the number of tumor cells decreased, the ascitic fluid protein levels decreased. Decrease of the ascitic fluid protein level may suppress further accumulation of ascitic fluid, and the low protein level in ascitic fluid is likely to facilitate the reabsorption of the fluid into the bloodstream.

Topics: Adenocarcinoma; Aged; Ascites; Biological Products; Chemotaxis, Leukocyte; Female; Humans; Immunity, Cellular; Leukocyte Count; Macrophages; Male; Middle Aged; Neoplasm Recurrence, Local; Neutrophils; Picibanil; Rosette Formation; Stomach Neoplasms; T-Lymphocytes

Intraperitoneal injections of OK-432, originated from Group A streptococcus pyogens of human origin, were administered to 77 patients with ascites caused by cancer of the digestive tract. Complete disappearance of effusion was observed in 43 cases, its reduction in five out of 77. They received cytologic examination of ascites daily before and after OK-432 injection. One of these patients, a 77-year-old man with carcinomatous peritonitis due to gastric cancer, showed an interesting phenomenon after OK-432 injection. All adenocarcinoma cells in his ascites disappeared at least with 36 hours after OK-432 injection with increasing number of intraperitoneal neutrophils. In addition, neutrophils collected from his ascites or peripheral blood showed cytostatic effect on his ascites-derived tumor cells in vitro. His neutrophil-depleted intraperitoneal cells, however, had no significant effect on DNA synthesis of his tumor cells in vitro. OK-432 itself had no significant effect on DNA synthesis of his tumor cells in vitro. This report describes a patient in whom OK-432-induced neutrophils may play an important role in his tumor cell destruction in ascites.

Topics: Adenocarcinoma; Aged; Ascites; Ascitic Fluid; Biological Products; Cytotoxicity, Immunologic; DNA, Neoplasm; Gastrointestinal Neoplasms; Humans; Male; Neutrophils; Picibanil

Topics: Adult; Aged; Ascites; Biological Products; Female; Gastrointestinal Neoplasms; Humans; Injections, Intraperitoneal; Male; Middle Aged; Picibanil

Topics: Adult; Aged; Ascites; Biological Products; DNA; Female; Humans; Injections, Intraperitoneal; Lymphocytes; Male; Middle Aged; Picibanil; Rosette Formation; Stomach Neoplasms