picibanil and Neoplasms

After approval of national health insurance for non-specific immunomodulation such as OK-432 (1978) and PSK (1980), lentinan, SPG, bestatin and dried BCG vaccine have been tried. Including cytokines such as IL-2, IL-12, IFN, TNF or monoclonal antibodies, they have been widely used as biological response modifiers (BRM). Progress in BRM therapy may be broken down into the first 10 years as development, the next 10 years as disappointment and the most recent 5 years as dream-like progress owing to molecular biological techniques. An interdisciplinary approach has been taken by the Japanese Research Society for Surgical Cancer Immunology, founded in 1980, and the Japanese Society of BRM founded in 1988. Many investigations have been performed on issues such as the clinical evaluation or criteria for responder cases, host immunocompetency, post-operative adjuvant immuno-chemotherapy, locoregional immunotherapy, cytokine therapy, adoptive immunotherapy, and tumor specific immunotherapy. Attention has also focused on malignant tumor injury, surgical stress, the advantages or disadvantages of splenectomy, and discussions of the current status and future prospects in the next century for new BRM therapy.

Topics: Antibodies, Monoclonal; BCG Vaccine; Humans; Immunologic Factors; Immunotherapy, Adoptive; Interferons; Lentinan; Neoplasms; Picibanil; Proteoglycans

Topics: Antibodies, Monoclonal; Humans; Immunologic Factors; Mycobacterium bovis; Neoplasms; Picibanil; Proteoglycans

Topics: Acetylmuramyl-Alanyl-Isoglutamine; Adjuvants, Immunologic; Animals; Bryostatins; Growth Inhibitors; Humans; Immunologic Factors; Lactones; Macrolides; Neoplasms; Phosphatidylethanolamines; Picibanil; Polysaccharides

Effector lymphocytes cytotoxic to autologous tumor cells consist of heterogeneous populations. In this communication, we have presented the results of experiments, in which these heterogeneous effector cells were classified based on their surface markers and modes of differentiation depending upon culture conditions. NK cells with a typical phenotype of CD2+ 3-16+ were one of major populations, which could respond to IL-2 and be differentiated into lymphokine activated killer (LAK) cells. LAK cells were also induced from a CD3+4-8- T cell subset by IL-2 stimulation. Effector cells with a phenotype of CD2+ 3-4-8-16-, which may belong to the T cells at early stages of differentiation, could also be induced by stimulation of immunoadjuvants. Finally, it was shown that the effector cells with a typical phenotype of cytotoxic T lymphocytes, CD3+4-8+16-, and a non-MHC-restricted cytotoxicity were induced by IL-2 in the tumor infiltrating lymphocytes.

Topics: Adjuvants, Immunologic; Antigens, Surface; Cytotoxicity, Immunologic; Humans; Interleukin-2; Killer Cells, Natural; Lymphocyte Activation; Lymphocytes; Major Histocompatibility Complex; Neoplasms; Phenotype; Picibanil; T-Lymphocytes; T-Lymphocytes, Cytotoxic

Topics: Adjuvants, Immunologic; Antibodies, Monoclonal; Antigens, Neoplasm; Antineoplastic Agents; BCG Vaccine; Biological Products; Cell Differentiation; Cell Wall Skeleton; Combined Modality Therapy; Cytokines; Glycoproteins; Growth Substances; Humans; Immunization; Immunization, Passive; Immunotherapy; Interferons; Killer Cells, Natural; Lymphokines; Macrophage Activation; Mucoproteins; Neoplasms; Neovascularization, Pathologic; Picibanil; Recombinant Proteins; Tumor Necrosis Factor-alpha

Topics: Animals; Antibodies, Monoclonal; Antibody-Dependent Cell Cytotoxicity; BCG Vaccine; Biological Products; Humans; Immunotherapy; Interleukin-2; Lentinan; Neoplasms; Picibanil; T-Lymphocytes

Wilms' tumor 1 (WT1) peptide-based vaccination has been reported for its potential usefulness in targeting several cancers. The adjuvant drug OK-432 is known to have potent immunomodulation and therapeutic properties when applied in cancer treatment and may, thus, be important to trigger the appropriate immunological response in paediatric patients with a solid tumor that are vaccinated with a WT1 peptide.. Paediatric patients with a solid tumor were vaccinated with a WT1 peptide and OK-432 once every 2 weeks, for a total of seven times.. Of the 24 patients, 18 completed the scheduled vaccinations. Sixteen patients had local skin symptoms and/or fever. In 1 patient, anaphylactic symptoms emerged at the time of the final injection, but these quickly subsided after the treatment. WT1-specific immunological responses were observed in 4 patients (22.2%). WT1 and HLA class I expression were confirmed in 100% and 85% of primary tumors, respectively.. WT1 peptide vaccine therapy combined with OK-432 appears to be relatively safe for children. However further studies in a larger number of patients are necessary to confirm its safety and efficacy.

Topics: Adjuvants, Immunologic; Adolescent; Adult; Cancer Vaccines; Child; Child, Preschool; Feasibility Studies; Female; Humans; Immunity, Innate; Male; Neoplasms; Picibanil; Treatment Outcome; Vaccination; WT1 Proteins; Young Adult

We conducted a phase I clinical trial of a cancer vaccine using a 20-mer NY-ESO-1f peptide (NY-ESO-1 91-110) that includes multiple epitopes recognized by antibodies, and CD4 and CD8 T cells. Ten patients were immunized with 600 μg of NY-ESO-1f peptide mixed with 0.2 KE Picibanil OK-432 and 1.25 ml Montanide ISA-51. Primary end points of the study were safety and immune response. Subcutaneous injection of the NY-ESO-1f peptide vaccine was well tolerated. Vaccine-related adverse events observed were fever (Grade 1), injection-site reaction (Grade 1 or 2) and induration (Grade 2). Vaccination with the NY-ESO-1f peptide resulted in an increase or induction of NY-ESO-1 antibody responses in nine of ten patients. The sera reacted with recombinant NY-ESO-1 whole protein as well as the NY-ESO-1f peptide. An increase in CD4 and CD8 T cell responses was observed in nine of ten patients. Vaccine-induced CD4 and CD8 T cells responded to NY-ESO-1 91-108 in all patients with various HLA types with a less frequent response to neighboring peptides. The findings indicate that the 20-mer NY-ESO-1f peptide includes multiple epitopes recognized by CD4 and CD8 T cells with distinct specificity. Of ten patients, two with lung cancer and one with esophageal cancer showed stable disease. Our study shows that the NY-ESO-1f peptide vaccine was well tolerated and elicited humoral, CD4 and CD8 T cell responses in immunized patients.

Topics: Adult; Aged; Antigens; Antigens, Neoplasm; Cancer Vaccines; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Female; Humans; Immunity, Humoral; Male; Mannitol; Membrane Proteins; Middle Aged; Neoplasms; Oleic Acids; Peptide Fragments; Picibanil; Treatment Outcome; Vaccines, Subunit

Nutritional deficiencies are believed to be instrumental in producing reduced immune responses in a variety of animal species. Malnutrition may result in an increase or a decrease in immune functions, depending upon its degree, and also the timing and severity of the nutritional protein deprivation. Our experimental data suggest that there is a significant impairment of cytotoxic activity against K-562 and of the ability of spleen cells to produce interferon in protein-deprived mice in comparison with control mice. Paradoxically accelerated tumor growth after administration of OK-432 or Lentinan was also noted in protein-deficient tumor-bearing mice. In addition, a clinical randomized study of advanced or recurrent gastric cancer patients treated with MMC and FT(MF) with or without lentinan was performed. We recognized excellent end-point results only in the lentinan-treated patients with normal protein levels, while no effect of this agent was seen in patients with low serum protein levels (below 5.9/dl). Aggressive postoperative chemotherapy for cases with distant lymph node metastasis was performed under active nutritional support without any depression of metabolic and immunological states, resulting in a good 5-year survival rate (36.9%).

Topics: Adjuvants, Immunologic; Animals; Esophageal Neoplasms; Humans; Lentinan; Mice; Mice, Inbred C3H; Neoplasms; Nutrition Disorders; Picibanil; Stomach Neoplasms

Immuno-chemotherapeutic effects on the growth of MM-48 mammary tumor were studied in syngeneic C3H/He mice fed diets containing a low (D), normal (N) or arginine-supplemented (NA) protein content. The serum protein levels were 5.7 g/dl in N-mice and 3.7 g/dl in D-mice, respectively. On the other hand, no difference was seen between the two groups, with regard to intra-tumoral protein concentrations. The natural killer (NK) activity of spleen cells was significantly lower in D-mice than in N-or NA-mice. Augmentation of NK activity was detected following the i.p. injection of OK-432 or LENTINAN, while no augmentation was recognized in D-mice. Interferon production of cultured spleen cells was significantly reduced in D-mice, and significantly increased in NA-mice compared with N-mice. NK activity was markedly augmented at 7 days after bilateral oophorectomy in N-mice. Both NK and IFN titers were significantly reduced following administration of estradiol every 7 days. The growth of MM-48 tumor was inhibited by daily administration of OK-432 or LENTINAN in N-mice. However, the tumor growth was paradoxically accelerated after administration of the same drugs in D-mice. These findings indicated that the nutritional or endocrine environment of cancer-bearing mice plays an important role in the effect of some kinds of BRMs. A clinical randomized study of advanced and recurrent gastric cancer patients treated with MMC and FT (MF) with or without LENTINAN, was then performed. We recognized excellent end-point results only in LENTINAN-administered patients with normal protein levels, while no effect of LENTINAN was seen in patients with low protein levels (below 6.0 g/dl).

Topics: Animals; Combined Modality Therapy; Endocrine Glands; Female; Fluorouracil; Humans; Lentinan; Mammary Neoplasms, Experimental; Mice; Mice, Inbred C3H; Neoplasms; Neoplasms, Experimental; Nutritional Physiological Phenomena; Picibanil; Stomach Neoplasms

Numerous trials using dendritic cell (DC)-based vaccinations for the treatment of cancer are being carried out. However, an improvement of the quality of DC used is highly warranted. We here generated human monocyte-derived dendritic cells using a 3 day protocol and stimulated the cells using a combination of OK432 (Picibanil), TLR7/8 ligand CL097, and reduced amounts of prostaglandin (PG)E

Topics: Antineoplastic Agents; Cell Differentiation; Cell Movement; Cells, Cultured; Cytokines; Dendritic Cells; Dinoprostone; Humans; Immunotherapy; Ligands; Monocytes; Neoplasms; Oxytocics; Picibanil; Toll-Like Receptor 7; Toll-Like Receptor 8

Most dendritic cell (DC)-based vaccines have loaded the DC with defined antigens, but loading with autologos tumor-derived antigens would generate DCs that activate personalized tumor-specific T-cell responses. We hypothesized that DC matured with an optimized combination of reagents and loaded with tumor-derived antigens using a clinically feasible electroporation strategy would induce potent antitumor immunity. We first studied the effects on DC maturation and antigen presentation of the addition of picibanil (OK432) to a combination of zoledronic acid, tumor necrosis factor-α, and prostaglandin E2. Using DC matured with the optimized combination, we tested 2 clinically feasible sources of autologous antigen for electroloading, total tumor mRNA or total tumor lysate, to determine which stimulated more potent antigen-specific T cells in vitro and activated more potent antitumor immunity in vivo. The combination of tumor necrosis factor-α/prostaglandin E2/zoledronic acid/OK432 generated DC with high expression of maturation markers and antigen-specific T-cell stimulatory function in vitro. Mature DC electroloaded with tumor-derived mRNA [mRNA electroporated dendritic cell (EPDC)] induced greater expansion of antigen-specific T cells in vitro than DC electroloaded with tumor lysate (lysate EPDC). In a therapeutic model of MC38-carcinoembryonic antigen colon cancer-bearing mice, vaccination with mRNA EPDC induced the most efficient anti-carcinoembryonic antigen cellular immune response, which significantly suppressed tumor growth. In conclusion, mature DC electroloaded with tumor-derived mRNA are a potent cancer vaccine, especially useful when specific tumor antigens for vaccination have not been identified, allowing autologous tumor, and if unavailable, allogeneic cell lines to be used as an unbiased source of antigen. Our data support clinical testing of this strategy.

Topics: Animals; Antigen Presentation; Cancer Vaccines; Carcinoembryonic Antigen; Cell Differentiation; Cell Proliferation; Dendritic Cells; Dinoprostone; Diphosphonates; Drug Therapy, Combination; HCT116 Cells; Humans; Imidazoles; Immunotherapy, Adoptive; Mice, Inbred C57BL; Neoplasm Transplantation; Neoplasms; Picibanil; Precision Medicine; T-Lymphocytes; Tumor Necrosis Factor-alpha; Zoledronic Acid

Generation of functional dendritic cells (DCs) with boosted immunity after the withdrawal of initial activation/maturation conditions remains a significant challenge. In this study, we investigated the impact of a newly developed maturation cocktail consisting of OK-432 and interferon-gamma (IFN-γ) on the function of human monocyte-derived DCs (MoDCs). We found that OK-432 plus IFN-γ stimulation could induce significantly stronger expression of surface molecules, production of cytokines, as well as migration of DCs compared with OK-432 stimulation alone. Most importantly, DCs matured with OK-432 plus IFN-γ-induced maintained secretion of interleukin-12 (IL-12)p70 in secondary culture after stimulus withdrawal. Functionally, OK-432 plus IFN-γ-conditioned DCs induce remarkable Th1 and Tc1 responses more effectively than OK-432 alone, even more than the use of α-type-1 cytokine cocktail. As a result, DCs matured with OK-432 plus IFN-γ can prime stronger cytotoxic lymphocyte (CTL) and natural killer (NK) cell response against tumor cells in vitro. Peripheral blood mononuclear cells activated by DCs matured with OK-432 plus IFN-γ also showed greater tumor growth inhibition in vivo in null mice. Molecular mechanistic analysis showed that DC maturation using IFN-γ in concert with OK-432 involves the activation of p38 and nuclear factor-kappa B (NF-κB) pathways. This study provided a novel strategy to generate more potent immune segments in DC vaccine.

Topics: Animals; Antineoplastic Agents; Biomarkers; Cell Differentiation; Cell Movement; Culture Media, Serum-Free; Cytokines; Dendritic Cells; Drug Synergism; Female; Humans; Inflammation Mediators; Interferon-gamma; Mice; Mice, Inbred BALB C; Mice, Nude; Monocytes; Neoplasms; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Picibanil; Signal Transduction; T-Lymphocytes, Cytotoxic; Th1 Cells

Cancer vaccines have yet to yield clinical benefit, despite the measurable induction of humoral and cellular immune responses. As immunosuppression by CD4(+) CD25(+) regulatory T (Treg) cells has been linked to the failure of cancer immunotherapy, blocking suppression is therefore critical for successful clinical strategies. Here, we addressed whether a lyophilized preparation of Streptococcus pyogenes (OK-432), which stimulates Toll-like receptors, could overcome Treg-cell suppression of CD4(+) T-cell responses in vitro and in vivo. OK-432 significantly enhanced in vitro proliferation of CD4(+) effector T cells by blocking Treg-cell suppression and this blocking effect depended on IL-12 derived from antigen-presenting cells. Direct administration of OK-432 into tumor-associated exudate fluids resulted in a reduction of the frequency and suppressive function of CD4(+) CD25(+) Foxp3(+) Treg cells. Furthermore, when OK-432 was used as an adjuvant of vaccination with HER2 and NY-ESO-1 for esophageal cancer patients, NY-ESO-1-specific CD4(+) T-cell precursors were activated, and NY-ESO-1-specific CD4(+) T cells were detected within the effector/memory T-cell population. CD4(+) T-cell clones from these patients had high-affinity TCRs and recognized naturally processed NY-ESO-1 protein presented by dendritic cells. OK-432 therefore inhibits Treg-cell function and contributes to the activation of high-avidity tumor antigen-specific naive T-cell precursors.

Topics: Antigen-Presenting Cells; Antigens, Neoplasm; Cancer Vaccines; CD4 Antigens; Exudates and Transudates; Humans; Immunosuppression Therapy; Interleukin-12; Interleukin-2 Receptor alpha Subunit; Membrane Proteins; Neoplasms; Picibanil; Streptococcus pyogenes; T-Lymphocytes, Regulatory

We assessed the efficacy of WT1 peptide pulsed dendritic cell (DC) therapy for various advanced cancers. All patients were vaccinated 5 times for 10 weeks with autologous monocytes derived DC and activated T lymphocytes. We treated a total of 26 patients who had HLA-A2402 or/and HLA-A0201. We evaluated 20 of the 26 patients who finished 5-time vaccination (10 men and 10 women, aged 48-81 years, Mean 64 years) and were diagnosed as follows: 3-pancreas cancer, 2-colorectal, 2-breast, 2-esophageal, 2-lung, 2-uterus, 2-ovarian and 5 others. In Clinical response (RECIST), the result was assessed as CR/PR/SD/PD, 0/7/8/5, respectively. Furthermore, the 7 PRs were resulted from 2-colorectal, and one of each was lung, laryngeal, axis, pancreas and smooth muscle sarcoma cancer. The 4 of 7 PR patients were treated with chemotherapy.

Topics: Aged; Antineoplastic Agents; Cancer Vaccines; Dendritic Cells; Female; Humans; Immunotherapy, Active; Lymphocyte Activation; Male; Middle Aged; Neoplasms; Picibanil; T-Lymphocyte Subsets; WT1 Proteins

In vivo, dendritic cells (DC) are programmed to orchestrate innate and adaptive immunity in response to pathogen-derived "danger" signals. Under particular circumstances, DC can also be directly cytotoxic against tumor cells, potentially allowing them to release tumor associated Ags from dying cells and then prime antitumor immunity against them. In this study, we describe the innate characteristics of DC (OK-DC) generated in vitro after exposure of immature human myeloid-derived DC to OK432, a penicillin-inactivated and lyophilized preparation of Streptococcus pyrogenes. OK-DC produced proinflammatory cytokines, stimulated autologous T cell proliferation and IFN-gamma secretion, expressed CCR7, and migrated in response to MIP-3beta. Moreover, OK-DC displayed strong, specific cytotoxicity toward tumor cell targets. This cytotoxicity was associated with novel, OK432-induced up-regulation of CD40L on the cell surface of OK-DC, and was absolutely dependent on expression of CD40 on the tumor targets. These data demonstrate that maturation of human DC with OK432, an adjuvant suitable for clinical use, induces direct tumor cell killing by DC, and describes a novel CD40/CD40L-mediated mechanism for specific DC antitumor cytotoxicity.

Topics: CD40 Antigens; CD40 Ligand; Cell Death; Cytotoxicity, Immunologic; Dendritic Cells; Humans; Neoplasms; Picibanil; Up-Regulation

The aim of this study was to examine the effect of cytokines on different subsets of NK cells, while especially focusing on CD16(-) CD56(dim) cells and CD16(-) CD56(bright) cells. When human peripheral blood mononuclear cells (PBMC) were cultured with a combination of IL-2, IL-12 and IL-15 for several days, a minor population of CD56(bright) NK cells expanded up to 15%, and also showed potent cytotoxicities against various cancer cells. Sorting experiments revealed that unconventional CD16(-) CD56(+) NK cells (CD16(-) CD56(dim) NK cells and CD16(-) CD56(bright) NK cells, both of which are less than 1% in PBMC) much more vigorously proliferated after cytokine stimulation, whereas predominant CD16(+) CD56(dim) NK cells proliferated poorly. In addition, many of the resting CD16(-) CD56(bright) NK cells developed into CD16(+) CD56(bright) NK cells, and CD16(-) CD56(dim) NK cells developed into CD16(-) CD56(bright) NK cells and also further into CD16(+) CD56(bright) NK cells by the cytokines. CSFE label experiments further substantiated the proliferation capacity of each subset and the developmental process of CD16(+) CD56(bright) NK cells. Both CD16(-) CD56(dim) NK cells and CD16(-) CD56(bright) NK cells produced large amounts of IFN-gamma and Fas-ligands. The CD16(+) CD56(bright) NK cells showed strong cytotoxicities against not only MHC class I (-) but also MHC class I (+) tumours regardless of their expression of CD94/NKG2A presumably because they expressed NKG2D as well as natural cytotoxicity receptors. The proliferation of CD16(+) CD56(bright) NK cells was also induced when PBMC were stimulated with penicillin-treated Streptococcus pyogenes, thus suggesting their role in tumour immunity and bacterial infections.

Topics: Antineoplastic Agents; CD56 Antigen; Cytokines; Cytotoxicity, Immunologic; Fluoresceins; Histocompatibility Antigens Class I; Humans; Killer Cells, Natural; Leukocytes, Mononuclear; Lymphocyte Activation; Neoplasms; Picibanil; Receptors, IgG; Streptococcus pyogenes; T-Lymphocyte Subsets

Protein transduction domains (PTDs) have been used increasingly to deliver reagents to a variety of cell types in vitro and in vivo. We have previously shown that HIV TAT-PTD-containing whole protein antigens (Ags)-transduced dendritic cells (DCs) stimulated Ag-specific CD8+ and CD4+ T cells. Although the cytotoxic T lymphocytes (CTL) activity generated was sufficient to prevent engraftment of mice with Ag-expressing tumors, treatment of tumor-bearing mice with TAT-PTD Ag-transduced DCs resulted in tumor regression in some animals. Recently, several other PTDs were reported to promote higher transduction efficiencies than TAT-PTD. To evaluate the role of individual PTDs in induction of immune responses in tumor vaccination studies, we engineered recombinant fusion Ovalbumin (OVA) that contained three differrent PTDs, including the most efficacious known PTD (polyarginine (R9)-PTD). Our results demonstrated that R9-PTD-containing OVA transduced DCs most efficiently, and that transduction efficacy was closely correlated with the extent of Ag-specific CD4+ and CD8+ T-cell activation in vitro and in vivo. Repeated vaccination with R9-PTD-OVA-transduced DC in (OVA-expressing) tumor-bearing mice induced enhanced antitumor immunity, and elicited complete rejection of tumors when DC was co-injected with adjuvants. This vaccination strategy may be clinically applicable, and offers theoretical and practical advantages to those that are in current use.

Topics: Adjuvants, Immunologic; Animals; Antigen Presentation; Antineoplastic Agents; Cancer Vaccines; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Dendritic Cells; Epitopes; Female; Histocompatibility Antigens Class I; Histocompatibility Antigens Class II; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Neoplasm Transplantation; Neoplasms; Ovalbumin; Peptides; Picibanil; T-Lymphocytes, Cytotoxic

Although we have reported that Toll-like receptor (TLR) 4 is involved in OK-432-induced anti-cancer immunity, its detailed mechanism remained uncertain. We hypothesized that OK-432 may first be captured, dissolved by phagocytes, and then active components released from the cells may stimulate TLR4. This hypothesis was examined by the current in vitro experiments. We used TS-2 MoAb which recognizes OK-PSA, an active component of OK-432. First, we observed that OK-432-induced cytokine production by dendritic cells (DCs) and macrophages was significantly inhibited by cytochalasin B, a phagocytosis inhibitor. Immunofluorescence staining by using TS-2 clearly demonstrated that OK-432 was captured and dissolved by these cells. OK-PSA was detected in the supernatants derived from OK-432-treated DC culture by ELISA with TS-2. The supernatants from OK-432-treated DC culture, but not from untreated DC culture, increased NF-kappaB activity in TLR4-expressing cells. The increased NF-kappaB activity was inhibited by TS-2. These findings elucidated the mechanism mediated by phagocytosis and TLR4 signaling in the OK-432 action.

Topics: Animals; Cytochalasin B; Dendritic Cells; Humans; In Vitro Techniques; Macrophages, Peritoneal; Membrane Glycoproteins; Mice; Neoplasms; Phagocytosis; Picibanil; Receptors, Cell Surface; Signal Transduction; Toll-Like Receptor 4; Toll-Like Receptors

It is important to augment the anti-cancer host response in cancer treatment. Recent studies suggested that the signaling via Toll-like receptors (TLRs) which are newly identified receptor molecules recognizing many pathogens, are involved in the induction of anti-cancer immunity. Seya et al. demonstrated that maturation of dendritic cells (DCs) and cytokine induction by the cell wall skeleton of Mycobacterium bovis bacillus Calmette-Guerin (BCG-CWS) are induced via both TLR2 and TLR4. Akira et al. discovered a new molecule of TLR family, TLR9, recognizing unmethylated bacterial CpG-DNA, whose clinical use is expected for cancer therapy as a potent inducer of a helper T cell 1 (Th1)-type T-cell response. TLR9-deficient mice did not show any responses to CpG-DNA, including Th 1 cytokine production and maturation of DCs. We have obtained two molecules, a lipoteichoic acid-related molecule isolated from streptococcal agent OK-432, and a plant-derived 55-kDa protein that can induce Th1 response and elicit a strong anti-cancer effect in vivo and in vitro. Our basic experiments demonstrate that TLR4 signaling is intimately involved in anti-cancer immunity induced by these immunopotentiators. Our clinical examination in oral cancer patients also suggests the requirement of both TLR4 and MD-2 in the OK-432-induced anti-cancer host response. Establishment and clinical use of the methodology for human cancer therapy by utilizing TLR signaling is greatly expected.

Topics: Adjuvants, Immunologic; Animals; Antigens, Surface; CpG Islands; Cytokines; Dendritic Cells; DNA-Binding Proteins; Drosophila Proteins; Humans; Interferon-gamma; Lymphocyte Antigen 96; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Knockout; Models, Immunological; Mouth Neoplasms; Mycobacterium bovis; Neoplasms; Neoplasms, Experimental; Picibanil; Receptors, Cell Surface; Th1 Cells; Toll-Like Receptor 2; Toll-Like Receptor 4; Toll-Like Receptor 6; Toll-Like Receptor 9; Toll-Like Receptors

We have previously isolated a lipoteichoic acid (LTA)-related molecule (OK-PSA) from OK-432, a streptococcal agent, by affinity chromatography on a CNBr-activated Sepharose 4B bound TS-2 monoclonal antibody (mAb) that neutralizes the interferon (IFN)-gamma-inducing activity of OK-432. In the current study, we compared the cytokine-inducing and anti-tumor activities of OK-PSA, a TS-2-binding fraction, with those of OK-PTF, a TS-2-unbinding fraction, in order to determine the efficacy of OK-PSA for clinical use in affinity chromatography using TS-2. In the in vitro experiments using human peripheral blood mononuclear cells (PBMCs), OK-PSA markedly induced Th1-type cytokines, while interleukin (IL)-6 and IL-10, Th2-type cytokines, were induced by OK-PTF. Th1-cytokine induction by OK-PTF was not dose-dependent and was suppressed when PBMCs were treated with a high concentration of OK-PTF. In a mouse model, Th1 cytokines were also induced by OK-PSA and Th2 cytokines were induced by OK-PTF. Th2 cytokine-inducing activity of OK-PTF was accelerated in tumor-bearing mice relative to that in healthy mice. Although the anti-tumor effect of OK-PTF was statistically significant, it was much weaker than that of OK-PSA. A significant difference between the anti-tumor effect of OK-PSA and that of OK-PTF was observed (P<0.05). Finally, OK-PSA elicited its cytokine-inducing effect via Toll-like receptor (TLR) 4, whereas OK-PTF-induced signaling was mediated by both TLR2 and TLR4. These findings strongly suggested that the affinity chromatography using TS-2 is a useful strategy to separate the effective component for cancer therapy (OK-PSA) from other components.

Topics: Animals; Antibodies, Monoclonal; Antibodies, Neoplasm; Cells, Cultured; Chromatography, Affinity; Culture Media; Cytokines; Female; Humans; Hybridomas; In Vitro Techniques; Interferon Inducers; Interferon-gamma; Luciferases; Male; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Monocytes; Neoplasms; Picibanil; Receptors, Cell Surface; Signal Transduction; Th1 Cells; Th2 Cells; Toll-Like Receptor 2; Toll-Like Receptor 4; Toll-Like Receptors

Dendritic cell (DC) administration appears to be a very promising approach for the immunotherapy of cancer. The results of clinical studies have suggested that the nature and the magnitude of antitumor immune responses are critically affected by DC functions, including production of T helper type 1 (Th1)-inducing cytokines, activation of T cell subsets and natural killer (NK) cells, and migration from peripheral tissues to the T cell area of the draining lymph nodes. Administration of immature DCs could fail to fully stimulate antigen-specific immune responses and might induce tolerance under some conditions. In this study, we developed a method to obtain fully mature DCs, and we compared in detail the DCs thus obtained with those obtained using a maturation stimulus termed monocyte-derived medium (MCM)-mimic, which is a mixture of recombinant cytokines and prostaglandin E(2) (PGE(2)) mimicking the components of monocyte-conditioned medium. Using DCs derived from monocytes of advanced cancer patients in this study, we found that DCs stimulated with OK-432 alone showed phenotypes similar to those of mature DCs induced using MCM-mimic, though with better secretion of IL-6 and IL-12. However, these DCs were found to have poor migratory capacity associated with the marginal expression of CCR7. When OK-432 was combined with PGE(2), the CCR7 expression and migratory capacity of DCs were significantly improved without impairing other immuno-stimulatory functions. These results suggest that stimulation with the combination of OK-432 and PGE(2) could be applicable as an alternative to MCM-mimic in clinical trials which require fully matured DCs to induce Th1-type immune responses against tumor cells even in patients with advanced cancer.

Topics: Adult; Cell Differentiation; Cell Movement; Clinical Trials, Phase I as Topic; Culture Media, Conditioned; Cytokines; Dendritic Cells; Dinoprostone; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Humans; Immunologic Factors; Immunotherapy; Interleukin-12; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Male; Middle Aged; Monocytes; Neoplasms; Phenotype; Picibanil; Receptors, CCR7; Receptors, Chemokine; Th1 Cells

Although dendritic cell (DC)-based cancer-specific immunotherapy is a potent strategy for various types of carcinomas, few clinical studies have yielded optimal antitumor effects. Systemic immunodeficiency is observed in patients with advanced malignant disease. In this study, we explored the ability to induce antitumor immunity of the cultured monocyte-derived DCs from hosts with advanced malignant disease, using a mouse model. We found remarkable dysfunction of DCs from mice with advanced cancer, which exhibited T helper (Th)2-dominant immunity, and subsequent reduced antitumor immune response. On the other hand, we found dramatic restoration of the ability of DCs to induce optimal antitumor immune responses after systemic administration of streptococcal preparation OK-432 to the tumor-bearing mice, which induced Th1-dominant immunity. In therapeutic experiments, intratumoral injections of immature DCs from the OK-432-treated mice, designated OK-DCs, enhanced inhibition of tumor growth compared with injections of immature DCs from mice with advanced malignancies, designated T-DCs (P < 0.05), leading to significant prolongation in overall survival (P < 0.05). In analysis of cell surface antigens, antigen-presenting capability and interleukin 12 production, we showed functional skewing in T-DCs and significant restorations in OK-DCs. More CD8+ tumor-infiltrating lymphocytes were detected in the mice treated with OK-DCs; furthermore, CTL assays showed that intratumoral injection of OK-DCs induced tumor-specific immune response to spleen as great as those of N-DCs. These results suggested that Th1-dominant immunity might play a crucial role in the differentiation of DCs, and OK-432 might be useful for inducing optimal antitumor effects in DC-based immunotherapy in tumor-bearing hosts.

Topics: Animals; Cell Division; Cell Membrane; Cell Survival; Coculture Techniques; Dendritic Cells; Endocytosis; Female; Immunohistochemistry; Immunologic Factors; Immunotherapy; Interferon-gamma; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Neoplasm Transplantation; Neoplasms; Phenotype; Picibanil; T-Lymphocytes, Cytotoxic; Th1 Cells; Time Factors; Tumor Cells, Cultured

We previously reported that the anti-tumour effect of OK-432 is considerably enhanced by its intratumoral injection together with fibrinogen. In the present study, we generated killer T cells by culturing tumour-infiltrating lymphocytes from thyroid cancer patients who had received this local immunotherapy. Phenotypic analysis revealed that the T cells were positive for CD3+, CD4+, Leu8-, CD45RO+ and T-cell receptor (TCR)alpha beta+, as well as showing strong surface expression of HLA-DR, CD25, LFA-1 and ICAM-1. The generated CD4+ T cells secreted interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha, TNF-beta, and interleukin (IL)-6 (but not IL-4), and exhibited a high level of cytolytic activity against several tumour cell lines. The cytolytic activity of these T cells for Daudi cells was inhibited by preincubation with an anti-intercellular adhesion molecule (ICAM)-1 antibody, but not by preincubation with anti-TCR alpha beta, anti-CD2, or anti-LFA-1 antibodies. Pretreatment with anti-ICAM-1 antibody inhibited T-cell cytolytic activity, but not conjugation with target cells. In addition, incubation with immobilised anti-ICAM-1 enhanced the secretion of IFN-gamma by T cells. We conclude that ICAM-1 expressed on the effector cytotoxic CD4+ T lymphocytes delivers regulatory signals that enhance IFN-gamma secretion.

Topics: Adjuvants, Immunologic; Antineoplastic Agents; CD4-Positive T-Lymphocytes; Cell Adhesion; Cytotoxicity, Immunologic; Humans; Immunotherapy; Intercellular Adhesion Molecule-1; Killer Cells, Natural; Lymphocytes, Tumor-Infiltrating; Neoplasms; Picibanil; T-Lymphocytes, Cytotoxic; Thyroid Neoplasms; Tumor Cells, Cultured

Autologous tumor killing (ATK) has been implicated as an important prognostic factor in cancer patients since the ability of blood lymphocytes to kill freshly isolated autologous tumor cells was strongly associated with good prognosis of the patients. The present study was designed to induce or enhance ATK sensitivity of fresh human tumor cells by heat stress. Brief exposure of fresh human tumor cells to elevated temperature increased their susceptibility to lysis by autologous blood lymphocytes in a short-term (51)Cr release assay. In addition, the heat-elevated ATK sensitivity was confirmed by clonogenic assays. An increase in ATK was observed with unstimulated lymphocytes in 42% of the cases and OK432 (streptococcal preparation)-activated lymphocytes in 80% of the cases. Stimulation of blood lymphocytes with autologous, heat-stressed tumor cells and OK432 resulted in an increase in number of gamma delta T cells, which was associated with elevated ATK activity against the stressed tumor cells. At the clonal level, three gamma delta T-cell clones (V gamma 9/V delta 2+) proliferated in response to autologous, heat stressed tumor cells and/or OK432 and exhibited elevated cytotoxicity against the tumor cells. Western blot analysis revealed an increased expression of heat shock protein (HSP) 70 in heat- treated tumor cells. Some of them expressed HSP70 on their surfaces. The elevated cytoxicity against heat-stressed tumor cells was inhibited by treatment of targets with anti-HSP70 monoclonal antibody (mAb) or of effector cells with anti-V delta2 mAb. Reactivity of gamma delta T cells to autologous, heat- stressed tumor cells was also inhibited by anti-HSP70 mAb. These results indicate that exposure to heat of tumor cells induces ATK susceptibility, especially to OK432-activated effector cells, and suggest that gamma delta T cells may be involved in ATK against stressed tumor cells through recognition of HSP70 on the target cells.

Topics: Adult; Aged; Antineoplastic Agents; Coculture Techniques; Cytotoxicity, Immunologic; Female; Hot Temperature; HSP70 Heat-Shock Proteins; Humans; Male; Middle Aged; Neoplasms; Picibanil; Receptors, Antigen, T-Cell, gamma-delta; T-Lymphocytes; Tumor Cells, Cultured

The effect of a local injection with a streptococcal preparation OK432 on the antitumor vaccination with tumor cells was investigated. Natural killer (NK) cells, which were detected by anti-NK1.1 monoclonal antibody (mAb), increased in the peritoneal exudate cells after an intraperitoneal (i.p.) injection with syngeneic B16 melanoma cells. Furthermore, a concurrent i.p. injection with OK432 efficiently sustained the locally infiltrating NK cells. The OK432 treatment also sustained the augmented NK and lymphokine-activated killer activities in the peritoneal exudate cells. This treatment also increased the ability of the locally infiltrating NK cells to produce interferon gamma in response to the tumor cells. In addition, the concurrent i.p. injection with OK432 in combination with the tumor cells enhanced the capacity of the spleen cells to turn into anti-(B16 melanoma) cytotoxic T lymphocytes after in vitro restimulation. This augmenting effect of OK432 was dependent on NK cells. Moreover, the concurrent injection with OK432 at the time of anti-tumor vaccination significantly enhanced the protective immunity against B16 melanoma at the rechallenge. Taken together, these findings indicate that a concurrent local injection with OK432 in combination with tumor cells efficiently augments the antitumor vaccination effect, in part, by sustaining the locally infiltrating activated NK cells.

Topics: Adoptive Transfer; Animals; Exudates and Transudates; Female; Injections, Intraperitoneal; Interferon-gamma; Killer Cells, Lymphokine-Activated; Killer Cells, Natural; Kinetics; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Neoplasms; Peritoneal Cavity; Phenotype; Picibanil; Spleen; Stimulation, Chemical; T-Lymphocytes, Cytotoxic

Soluble tumor necrosis factor receptor (sTNF-R) is known to inhibit patient immunity via specific binding with the TNF molecule. To examine the possible involvement of sTNF-R in cancer immunotherapy, the plasma levels of sTNF-R of both 55 kDa and 75 kDa origins were estimated when TNF was induced in patients with malignancy using both a polysaccharide preparation (Lentinan) and a streptococcal preparation (OK-432). The pretreatment plasma levels of the 55 kDa and 75 kDa sTNF-R were 1.04 +/- 0.53 and 1.06 +/- 0.34 ng/ml (mean +/- SE), respectively. The plasma levels of TNF were undetectable before treatment. The plasma sTNF-R levels peaked 2 h after the administration of OK-432 and followed the same pattern as the TNF levels in plasma. Both TNF and sTNF-R nearly returned to pretreatment levels at 16 h after the induction of TNF. The peak plasma levels of the 55 kDa and 75 kDa sTNF-R were 2.46 +/- 0.95 and 3.03 +/- 0.88 ng/ml, respectively, but they did not correlate with the plasma TNF levels. When peripheral white blood cells were cultured with the addition of lipopolysaccharide in vitro, an elevation of the 72 kDa sTNF-R was detected. Thus, the plasma source of this soluble receptor can at least be partly attributed to the white blood cells. However, the 55 kDa sTNF-R showed little increase in the cultures, and its source remains unknown. We should therefore be aware of the elevation of plasma sTNF-R levels by the induction therapy of TNF for patients with malignancies because of the immunosuppressive effect of sTNF-R.

Topics: Adult; Aged; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunotherapy; Leukocytes; Male; Middle Aged; Neoplasms; Picibanil; Receptors, Tumor Necrosis Factor; Tumor Necrosis Factor-alpha

The possible use of spleen-derived mononuclear cells (SPMC) for the intentional and economical adoptive immunotherapy of cancer patients was studied. SPMC were obtained from spleens resected surgically from patients with gastric cancer or idiopathic thrombocytopenic purpura (ITP). When SPMC were cultured in recombinant interleukin 2 (rIL2), SPMC, in the form of interleukin-activated killer spleen cells (IL-SP) proliferated in six of eight cases. CD8+ lymphocytes were the major expanding cell population in most SPMC cultures and IL-SP showed a significant cytolytic activity against cultured tumor cells during cell proliferation. When cultured with a streptococcal preparation, OK-432, for 24 to 48 h, SPMC showed cytotoxic activity against tumor cells and were expressed as OK-432 activated killer spleen cells (OK-SP). The effects of supernatants from IL-SP and OK-SP on tumor cell growth were also examined. The supernatants from IL-SP and OK-SP significantly inhibited cell growth in 3 and 10 out of 11 cases, respectively, while those from OK-SP showed higher growth inhibitory activity than those from IL-SP. The results of this study indicate the potential of SPMC as effector cells for the adoptive immunotherapy of cancer patients.

Topics: Adult; Aged; Female; Humans; Immunotherapy, Adoptive; Interleukin-2; Killer Cells, Lymphokine-Activated; Killer Cells, Natural; Lymphocyte Subsets; Male; Middle Aged; Neoplasms; Picibanil; Spleen; Stomach Neoplasms

Among several approaches to augment the therapeutic effect of adoptive immunotherapy, we focused the antitumor synergy between transferred killer cells and host's fresh lymphocytes. Immunotherapy models using murine tumors or clinical experiments revealed that preadministration of immunostimulator such as OK-432, followed by chemotherapeutic agents such as cyclophosphamide, can induce host's non-cytotoxic fresh lymphocytes that act synergistically with cultured killer cells against autologous tumor cells. Immuno-chemo-lymphocytotherapy (a sequential treatment with OK-432, chemotherapy and adoptive immunotherapy) is useful to treat the patients with advanced cancer even if the number of transferred lymphocytes is limited.

Topics: Animals; Combined Modality Therapy; Cyclophosphamide; Humans; Immunotherapy, Adoptive; Interleukin-2; Killer Cells, Lymphokine-Activated; Lymphocytes; Mice; Neoplasms; Picibanil

Topics: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Chemotherapy, Adjuvant; Humans; Immunotherapy; Korea; Neoplasms; Picibanil; Stomach Neoplasms

We previously found that activated peritoneal neutrophils adhered to tumor cells and destroyed them in the cancer ascites of patients who had received intraperitoneal (ip) OK-432 injection therapy. Since tight adhesion to the tumor cell is essential for effective neutrophil-mediated tumor cell destruction, we investigated the mechanism of peritoneal neutrophil adhesion to tumor cells, using a microplate adhesion assay. An in vitro study demonstrated that the adherence activity of the peritoneal neutrophils of patients who received OK-432 injection therapy to tumor cells increased greatly compared to that of blood neutrophils. The expression of the adhesion molecules (CD11a,b,c/CD18) of peritoneal neutrophils, which was determined by an immunofluorescence study, was about four times as much in CD11b and twice as much in CD11c and CD18 compared to that in blood neutrophils. In vitro OK-432 stimulation of normal blood neutrophils increased neither the adhesion to PLC nor the CD11b expression. The enhanced adherence activity of peritoneal neutrophils to tumor cells was significantly inhibited by pretreatment of the neutrophils with anti-CD11b and anti-CD18 monoclonal antibodies (mAb), but not by pretreatment with anti CD11a or anti-CD11c mAb. These results indicated that the increased adhesiveness of OK-432-induced peritoneal neutrophils to tumor cells was due to the enhanced expression of CD11b/CD18. We concluded that CD11b/CD18 molecules on OK-432-induced peritoneal neutrophils play a crucial role in the neutrophil adherence activity against tumor cells, and these results are the first demonstration in the field of human neutrophil function.

Topics: Antigens, CD; CD18 Antigens; Cell Adhesion; Fluorescence; Humans; Macrophage-1 Antigen; Neoplasms; Neutrophils; Picibanil; Receptors, Leukocyte-Adhesion

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

The secretory function of the natural killer (NK) cells has been considered to be essential for cytolytic activity against their target cells. In this study, the distribution and spatial relationship of various cell organelles which participate in the secretory process, i.e., Golgi apparatus, vesicles, granules and microtubules, were examined ultrastructurally in non-treated and OK-432-activated rat NK cells bound to the tumor cells, with special reference to the rod-cored vesicles which are the most characteristic structure of the NK cells. Rod-cored vesicles and their closely related structures, "empty" vesicles, were derived from the end portion of the Golgi trans cisternae and became accumulated in the central area which was surrounded by the Golgi apparatus, nucleus and contact surface. Some of the vesicles appeared to be further transported to the contact area along the microtubules extending from the centrioles toward the bound targets. The access of the vesicles to the contact surface occurred at that portion where subplasmalemmal actin lattice was thin. The distribution of the dense granules and multivesicular bodies was similar to that of the vesicles, but the area of their occurrence was a little wider. At the outer aspect of the Golgi apparatus was situated the endoplasmic reticulum from which transitional vesicles came to the inner or cis cisternae of the apparatus. The present observations indicate that the cell organelles of the conjugated NK cells are purposefully arranged in the cytoplasm in such order that the generated rod-cored vesicles and "empty" ones are efficiently directed toward the bound tumor cells.

Topics: Animals; Cytoplasmic Granules; Golgi Apparatus; Killer Cells, Natural; Lymphoma; Microscopy, Electron; Microtubules; Mitochondria; Neoplasms; Organelles; Picibanil; Rats; Rats, Inbred Strains; Tumor Cells, Cultured

Ten patients with malignant pericardial effusion were treated with intrapericardial injection of OK-432 (penicillin-treated and heat-treated lyophilized powder of the substrain of Streptococcus pyogenes A3). After intrapericardial insertion of a catheter, a maximal volume of pericardial fluid was withdrawn with cytologic confirmation of malignancy. Five or 10 Klinische Einheit (KE) (KE is a unit used to express the strength of a preparation) of OK-432 diluted in 20 ml of saline was injected into the pericardial space in seven and three patients, respectively. It was repeated in case of reaccumulation. Seven patients were treated only once and the remaining three required a second treatment. Complete control of pericardial effusion was achieved in all patients for an average of 329 days (range, 54 to 790 days). Fever and chest pain were experienced in six and five patients, respectively, but were controlled with antipyretics. Two of three patients who received 10 KE of OK-432 experienced hypotension that was successfully controlled with vasopressor drugs with or without reaspiration of pericardial fluid. Rapid reactive reaccumulation of the pericardial fluid was thought to be a cause of hypotension. A follow-up computed tomography (CT) scan was performed in seven patients and a thickened pericardium was noticed in five; no patients had constrictive pericarditis. These results suggest that intrapericardial administration of 5 KE of OK-432 is an effective and safe treatment for malignant pericardial effusion.

Topics: Adult; Aged; Female; Humans; Male; Middle Aged; Neoplasms; Pericardial Effusion; Picibanil; Survival Rate; Tomography, X-Ray Computed

Forty-three patients with advanced cancer were evaluated for the changes of absolute counts of peripheral blood lymphocytes and lymphocyte subsets during chemotherapy or chemoimmunotherapy. In 27 patients who did not respond to the therapy, whole lymphocytes, T cells, helper/inducer T cells and NK cells decreased significantly in number. In contrast, they showed little decrease in 16 cases responded to the therapy. In situ immunohistochemical analysis of the tumor infiltrating lymphocytes was performed on the carcinoma tissues obtained from 25 gastric cancer patients. T cell infiltration, in which helper/inducer T cells were predominant over suppressor/cytotoxic T cells, and NK cell infiltration were found in most of the tissues comprised various carcinoma types of histology. Furthermore, an analysis of a gastric cancer patient treated with systemic administration of 3 BRMs and intratumoral injection of OK-432 indicated that infiltration of cytotoxic T cells and NK cells augmented by cytokines such as IFN-gamma produced from activated helper/inducer T cells and NK cells. Therefore, it is suggested that movements of helper/inducer T cells and NK cells relate to the response to chemotherapy and participate in prognosis of advanced cancer patients.

Topics: Adult; Aged; Aged, 80 and over; Antineoplastic Agents; Colonic Neoplasms; Esophageal Neoplasms; Female; Humans; Immunologic Factors; Injections, Intralesional; Interferon-gamma; Killer Cells, Natural; Leukocyte Count; Lymphocytes; Male; Middle Aged; Neoplasms; Picibanil; Remission Induction; Stomach Neoplasms; T-Lymphocytes, Helper-Inducer

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

OK 432, a well-established immunopotentiator, has been recently noticed as one of biological response modifiers (BRM). IFN-gamma, also called immune interferon, is regarded as an important immunoregulator secreted by T-lymphocytes. In the present study, we measured the in vitro production of IFN-gamma in human peripheral mononuclear cells (PMC) induced by OK 432. PMC were isolated from the peripheral blood with the Ficoll-Conray centrifugation technique. The number of cells for culture was adjusted to 1 x 10(6) cells/ml in RPMI-1640 medium supplemented with 10% fetal calf serum. Incubation was performed over 7 days at 37 degrees C in the presence of OK 432 at 0.17KE/ml in microculture plates. IFN-gamma secreted in the supernatants was measured consecutively during the observation period with radioimmunoassay. IFN-gamma production in PMC from healthy subjects was already detectable at 24 hours of culture and elevated gradually with incubation, reaching as much as 94.3 +/- 44.6 u/ml (mean +/- SD) after 7 days of culture. In contrast, the production of IFN-gamma in patients with cancer was severely suppressed as 21.9 +/- 25.4 u/ml after 7 days of culture (p less than 0.01). Furthermore, both surgical and radiation treatments inhibited the production of IFN-gamma in PMC from patients with cancer.

Topics: Biological Products; Cells, Cultured; Female; Humans; Interferon Inducers; Interferon-gamma; Leukocytes, Mononuclear; Male; Neoplasms; Picibanil

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: Aged; Fibrin Fibrinogen Degradation Products; Humans; Leukocytes, Mononuclear; Neoplasms; Picibanil; Tumor Necrosis Factor-alpha

The present study was performed to see if OK-432, a useful anticancer agent with potent biological response modifier activities, induces effects on heart due to its nature as a streptococcal preparation. Examination of electrocardiograms of cancer patients treated with a variety of doses and routes of OK-432 has revealed that none of 266 patients examined showed any significant change in electrocardiograms (ECGs) after OK-432 treatments, regardless of their pretreatment heart conditions (normal or abnormal ECGs). Furthermore, these patients did not develop any detectable antibodies against heart muscle, either in serum or deposited on heart tissue. Experimentally, antibodies reactive with human heart tissue became detectable in the sera of rabbits immunized with streptococci in complete Freund's adjuvant, confirming the previous reports. However, the repeated injections of OK-432 without the adjuvant did not induce any detectable antibodies. These results indicate that OK-432 can be used for treating cancer patients with little fear of possible side effects on the heart.

Topics: Adult; Aged; Aged, 80 and over; Animals; Biological Products; Electrocardiography; Female; Fluorescent Antibody Technique; Heart Diseases; Humans; Male; Middle Aged; Neoplasms; Picibanil; Rabbits; Streptococcus pyogenes

The clinical efficacy of intratumoral (IT) administration of BRM in 157 patients with unresectable or recurrent tumors was investigated, and the infiltration of lymphocyte subsets into tumor tissues after IT administration of BRM was immunohistologically examined, to analyse its action mechanism. BRMs used in this study were OK-432, tumor necrosing factor (TNF), whole peptide glucagon (WPG), interferon (IFN)-alpha, IFN-beta, IFN-gamma and interleukin-2 (IL-2). Among them, one hundred thirty-one patients were evaluable for clinical effects, and the therapeutic response rate (CR + PR) was determined in 11/131 (8.4%). Higher therapeutic responses were found in the patients with esophageal cancer, pancreatic cancer and breast cancer, respectively. As for the relationship between the clinical efficacy of IT administration of BRM and the injected sites, the injection into metastatic lesions was more effective than that into the primary or local recurrence. A increase of lymphocyte infiltration after IT BRM immunotherapy and a variety of lymphocyte subsets in tumor tissue injected with any BRMs were found immunohistologically. These results suggest that IT BRM immunotherapy may be effective for the control of tumor growth locally through host-mediated action.

Topics: Adjuvants, Immunologic; Drug Evaluation; Glucagon; Humans; Immunohistochemistry; Injections; Lymphocytes; Neoplasms; Picibanil; Remission Induction; Tumor Necrosis Factor-alpha

This study was undertaken to examine the blastogenic responses of PBL to Streptococcal preparation OK-432 and its fractions. The results showed that OK-432 and its fractions, Su-PR and PM, have the mitogenic activity against human lymphocytes. The demonstration that PBL which had been cultured with OK-432 expressed IL-2 receptors and the blastogenic responses of PBL to OK-432 were inhibited by anti-IL-2 antibody suggested that IL-2 and IL-2 receptors might play a central role in OK-432-induced proliferation of human lymphocytes.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; B-Lymphocytes; Biological Products; Humans; Immunotherapy; In Vitro Techniques; Lymphocyte Activation; Middle Aged; Mitogens; Neoplasms; Picibanil; Receptors, Interleukin-2; T-Lymphocytes

Biological response modifiers can be divided into 2 groups; 1) immunomodulator (IM) or immunostimulator (IS) and 2) cytokines. Several IM or IS have been used clinically for the treatment of various cancers in combination with various chemotherapeutic agents. They are effective for prolonging the survival time or remission duration of cancer patients. However, no direct effect on cancer of the IM.IS has been proven. Cytokines such as interferons (IFNs) or interleukin-2 (IL-2) are effective against renal cell carcinoma, melanoma, hairy cell leukemia, multiple myeloma and other tumors even when they are used singly. IM.IS exert their anti-cancer effects through a combination of NK cell and macrophage activation or production of IFNs and ILs. Therefore, each effect is not strong enough to show a direct anticancer effect. Cytokines which are produced by recombinant techniques can be used in large doses and have been shown to have direct effects on certain types of cancers. The future approach is to devise the best combination between cytokines, cytokines and IM.IS, and cytokines and chemotherapeutic agents.

Topics: Adjuvants, Immunologic; BCG Vaccine; Cell Wall Skeleton; Humans; Interferons; Interleukin-2; Mucoproteins; Mycolic Acids; Neoplasms; Picibanil

Effect of mitomycin C (MMC) administration on the generation of cytotoxic cells induced by in vitro activation of peripheral blood mononuclear cells (PBM) with OK-432, a bacterial immunopotentiator, was studied in patients with various carcinomas. Following i.v. injection of a single dose of 12 mg/m2 MMC, the ability of PBM to generate OK-432 activated killer cells was markedly increased. Thus, the cytotoxic activity observed 7 days after MMC administration was significantly augmented as compared to that before treatment. Therefore, the ability to generate lymphokine activated killer (LAK) cells was examined, and significantly increased capacity was observed 5 and 7 days after MMC injection. Then, the OK-432 activated killer cell activity significantly correlated with the LAK activity. After treatment, the distribution of lymphocyte subsets exhibited a significant decrease in the percentage of OKT8+ cells. Leu-11+ cells were also reduced. The results appear to indicate that the imbalance in T-cell subsets and the increase in the ability to induce LAK cells may be related to the augmenting effect of MMC administration on the generation of OK-432 activated killer cells in cancer patients.

Topics: Biological Products; Humans; In Vitro Techniques; Injections, Intravenous; Killer Cells, Natural; Leukocytes, Mononuclear; Lymphocytes; Mitomycin; Mitomycins; Neoplasms; Picibanil; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

Our method of adoptive immunotherapy (AIT) using autologous IL-2-cultured lymphocytes differs from so-called LAK therapy in several points. We (1) obtain cultured lymphocytes from effusion lymphocytes (EL) or regional lymph-node lymphocytes (RLNL), when possible, rather than peripheral blood lymphocytes (PBL), (2) use crude IL-2 to induce T cell proliferation and to maintain killer activity, (3) use sonicated autologous tumor extract as antigen (Ag) to stimulate proliferation of cytotoxic T cells, and (4) pretreat the patients with local administration of OK-432 before AIT to induce effector cells that act synergistically with transferred killer cells. Surface marker analysis showed that OKT3, IL-2 receptor, Leu 2+15- cells were elevated while Leu 11a and Leu 3+8+ cells were decreased. Culture of RLNL augmented the expression of Leu 3+8- marker. Both of PBL and RLNL responded to Ag, and their auto-tumor killing activities were augmented in about half of the patients while rarely decrease by the addition of Ag. Response rates of patients with pleural effusion due to breast cancer and those with liver metastasis of breast cancer were 94% and 60%, respectively. Moreover, the survival was prolonged in the treated patients with pleural effusion or gastric cancer patients with peritoneal dissemination.

Topics: Cells, Cultured; Evaluation Studies as Topic; Humans; Immunotherapy, Adoptive; Interleukin-2; Killer Cells, Lymphokine-Activated; Neoplasms; Picibanil

Terminal malignant tumor cases were treated with immunopotentiator OK-432. The absolute numbers of neutrophils and lymphocytes and different lymphocyte subsets were determined, and SU-PS and PPD skin tests were also performed to monitor the immunologic status of each patient. The SU-PS test changed to positive in one-third of the patients 2-6 weeks after the start of therapy. The SU-PS responding patients showed an increase in lymphocytes and Leu II+ cells, and an elevation of the OK T4/T8 ratio 2 weeks later. In all patients from the responding group, the (OKIal+ - Bl+) cells decreased in the peripheral blood immediately after the positive change in the SU-PS test. In the SU-PS nonresponding group, the OKT4+ cells tended to decline with time, while the OKT8+ cells increased. That is, the OKT4/T8 ratio remained low throughout the test period. In the SU-PS responding group, OK-432 therapy prolonged the survival time.

Topics: Adjuvants, Immunologic; Aged; Biological Products; Female; Humans; Leukocyte Count; Lymphocytes; Male; Middle Aged; Neoplasms; Picibanil; Skin Tests; Streptococcus pyogenes; Tuberculin Test

An experiment to gauge the effect of OK-432 on UFT activation under a combined administration of UFT and OK-432 at a therapeutic dose has been pursued clinically by determining the blood and intratumoral 5-FU concentrations after administration of UFT. UFT was administered orally to a group of AH-130-bearing rats at a dose of 12 mg/kg or 8 mg/kg. In another group that was given combined treatment with OK-432, the OK-432 was administered subcutaneously at a dose of 0.1 KE/kg together with UFT. The intratumoral 5-FU concentration at 4 hours after UFT 12 mg/kg administration showed higher values in th single UFT treated group than in the combined treated group, but at 8 hours no significant difference was found between these two groups as the concentrations were 0.133 +/- 0.027 microgram/g and 0.128 +/- 0.021 microgram/g, respectively. Further, no significant differences were noticed in the blood and intratumoral 5-FU concentrations between both groups after administration of UFT at a dose of 8 mg/kg. Clinically, the blood FT-207 and 5-FU concentrations after UFT administration showed no significant difference between the single UFT treated group and the combined OK-432 treated group, nor was there a significant difference in the intratumoral concentration. In summary, it is considered that OK-432 has little effect on UFT activation when UFT and OK-432 are administered in combination.

Topics: Administration, Oral; Animals; Antineoplastic Combined Chemotherapy Protocols; Biological Products; Fluorouracil; Humans; Liver Neoplasms, Experimental; Male; Middle Aged; Neoplasms; Picibanil; Rats; Rats, Inbred Strains; Tegafur; Uracil

Fourty-three patients with pleuritis carcinomatosa have been treated with an intra-thoracic administration in four distinct ways: Adriamycin alone, OK-432 alone, the combination of Adriamycin with OK-432, or the combination of Mitomycin C with OK-432. Judging from an evaluation of the subsequent chest X-rays, the prognosis of the cases which received the combination therapy of the two drugs was significantly better to that of the others. There were no significant differences between cases with primary lung cancer and those with other cancers. As for the side effects after the administration, there were no side effects, such as the trouble the renal function or liver function, other than fever and a decrease in the W.B.C.

Topics: Adjuvants, Immunologic; Aged; Antineoplastic Combined Chemotherapy Protocols; Biological Products; Doxorubicin; Female; Fever; Humans; Injections; Leukopenia; Mitomycin; Mitomycins; Neoplasms; Picibanil; Pleurisy; Prognosis; Thorax

The clinical efficacy of intratumoral (IT) OK-432 immunotherapy in advanced cancer patients was investigated. Furthermore, the infiltration of lymphocyte subsets into the tumor tissues after the IT administration of OK-432 was also immunohistologically examined in order to analyze the mechanism of action of OK-432 immunotherapy. Forty-four patients with advanced cancer were treated with IT OK-432 immunotherapy. Ten KE (1 mg) of OK-432 was given either daily or on every second day and repeated as often as possible, the mean frequency of OK-432 injections being 18.1 +/- 14.5 times, ranging between 5 and 25 administrations. Thirty-one of the 44 patients were evaluable, 3 of whom (9.7 per cent) developed a partial response and 5 (16.1 per cent) a minor response. Intratumoral OK-432 immunotherapy, however, did not necessarily prolong the survival time. Leu 1, 3 and 7 reactive cells infiltrated into the tumor tissues treated by OK-432 injection, more frequently, when compared with cells which had been treated by recombinant TNF injection. Thus, it was suggested that IT OK-432 immunotherapy might be effective for the local control of tumor growth through the host mediated action, and that, in combination with systemic therapy, may enhance the clinical effects and prolong the survival time in advanced cancer patients.

Topics: Adult; Aged; Antibodies, Monoclonal; Biological Products; Humans; Middle Aged; Neoplasms; Picibanil; T-Lymphocytes

The therapeutic effect of OK-432 induced endogenous TNF on tumor bearing mice and cancer patients was investigated. OK-432 (10 KE/mouse) was administered intraperitoneally to Balb/c mice 7 days prior to the transplantation of Meth A cells (1 x 10(6)/mouse) into the abdominal cavity. And at day 1 of tumor inoculation, 1 KE/mouse of OK-432 was administered intraperitoneally. The significant prolongation of life span was observed in these mice. On the basis of these observation, therapeutic effect of endogenous TNF on cancer patients was clinically evaluated. OK-432 was administered intraperitoneally or intrapleurally to cancer patients with peritonitis carcinomatosa or pleuritis carcinomatosa 4 times (10KE each) every other day and 50KE of OK-432 was readministered with the interval of 7 days. An appreciable activity of TNF was detected in peritoneal fluids or pleural effusion, and the significant decreasing of these fluids was observed. It is therefore concluded that these therapeutic approach may well be taken into account in treatment of cancer.

Topics: Aged; Animals; Ascitic Fluid; Biological Products; Breast Neoplasms; Cytotoxicity, Immunologic; Female; Humans; Immunotherapy; L Cells; Lung Neoplasms; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Neoplasms; Picibanil; Sarcoma, Experimental; Stomach Neoplasms; Tumor Necrosis Factor-alpha

The priming effect of endogenous biological response modifiers (BRMs), interferons (IFNs), and interleukin-2 (IL2), and the triggering effect of BRMs of bacterial origin, OK-432 and Corynebacterium parvum, on endogenous production of the tumor necrosis factor (TNF) were investigated in mice. TNF activity in serum was measured by in vitro cytotoxicity assay with L-929 cells as a target. The i.v. injection of OK-432 (3KE per mouse) triggered TNF maximally (mean value: 30 U/ml) after 2 h, with a similar time course to that of triggering by lipopolysaccharide. The priming activities of IFNs and IL2 were examined in the system of TNF-triggering by OK-432. The i.v. injection of recombinant IFN-gamma (rIFN-gamma, 10(4) U per mouse) increased TNF production to 790 U/ml; this priming effect was observed just after its injection, was maximal after 2 to 6 h, and disappeared after 24 h. Other types of interferon, rIFN-alpha A/D(Bgl) (2 X 10(5) U per mouse), rIFN-beta (10(6) U per mouse), and natural IFN-alpha/beta (10(6) U per mouse) showed maximal priming activity 6 h after their injection (200 to 800 U/ml) but no effect just after their injection. Recombinant IL2 (10(6) U per mouse) had priming activity that showed a similar time course to that of interferons other than IFN-gamma (a maximal TNF production: 100 U/ml). The i.v. injection of C. parvum, like OK-432, triggered TNF production at doses of 0.06 and 0.3 mg per mouse 2 h after its injection and the triggered TNF activity was enhanced by rIFN-gamma. These findings suggest that combinations of the above endogenous BRMs as priming agents and OK-432 or C. parvum as a triggering agent could induce endogenous production of TNF even in human cancer patients. In fact, combined administration of rIFN-gamma and OK-432 produced TNF in human cancer patients. The advantage of this method for treatment of human cancer patients is discussed.

Topics: Animals; Biological Products; Cytokines; Interferons; Interleukin-2; Lipopolysaccharides; Macrophages; Male; Mice; Mice, Inbred C3H; Neoplasms; Picibanil; Propionibacterium acnes; Rabbits; Tumor Necrosis Factor-alpha

Topics: Biological Products; Combined Modality Therapy; Humans; Interferons; Neoplasms; Picibanil

This study was carried out with 48 patients received surgery, i.e., 23 stomach cancer, 8 colon cancer, 6 rectal cancer, 9 breast cancer etc. Patients in group A received UFT in combination with OK-432. Each of UFT or OK-432 was given to the patients in groups B or C, respectively. Changes in the skin reaction to Su-PS were measured before and after dosing, and concentrations of Tegafur and 5-FU in serum and tumor tissues were determined after administration. Analysis of the skin reaction to Su-Ps revealed that patients with positive skin reaction before surgery in group A didn't manifest depression due to sensitization by UFT therapy. Although average values of the skin reaction after dosing were slightly lower compared to those before dosing in group B, sensitization was effective. Values of the skin reaction after dosing were significantly (p less than 0.05) high compared to those before dosing in groups A and C. Concentrations of Tegafur and 5-FU in serum reached to the peak 2 hr later and were maintained high enough to expect clinical responses even at 4 hr after administration in groups A and B. Especially there was not a significant difference between groups A and B in tumor tissue levels of 5-FU, and a high effective concentration was obtained. Combination therapy of UFT with OK-432 exhibited no significant interaction between them in adjuvant immuno-chemotherapy, and satisfactory results were expected in clinical cures.

Topics: Adult; Aged; Aged, 80 and over; Antineoplastic Combined Chemotherapy Protocols; Biological Products; Combined Modality Therapy; Humans; Intradermal Tests; Middle Aged; Neoplasms; Picibanil; Polysaccharides, Bacterial; Preoperative Care; Tegafur; Uracil

We treated terminal malignant tumor cases with the immunopotentiator OK-432 and determined the absolute numbers of neutrocytes, lymphocytes, and lymphocyte subpopulations, carrying out the SU-PS and PPD skin tests. After the start of therapy, SU-PS test changed to positive in one-third of the patients. The SU-PS -responder patients showed an increase in lymphocytes and Leu 11A-positive cells, and an elevation of the OK T4/T8 ratio two weeks later. In the SU-PS-nonresponder group, OKT4-positive cells declined, while OKT8-positive cells increased with time. That is, the OKT4/T8 ratio remained low throughout the test period. In the SU-PS-responder group, OK-432 therapy was found to prolong the survival period.

Topics: Aged; Biological Products; Female; Humans; Lymphocytes; Male; Middle Aged; Neoplasms; Picibanil; Polysaccharides, Bacterial; Skin Tests; Streptococcus pyogenes; Tuberculin Test

Topics: Administration, Oral; Aged; Aged, 80 and over; Biological Products; Combined Modality Therapy; Female; Humans; Immunization, Passive; Male; Middle Aged; Neoplasm Metastasis; Neoplasms; Picibanil; Pilot Projects

It is crucial to define the immunological characteristics of peripheral blood mononuclear cells in order to clarify the physiological state of cancer patients. In this study, we examined prostaglandin E2 (PGE2) and interleukin-1 (IL-1) production by plastic-adherent cells stimulated by lipopolysaccharide (LPS). The secretion of PGE2 and IL-1 into media depended on the dose of LPS. Although the addition of silica resulted in suppression of LPS-induced PGE2 production, it caused augmentation of IL-1 production. The effect of BRM therapy on IL-1 production was evaluated in five cancer patients. The results demonstrated that BRM therapy increased the levels of IL-1 at the late assessment point for 3 patients and their quality of life was improved. These findings suggest that production of IL-1 may be used as a monitor for the effectiveness of biotherapy. The association of PGE2 production with host antitumor response was evaluated in IFN-gamma therapy. The results showed that the PGE2 production ratio increased early in the therapy period and declined gradually, whereas the expression of HLA-DR antigen on monocytes and the level of IL-1 increased during the treatment. The exact mechanism by which BRM activates monocytes is unknown. It is possible that a distinct subpopulation of monocytes is responsible for this effect.

Topics: Biological Products; Cell Adhesion; Cells, Cultured; Colonic Neoplasms; Dinoprostone; Humans; Interleukin-1; Lipopolysaccharides; Lung Neoplasms; Macrophages; Monocytes; Neoplasms; Picibanil; Prostaglandins E; Stomach Neoplasms

In vitro overnight exposure to the streptococcal preparation OK432 of blood and tumor-associated lymphocytes enhanced their lytic activity against autologous, freshly isolated tumor cells from malignant pleural effusions of cancer patients. This enhancement was mediated through factors that were distinct from interferon (IFN) and interleukin 2 (IL 2). Treatment with IFN-alpha, -beta, or -gamma, or IL 2 failed to augment autologous tumor killing (ATK) activity, although the treatment enhanced natural killer (NK) activity. Intrapleural (i.pl.) injection of OK432 induced or enhanced ATK and NK activities of effusion large granular lymphocytes (LGL) in patients who showed a reduction or disappearance of effusion tumor cells. No such increase in ATK and NK activities was seen with effusion LGL of patients who had no clinical benefit from i.pl. OK432 therapy. Blood ATK and NK activities were not consistently modified by the therapy. These results indicate that i.pl. administration of OK432 induces an augmentation of ATK activity of effusion LGL, which may be involved in the elimination of effusion tumor cells. The data also suggest that the host immune defense against tumor may be better represented by tumor-associated lymphocytes than by blood lymphocytes, and that it is important to examine the effect of biological response modifiers on ATK activity of tumor-associated effector cells.

Topics: Biological Products; Cytotoxicity, Immunologic; Humans; In Vitro Techniques; Interferons; Interleukin-2; Killer Cells, Natural; Lymphocytes; Neoplasms; Picibanil

Immunotherapy against cancer gained popularity as a treatment modality based on the experimental model data that seemed to indicate that both specific stimulation of the immune response with antigen-bearing tumor cells and nonspecific stimulation with bacteria and other adjuvant-type compounds could enhance the existing immune response of the host and prevent recurrence or delay tumor growth. The recent, tremendous advances in molecular biology have given scientists the capability to clone individual genes and thereby produce huge quantities of highly purified products of the human genome. The role of biologically active substances as important pharmacologic reagents was initially explored in human with purified interferon. A purified recombinant interferon, using genes cloned and introduced into bacteria, was being tested in cancer-patients. The discovery of other potent biologically active substances, that is IL-2, TNF, IL-1, LT, thymic factor and monoclonal antibodies, has made tenable the initiation of clinical trials in cancer-patients.

Topics: Adjuvants, Immunologic; Glycoproteins; Growth Inhibitors; Humans; Interferons; Interleukin-2; Lentinan; Neoplasms; Picibanil; Proteoglycans; Tumor Necrosis Factor-alpha

In experiments performed with tumor cells isolated from surgical specimens and lymphocytes collected at the time of surgery, tumor cells are often lysed by blood lymphocytes. Some of the effectors are present in the high-density T-cell subset. This population has little or no anti-K562 activity. For the auto-tumor lysis of T cells with high density, CD8-positive cells are mainly responsible. OK432 pretreatment of the effectors potentiates the existing lytic function or induces it in cases in which the cells have no such function. In general, the potentiating effect reflects inherent patterns of cytotoxicity. In contrast to the lysis of autologous tumor cells, cells with low but not high density were induced to lyse K562.

Topics: Antigens, Differentiation, T-Lymphocyte; Antigens, Surface; Biological Products; Cytotoxicity, Immunologic; Humans; In Vitro Techniques; Neoplasms; Picibanil; T-Lymphocytes

Administration of several biological response modifiers (BRMs) to mice strongly augmented natural killer (NK) activity of leukocytes isolated from the liver. This augmentation of NK activity was induced by two synthetic molecules (MVE-2 and poly ICLC), by two BRMs of bacterial origin (formalin-fixed Propionibacterium acnes: P. acnes and a streptococcal cell wall preparation designated OK-432), as well as a single injection of human recombinant interleukin-2 (hrIL 2). All of these BRMs augmented NK activity in the liver to a greater degree than in the spleen. In addition, adherent leukocytes (greater than 90% macrophages) isolated from the liver following P. acnes administration also exhibited augmented macrophage-mediated cytotoxicity. This cytotoxicity was characterized as macrophage mediated and distinguished from NK activity, on the basis of adherence purification, kinetics of cytotoxicity, and target cell selectivity. The results demonstrate that a variety of BRMs induce augmented natural immunity in the liver and suggest that such organ-associated immune responses may play an important role in the antimetastatic effects of BRMs.

Topics: Adjuvants, Immunologic; Animals; Carboxymethylcellulose Sodium; Cytotoxicity, Immunologic; Female; Humans; Interleukin-2; Killer Cells, Natural; Liver; Macrophages; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Neoplasms; Picibanil; Poly I-C; Polylysine; Propionibacterium acnes; Pyran Copolymer; Recombinant Proteins; Spleen

The enhancement of antitumor effects by combination of biological response modifiers (BRM) was investigated on the basis of their action mechanisms. Experimentally, significant inhibition of tumor growth by combination treatment with BRM, which eliminated immune suppressive mechanisms and in turn enhanced immune responses, was observed. Furthermore, inhibition of tumor growth was observed under conditions of uniform biorhythm in mice and the effect of modification of biorhythm was enhanced by combination with BRM. Clinically, combination treatment of plasma exchange and LAK-cell adoptive immunotherapy was discussed.

Topics: Animals; Humans; Immunization, Passive; Immunotherapy; Interferon Type I; Interleukin-2; Liver Neoplasms, Experimental; Mice; Mice, Nude; Neoplasms; Periodicity; Picibanil; Plasma Exchange; Polysaccharides; Stomach Neoplasms; Vitamins

Topics: Animals; Biological Products; Combined Modality Therapy; Electrocoagulation; Humans; Interleukin-2; Mice; Mice, Inbred BALB C; Neoplasms; Neoplasms, Experimental; Picibanil

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

We examined whether OK-432, a streptococcal preparation, could induce tumor necrosis factor in cancer patients. OK-432 was administered at a dose of 100 KE intratumorally to 4 advanced gastric cancer patients and 10KE intracavitary to 8 patients with malignant pleuroperitoneal effusion. The cytostatic activity of the sera and malignant effusions was assayed by the growth inhibition of L929 cells. OK-432 induced significant cytostatic activity in the sera and malignant effusions. The activity was partially neutralized by the monoclonal antibody against human recombinant tumor necrosis factor. These data suggest that OK-432 induces tumor necrosis factor in the sera and malignant effusions of cancer patients.

Topics: Biological Products; Cytotoxicity, Immunologic; Glycoproteins; Humans; Neoplasms; Peritoneal Neoplasms; Picibanil; Pleural Neoplasms; Stomach Neoplasms; Tumor Necrosis Factor-alpha

Human malignant cells, obtained from surgical specimens or pleuro-peritoneal exudate in 105 cases, were cultivated. Tumor-specific immunity was estimated by cytotoxicity assay using mixed autologous lymphocyte-tumor cell culture in 27 cases (40 times in all). Relations of the observed specific immunity to various nonspecific immunological parameters and to clinical responses to immunotherapy were examined. Positive reaction was observed in 14 examinations. Eight of 25 digestive tract cancers and 6 of 15 other cancers were positive. Cytotoxic activity was significantly higher in the group of favorable clinical response than in the group of progressive disease. In the group of positive cytotoxicity, incidence of clinical response was significantly higher than in the group of negative cytotoxicity. A significant positive correlation was found between cytotoxicity and T cell ratio. Incidence of positive cytotoxicity tended to be higher in the cases of single OK-432 administration (6/11). Incidence of clinical response was higher in the cases of combined OK-432 and PSK administration group (4/14). In the positive cytotoxicity group, 3 examinations showed progressive disease and 5 showed no change. It has been suggested that a higher incidence of favorable clinical response is to be expected in case of positive cytotoxicity, if some therapy will be combined with the conventional immunotherapy.

Topics: Adjuvants, Immunologic; BCG Vaccine; Cytotoxicity, Immunologic; Humans; Lymphocyte Activation; Neoplasms; Picibanil; Proteoglycans; Receptors, Fc; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

The reaction of skin to 10-micrograms and 20-micrograms quantities of Su-polysaccharide (Su-Ps) were examined in 117 patients with cancer, and the results obtained were as follows. Reaction was a little greater to the 20-micrograms dose than to the 10-micrograms dose, but the two parameters were much correlated with each other. The rates of patients showing positive reactions to each dose were 30.8% for 10-micrograms and 34.2% for 20-micrograms. The reactions were augmented by administration of OK-432. The augmentation of skin reaction by OK-432 was more prominent at 20-micrograms doses than at 10-micrograms doses, therefore appearing to be Su-polysaccharide dose-dependent. The ideal Su-polysaccharide dose for Su-Ps skin reaction thus seemed to be 20-micrograms.

Topics: Breast Neoplasms; Colonic Neoplasms; Female; Humans; Hypersensitivity, Delayed; Intradermal Tests; Male; Neoplasms; Picibanil; Polysaccharides, Bacterial; Skin; Stomach Neoplasms; Streptococcus

Topics: Animals; Biological Products; Humans; Infusions, Intra-Arterial; Neoplasms; Picibanil; Rabbits

A cytotoxic factor was induced by the injection of LPS into the peritoneal fluids of mice which had been previously primed with a streptococcal antitumor preparation, OK-432. No cytotoxic effect on L-929 cells was observed in the peritoneal fluids of mice singly treated with OK-432 or LPS. Various mouse and human tumor cell lines were effectively killed by this peritoneal cytotoxic factor, though normal cell lines were insensitive, which indicates that this factor is not species-specific. The highest level of cytotoxic activity was obtained when LPS was given to mice 5 days after the injection of OK-432. The optimal time for collection of peritoneal fluids for the cytotoxic factor was 2 h following the LPS injection. Interferon activity was found to be negative by the plaque reduction test using L-929 cells with vesicular stomatitis virus. These results suggest that this cytotoxic factor is similar to the tumor necrosis factor (TNF) in the mouse serum.

Topics: Animals; Ascitic Fluid; Biological Products; Cell Division; Cell Line; Cell Survival; Cytotoxins; Female; Humans; Kinetics; Lipopolysaccharides; Mice; Neoplasms; Picibanil; Species Specificity

Mouse spleen cells, either pretreated in vitro with 100 U/ml of OK-432-induced IFN gamma for 18 h or obtained from mice 24 or 48 h after i.v. injection of OK-432(100 micrograms/mouse), were examined for their anti-tumor effect by Winn's neutralization assay against Meth-A tumor cells in BALB/c mice. Spleen cells treated in vitro or obtained in vivo 24 h after i.v. injection clearly neutralized the growth of admixed Meth-A cells. Two booster injections of 200 U of IFN gamma near the tumor site accelerated this neutralizating effect. In order to determine the effector subpopulation, inhibitory spleen cells were treated with either anti-Thy-1 monoclonal antibody plus complement, anti-asialo GM1 serum plus complement or with adherence on plastic plates followed by Sephadex G-10 column treatment. The effector cell activity in Winn assay was lost only after the removal of macrophages through plastic plate adherence and Sephadex G-10 column treatment, but not after anti-Thy-1 or anti-asialo GM1 treatment, with either in vitro- or in vivo-treated spleen-cell populations. The growth of Meth-A cells was inhibited not only by these activated macrophages in Winn's assay, but also by adoptive transfer of OK-432-induced cytotoxic macrophages intralesionally 4 days after the implantation of 1 X 10(6) Meth-A cells. Our evidence suggests that the systemic action of OK-432 can be explained by the effect of induced IFNgamma, through the activation of macrophages.

Topics: Animals; Biological Products; Cell Line; Cells, Cultured; Female; Injections, Intravenous; Interferon Inducers; Interferon-gamma; Macrophage Activation; Macrophages; Male; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Neoplasms; Picibanil; Spleen

As for the non-specific cancer immunotherapy concerned, the effect is limited and local use of immunopotentiators which could collect effector cells around tumor tissues might be the best way of cancer immunotherapy. Intratumoral or intraperitoneal administration of large-dose of OK-432 (100 KE) has been investigated since many years with favorable results. Side effects were minimum with a few days continuing slight fever elevation. From the immunohistological examinations using monoclonal antibodies, participation of killer T cells and NK cells was confirmed after intratumoral administration of large-dose OK-432. On the other hand, after intraperitoneal administration of OK-432, neutrophil leucocytes appeared at first on the 2nd to 4th day and they were followed by lymphocytes on the 4th to 5th day and finally appeared lot of macrophages on the 6th and later days. From the results of in vitro and in vivo experiments, these macrophages seemed to play the leading role in the cytostatic activities after intraperitoneal OK-432 administration. As for the fear of introducing suppressor cells after large-dose OK-432 administration, detailed studies on suppressor activities before and after operation for gastric cancer patients revealed no particular increase of suppressor cell activities after intratumoral and intraperitoneal administration of 100 KE OK-432 as compared with curative resection cases with no OK-432 administration.

Topics: Adjuvants, Immunologic; Administration, Topical; Animals; Biological Products; Humans; Immunotherapy; Injections, Intraperitoneal; Mice; Neoplasms; Picibanil

Cancer patients with marked lymphocytic infiltration in tumor tissue tend to have a better prognosis than those who do not. This tendency is apparent from our study of the relationship between prognosis and lymphocyte infiltration in tumor tissue and the reactions of regional lymph nodes in several malignant diseases. We used the immunofluorescent technique to determine the percentage of T lymphocytes in the infiltrating cells. We also present our results obtained by the immunoperoxidase technique using monoclonal antibodies (OKT and Leu series).

Topics: Humans; Lymphocytes; Neoplasms; Picibanil; Prognosis; T-Lymphocytes

We have studied the effects of extracorporeally induced systemic hyperthermia on cell-mediated immunity in 12 patients. Also, the effect of heating and anti-cancer drugs on ADCC activity 3 Plaque forming method and 51Cr release method) and NK activity (51Cr release method. target: K-562 cells) of normal human lymphocytes was studied in vitro. The following results were obtained: Lymphocytes, T-cells and IgGFcR+ T-cells counts slightly decreased during the hyperthermotherapy in patients. During initiation of the hyperthermotherapy (Rectal temp. 41.6-41.8 degrees), ADCC activity, PHA, and Con-A induced lymphocyte blastogenesis were slightly depressed, while NK activity was slightly enhanced. At the end of the hyperthermia, ADCC activity, lymphocyte blastogenesis and NK activity were extremely depressed. In vitro, 1 hour of heating at more than 40 degrees C extremely depressed ADCC activity. By heating at 42 degrees C, ADCC activity was depressed depending on heating time, while NK activity was slightly enhanced for the first 10 minutes and then depressed depending on heating time. The administration of anti-cancer drugs (MMC, 5-FU, ADM) did not affect NK activity at all during the heating at 42 degrees C. From these results, it is highly suggested that immunopotentiators should be used during extracorporeally induced systemic hyperthermia in order to make up for the depressed cell-mediated immunity.

Topics: Adult; Antibody-Dependent Cell Cytotoxicity; Combined Modality Therapy; Extracorporeal Circulation; Female; Humans; Hyperthermia, Induced; Immunity, Cellular; Killer Cells, Natural; Lymphocyte Activation; Male; Middle Aged; Neoplasms; Picibanil

Topics: Aged; Agglutinins; Biological Products; Female; Humans; Immunotherapy; Injections, Intradermal; Injections, Intramuscular; Leukocyte Count; Lymphocytes; Male; Middle Aged; Neoplasms; Picibanil; Skin Tests

Topics: Administration, Oral; Adult; Aged; Biological Products; Female; Humans; Immunotherapy; Injections, Intradermal; Injections, Intramuscular; Killer Cells, Natural; Male; Middle Aged; Neoplasms; Picibanil

NK activity of patients with cancer against gastric carcinoma cell line KATO-III, lung carcinoma cell line A-549 as well as K-562 was significantly lower than that in the control group. We investigated in vitro effects of immunopotentiator, OK-432, on the effector cells of both healthy donors and cancer patients. NK activity of healthy donors was clearly augmented after treatment of effector cells with OK-432. NK activity of some patients with cancer, however, showed no augmentation after OK-432 treatment. These results might be helpful for monitoring cancer patients from immunological view point.

Topics: Adjuvants, Immunologic; Biological Products; Cells, Cultured; Humans; Killer Cells, Natural; Neoplasms; Picibanil

OK-432, a streptococcal preparation, augmented the natural killing (NK) activity of peripheral blood lymphocytes of normal donors and cancer patients against both NK sensitive and resistant human target cells in vitro. The enhancement of NK activity was evident after 4 h pretreatment and maximum by 16-24 h. The manifestation of OK-432 induced augmentation required active cell metabolism, RNA and protein synthesis but no DNA synthesis of lymphocytes. The supernatants produced by OK-432 stimulated lymphocyte cultures had no enhancing substance nor interferon. Anti-interferon antibodies did not inhibit boosting activity of OK-432. Large granular lymphocytes were involved in both spontaneous and OK-432 induced cytotoxic activity. The proportion of lymphocytes conjugating to target cells was not changed by OK-432. These results suggest that OK-432 augments cytotoxic activity of large granular lymphocytes having ability to recognize target cells independent of interferon induction.

Topics: Adult; Aged; Antimetabolites; Biological Products; Cytotoxicity, Immunologic; Humans; Immunity, Innate; In Vitro Techniques; Interferons; Killer Cells, Natural; Lymphocytes; Middle Aged; Monocytes; Neoplasms; Picibanil; Temperature

Recently, a streptococcal preparation, OK-432 has been used successfully as an immunopotentiator for immunotherapy in patients with malignant tumors in Japan. In this paper, we report that the administration of OK-432 augments the cytotoxic activity of peripheral blood lymphoid cells against a natural killer (NK) cell-sensitive erythroleukemic cell line, K562, in tumor patients. In patients before or after surgery, sufficient amounts of OK-432 strongly augmented the cytotoxic activity within 3 days after the initial administration of OK-432. Thereafter the levels of cytotoxicity declined rapidly. The administration of a lower dose of OK-432 gave a lower increase in cytotoxicity. Enhanced cytotoxicity occurred with the reintroduction of OK-432 but remained at lower levels of activity. Characterization and fractionation of OK-432-induced effector cells revealed that the augmented cytotoxicity seemed to be carried mainly by NK cells. A low titer of interferon was detected in 3 of 10 patients within 72 hr after the first inoculation of the agent. Furthermore, we discuss the potency of OK-432 for the induction of interferon in detail.

Topics: Adjuvants, Immunologic; Biological Products; Breast Neoplasms; Cell Separation; Cytotoxicity, Immunologic; Humans; Interferons; Lymphoma; Neoplasms; Picibanil; Stomach Neoplasms; Time Factors

Topics: Biological Products; Cytotoxicity, Immunologic; Female; Humans; Interferon Inducers; Killer Cells, Natural; Male; Neoplasms; Picibanil

OK-432, a streptococcal preparation, was intradermally injected daily into patients with advanced cancer of the stomach or lung for 4 weeks and the effects of OK-432 on the mitogenic responses of cancer patients were followed. The cells involved in the depression of the response of untreated cancer patients were characterized. The cells responsible for impaired responses were nylon wool non-adherent and suppressed the mitogen responses of autologous and allogeneic lymphocytes. These cells lost their suppressive activity during 7 days' culture in vitro. Following OK-432 immunotherapy, mononuclear cells from cancer patients showed increased responses to PHA and Con A, and nylon wool non-adherent cells did not inhibit the mitogen responses. These results suggest that the cells suppressing non-specific mitogen responses are sensitive to in vitro culture, belong to nylon wool non-adherent cells and are lost during 4 weeks of OK-432 therapy.

Topics: Adult; Aged; Biological Products; Cells, Cultured; Humans; Lung Neoplasms; Middle Aged; Mitogens; Neoplasms; Picibanil; Stomach Neoplasms; T-Lymphocytes, Regulatory

Immunotherapy with daily intradermal injections of OK-432, penicillin- and heat-treated lyophilized powder of Su-strain of streptococcus pyogens A3, for over a period of four weeks resulted in quantitative and qualitative effectiveness on impaired cell-mediated immunity even in many patients with far advanced cancer of the stomach or lung. In vitro lymphocyte studies following immunotherapy with OK-432 demonstrated restoration of circulating lymphocyte counts to more than 1,500/microliters, a level associated with normalized subpopulation constitution and increases of phytomitogen blastogenesis. Furthermore, a delayed hypersensitivity skin reaction to PPD was boosted or converted into a positive reaction in some cases. There was, however, no detectable, definite effect on humoral immunity after the therapy. Survival rates at three and six months after the initiation of immunochemotherapy using OK-432 and another chemotherapeutic agent, 5-fluorouracil, in 40 patients with cancer were significantly longer than those of matched control patients given chemotherapy alone.

Topics: Adult; Aged; Biological Products; Humans; Immunity, Cellular; Immunotherapy; In Vitro Techniques; Leukocyte Count; Lymphocyte Activation; Middle Aged; Mitogens; Neoplasms; Picibanil; Remission, Spontaneous; T-Lymphocytes; Tuberculin Test

Human adherent cells from peripheral blood were cultured with immunostimulant, BCG, yeast wall, or streptococcal preparation (OK-432), for 3 days, and the cytostatic activity of the adherent cells on human tumor cells was examined. The cells cultured in the presence of an immunostimulant exhibited increased phagocytic activity and the number of phagocytosed sheep red blood cells (sRBC) per cell increased. Adherent cells cultured without the immunostimulant showed slight cytostatic activity of 8 approximately 20%. HD-10 cells, derived from Hodgkin's disease, and QG-K and QG-U cells derived from uterine cervical cancer were more susceptible than HeLa cells to the adherent cells activated by OK-432 or yeast cell wall. Relationship between population doubling time and susceptibility to the cytostatic effect mediated by the activated adherent cells was not observed. The supernatant from the activated adherent cells was also effective in inhibiting DNA synthesis of the rapidly proliferating target cells, HD-10 cells and HeLa cells. However, the proliferation of the other two cell lines was enhanced. The effect of activated adherent cells on tumor cell proliferation and its relation to cytostasis were examined and discussed.

Topics: Adjuvants, Immunologic; Cell Adhesion; Cell Line; Cell Wall; Cytotoxicity, Immunologic; Humans; Macrophages; Mycobacterium bovis; Neoplasms; Phagocytosis; Picibanil; Yeasts

Topics: Antineoplastic Agents; Biological Products; Carbazilquinone; Drug Therapy, Combination; Female; Fluorouracil; Humans; Immunotherapy; Mitomycins; Neoplasms; Picibanil

The effect of chemo-immunotherapy on lymphocyte responsiveness to phytohemagglutinin (PHA) and pokeweed mitogen (PWM) was investigated in patients with advanced cancer. Patients received OK-432, a streptococcal preparation, intratumorally and additional systemic MFC (mitomycin-C, 5-fluorouracil, and cytosine arabinoside) chemotherapy intravenously. Patients with high lymphocyte responsiveness to mitogens at the beginning of treatment were more responsive to chemo-immunotherapy than those with low responsiveness. Furthermore, lymphocyte responsiveness to PHA was maintained at a high level throughout the treatment. In contrast, patients with initially depressed lymphocyte responsiveness maintained low responsiveness throughout the treatment. A significant increase of inhibitory activity of the serum was observed in patients with poor results of treatment. From these results, it seems reasonable to conclude that the response to chemo-immunotherapy might be predicted by in-vitro lymphocyte responsiveness.

Topics: Adult; Aged; Biological Products; Female; Fluorouracil; Humans; Immunotherapy; Lymphocyte Activation; Male; Middle Aged; Mitomycins; Neoplasms; Phytohemagglutinins; Picibanil; Pokeweed Mitogens

Topics: Biological Products; Breast Neoplasms; Esophageal Neoplasms; Female; Humans; Lung Neoplasms; Male; Mouth Neoplasms; Neoplasms; Pharyngeal Neoplasms; Picibanil; Radiotherapy Dosage

Topics: Animals; Antineoplastic Agents; BCG Vaccine; Drug Therapy, Combination; Evaluation Studies as Topic; Fluorouracil; Humans; Immunotherapy; Male; Mice; Mitomycins; Neoplasms; Neoplasms, Experimental; Picibanil

Topics: Aged; Biological Products; Drug Therapy, Combination; Female; Fluorouracil; Humans; Intestinal Neoplasms; Lung Neoplasms; Male; Middle Aged; Neoplasms; Pancreatic Neoplasms; Picibanil; Stomach Neoplasms; Tegafur