picibanil and Mast-Cell-Sarcoma

The effect of local injections with streptococcal preparation OK432 on the antitumor effect induced by adoptive immunotherapy (AIT) was investigated. Draining lymph node cells on day 14 after B7-P815 inoculation were used for AIT after in vitro stimulation. AIT on days 7 and 10 showed no effect on the growth of s.c. established P815 mastocytoma, but local injections with OK432 into the tumor sites on days 3, 6 and 9 resulted in a moderate antitumor effect. On the other hand, the combination therapy significantly suppressed tumor growth, and the tumor-bearing mice survived longer than those receiving only one of the treatment modalities. The significant infiltration of CD4+ or CD8+ T cells and multiple necrosis in the tumor sites were observed only when the tumor-bearing mice were treated with the combination therapy. In addition, a transfer experiment using labeled effector cells revealed these infiltrated CD8+ T cells and CD4 T cells to be derived from the donor and the host respectively. More importantly, the combination therapy clearly led to higher expression of the mRNA for Th1-type cytokines and CXC3 chemokines in the tumor sites than resulted from each of the treatment modalities alone. Collectively, these results indicate that local injections with OK432 can help the infiltration of adoptively transferred CD8+ T cells into the tumor sites and synergistically induce the local production of the Th1-type cytokines and CXC3 chemokines.

Topics: Animals; Antineoplastic Agents; B7-1 Antigen; CD8-Positive T-Lymphocytes; Chemokine CXCL10; Chemokine CXCL9; Chemokines, CXC; Combined Modality Therapy; Cytokines; Cytotoxicity, Immunologic; Female; Humans; Immunotherapy, Adoptive; Injections, Intralesional; Intercellular Signaling Peptides and Proteins; Interferon-gamma; Interleukin-12; Lymph Nodes; Lymphocytes, Tumor-Infiltrating; Mast-Cell Sarcoma; Mice; Mice, Inbred DBA; Picibanil; Th1 Cells; Transfection; Tumor Necrosis Factor-alpha

In order to enhance the antitumor vaccination effect of dendritic cells (DC) pulsed with class I tumor peptide, we tried to utilize the local cytokine help of CD4+ T cells reactive to a streptococcal preparation OK432. DC were prepared from murine bone marrow cells by culture with both granulocyte/macrophage-colony-stimulating factor and interleukin(IL)-4. The peritumoral injections of OK432 induced OK432-reactive CD4+ T cells in the draining lymph nodes, and their in vitro production of interferon gamma was thus significantly enhanced by restimulation with OK432-pulsed DC. In addition, anti-P815 mastocytoma cytotoxic T lymphocytes were generated from the in vivo OK432-treated P815-draining lymph node cells only when the lymph node cells were restimulated in vitro with the DC pulsed with both P1A peptide and OK432. Moreover, the peritumoral injections of OK432 and the subsequent vaccination of the DC, pulsed with both OK432 and P1A peptide, significantly suppressed the growth of s.c. inoculated P815. Interestingly, a significant level of IL-12 was detected in the coculture supernatant of both OK432-pulsed DC and OK432-reactive CD4+ T cells. Collectively, our results suggest that the antitumor vaccination effect of DC pulsed with class I tumor peptide could thus be effectively augmented by locally utilizing the Th1-type cytokines from OK432-reactive CD4+ T cells.

Topics: Animals; Antigen Presentation; Antigens, Neoplasm; Bone Marrow Cells; Cancer Vaccines; CD4-Positive T-Lymphocytes; Cell Communication; Cytokines; Dendritic Cells; Female; Immunophenotyping; Interferon-gamma; Interleukin-12; Lymphocyte Activation; Mast-Cell Sarcoma; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Picibanil; T-Lymphocytes, Cytotoxic; Th1 Cells; Tumor Cells, Cultured; Vaccination

Combined effect of cis-DDP (II) (CDDP), a streptococcal preparation (OK-432) and systemic hyperthermia on ascites tumor in mice was studied. Tumor cells were of a syngeneic cloned cell line FMA3 which was derived from Furth's mastocytoma. When number of cells transplanted into the abdominal cavity was at 10(5), a single i.p. injection of CDDP at a dose of 4 mg/kg was effective to increase the mean survival time by a factor of 1.6. OK-432 given i.p. at a dose of 5 KE/kg on every two days between day 2 and day 10 after transplantation decreased the mean survival times. Even when it was used in combination with CDDP, enhancement of the effect of CDDP was not statistically significant. However, by the factorial analysis of the all data obtained in the experiments carried out with tumor cells of the number of 10(2), 10(3), 10(4) and 10(5), enhancement of the effect of CDDP by OK-432 was significant. Systemic hyperthermia of the core-body temperature of 40 degrees C was not effective when used alone, and did not enhance the effect of CDDP. However, combined use of it and OK-432 significantly enhanced the effect of CDDP.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Cisplatin; Combined Modality Therapy; Hyperthermia, Induced; Male; Mast-Cell Sarcoma; Mice; Mice, Inbred Strains; Peritoneal Neoplasms; Picibanil

In vivo tumor inhibitory activity of polymorphonuclear leukocytes (PMN) treated in vitro with lymphokine(s) (LK) was investigated with Winn's assay. Culture supernatants of BALB/c mouse spleen cells incubated with a streptococcal preparation, OK-432, were used as an LK source. With the use of a [3H]uridine release assay, RL male-1 tumor cells were lysed to some extent by peritoneal BALB/c mouse PMN treated with this LK preparation. With Winn's assay, LK-treated PMN from BALB/c mice completely inhibited the growth of the admixed syngeneic tumor at a high effector to target ratio, when normal mice were used as recipients. When X-irradiated mice or nude mice were used as recipients, the tumor growth was partially inhibited by admixed LK-treated PMN, but the tumor began to grow gradually and finally killed the recipient mice, even when a high effector target ratio was used. When nude mice which had been given i.v. transfers of nylon wool column effluent spleen cells were used as recipients, the tumor inhibitory activity of LK-treated PMN was recovered to the same level as when normal mice were used as recipients. On the other hand, tumor inhibition by admixed LK-treated PMN in nude mice was not recovered by the transfer of X-irradiated nylon column effluent T-cells. As a mechanism of tumor inhibition by LK-treated PMN, a possible role of LK-treated PMN in reduction of tumor load is discussed.

Topics: 2-Chloroadenosine; Adenosine; Animals; Cells, Cultured; Female; Fibrosarcoma; Lymphokines; Male; Mast-Cell Sarcoma; Mice; Mice, Inbred BALB C; Mice, Inbred DBA; Mice, Nude; Neutrophils; Picibanil; Rats; Rats, Inbred Strains; T-Lymphocytes, Cytotoxic

Combined effects of cis-DDP and OK-432 on the ascites mastocytoma was studied in mice laying emphasis on the best timing for the administration of the OK-432. Mice were transplanted with FMA3 mastocytoma cells into the abdominal cavity at a dose of 10(5) cells per mouse. They were injected with cis-DDP next day at a single shot of 8 mg/kg in the abdominal cavity. A streptococcal preparation, OK-432, was i.p. injected two times at intervals of one week at a dose of 50 KE/kg per injection. The pair of injections started 1, 2, 3, 4 or 5 weeks after the transplantation. Mean survival time (M. S. T.) of mice was 16.8, 16.2 and 52.3 in the groups of mice nontreated, given OK-432 alone, and given cis-DDP alone, respectively. In comparison of M. S. T. within the groups of mice treated with the combination of cis-DDP and OK-432, the highest value was observed in the group which was given OK-432 between 3 and 4 weeks after the transplantation.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Biological Products; Cisplatin; Drug Administration Schedule; Male; Mast-Cell Sarcoma; Mice; Mice, Inbred Strains; Picibanil

Natural killer (NK) activity of mouse splenocytes was significantly augmented when the splenocytes were incubated for 3 to 4 hr with culture supernatants of mouse thymocytes stimulated by OK-432, an antitumor preparation from the Streptococcus pyogenes SU-strain. Antiviral activity was also detected in the culture supernatants, but IL 2 activity was not. When the culture supernatants of thymocytes stimulated by OK-432 were fractionated on a column of Blue Sepharose CL-6B, NK enhancing activity and antiviral activity were observed in partly overlapping fractions that bound to the column. However, the antiviral activity in the Blue Sepharose-bound fraction was neutralized completely by treatment with anti-IFN (alpha, beta) antiserum, whereas significant NK cell enhancing activity was still observed after treatment with anti-IFN (alpha, beta) antiserum. When the Blue Sepharose-bound fraction was subjected to gel filtration, the NK cell enhancing activity was detected in the 25,000 to 35,000 and 40,000 to 67,000 m.w. regions, but antiviral activity was observed in the over 67,000 m.w. region. These results indicate that a new kind of lymphokine, called natural killer cell activating factor (NKAF), distinct from IFN and IL 2, was found. The NKAF was found to have the following properties: its pI value is between pH 5.5 and 6.5, it binds to concanavalin A- and lentil agglutinin-Sepharose, and it is stable with pH 2-24 hr treatment. In addition, NKAF-producing cells were peanut agglutinin (PNA)-thymocytes when thymocytes were fractionated by the agglutination-sedimentation method with the use of PNA.

Topics: Animals; Biological Products; Cell Line; Culture Media; Cytotoxicity, Immunologic; Dose-Response Relationship, Immunologic; Female; G(M1) Ganglioside; Glycosphingolipids; Growth Inhibitors; Immune Sera; Interferons; Isoelectric Focusing; Killer Cells, Natural; Killer Factors, Yeast; Kinetics; Lectins; Lymphocyte Activation; Lymphoma; Mast-Cell Sarcoma; Mercaptoethanol; Mice; Mice, Inbred C3H; Molecular Weight; Peanut Agglutinin; Picibanil; Protein Biosynthesis; Proteins; T-Lymphocytes