Yasuyuki Takeda

Detection of serum cytokine levels in experimental cancer cachexia of colon 26 adenocarcinoma-bearing mice

The aim of this study was to evaluate the correlations between tumor size and cachexia parameters including cytokine levels in serum. In transplantable colon 26 adenocarcinoma-bearing mice, parameters having negative correlations with tumor size were host weight changes, epididymal adipose tissue weight, glucose and interleukin 3 (IL-3) concentration in serum. Parameters having a positive correlation with tumor size were the number of circulating white blood cells and immunosuppressive acidic protein (IAP), interleukin 6 (IL-6) and transforming growth factor beta (TGF-beta) concentration in serum.

Analysis of effector T cells against the murine syngeneic tumor MethA in mice orally administered antitumor polysaccharide SPR-901

The growth of MethA tumor was significantly inhibited by oral administration of the α-glucan SPR-901 in BALB/c (+/+) mice but not in nude mice. Mice treated orally with SPR-901 exhibited an augmentation of antigen-specific resistance against rechallenge with the tumor cells. The tumor-neutralizing activity of regional lymph node cells from MethA-bearing mice against the tumor was augmented by oral administration of SPR-901. The tumor-neutralizing activity of lymph node cells from SPR-901-treated mice mainly appeared in Lyt2+cells. Furthermore, lymphokine-activated killer activity of these cells was enhanced by administration of SPR-901. The antitumor effect of SPR-901 was abrogated in mice depleted of either L3T4+ or Lyt2+ cells, and in cyclosporin-A-treated mice. These results suggest that Lyt2+ cells are important effector cells in MethA-bearing mice orally adminstered SPR-901 and that functional exertion of both Lyt2+ and L3T4+T cells is necessary for the antitumor effect of orally administered SPR-901 in vivo.

Augmentation of host defense against Listeria monocytogenes infection by oral administration with polysaccharide RBS (RON)

Protection against Listeria monocytogenes in mice was enhanced by oral administration with a 30 mg/kg/day dose of polysaccharide RBS for 10 days. In mice treated with RBS, an enhanced elimination in vivo of L. monocytogenes was observed at the relatively late phase of listerial infection in correlation with enhanced immune responses against L. monocytogenes. The peritoneal macrophages from mice treated with RBS exhibited an increased activity of accessory cells for immune responses as assessed by production of interleukin-1 and antigen presentation to Listeria-immune T-cells. An increased activity of macrophages acting as accessory cells for immune responses may play important roles in the enhanced resistance against L. monocytogenes in mice treated orally with RBS.

Protective effect of SPR-901 (RBS) on the decrease of peripheral leukocyte number in 5-fluorouracil-treated mice

5-Fluorouracil (5-FU) induces a decrease in the number of peripheral leukocytes (leukopenia), which is one of the major obstacles in the chemotherapy of cancer. The number of peripheral leukocytes decreased by day 4 in mice injected i.p. with 130 mg/kg of 5-FU and recovered to the normal level by day 8. Such a decrease by 5-FU was prevented to some extent by the oral administration of 30 mg/kg/day of SPR-901. Proliferative responses of bone marrow cells to granulocyte/macrophage colony stimulating factor (GM-CSF) or granulocyte colony stimulating factor (G-CSF) were suppressed by 5-FU treatment and their recoveries were enhanced by SPR-901. The serum level of IL-6 in 5-FU-treated mice was increased by SPR-901. All of the mice treated with 300 mg/kg of 5-FU in combination with SPR-901 survived over 15 days, however, only 4 of 10 mice treated only with 300 mg/kg of 5-FU survived. These results suggest that SPR-901 acts on macrophages directly or indirectly, giving rise to the enhanced production of IL-1, IL-6, and other factors. Some of the factors derived from SPR-901 activated macrophages, perhaps mainly IL-6, act on the early stage of development of multipotent bone marrow progenitors synergistically with GM-CSF.

Effect of antitumor polysaccharide SPR-901 on antitumor activity in combination with 5-FU

We have examined the effects of the antitumor polysaccharide SPR-901 in combination with 5-fluorouracil (5-FU) on the antitumor activities against mouse syngeneic tumors. SPR-901 was administered p.o. from day 1 to day 10 at the dose rate of 30 mg/kg, and 5-FU was injected i.p. from day 1 to day 5 at the dose rate of 20 mg/kg after BALB/c mice were injected s.c. with 6 x 10(4) cells/mouse of Meth A on day 0. Tumor sizes of mice treated with both SPR-901 and 5-FU were significantly smaller than those from the untreated control group on days 10, 15 and 20. LAK activity of spleen cells from mice treated with both SPR-901 and 5-FU was higher than that from the untreated control group and either the SPR-901 or 5-FU treated group. Flow cytometrical analysis revealed that spleen cells from mice treated with both SPR-901 and 5-FU were much more abundant in both T-cell receptor alpha/beta+ and IL-2 receptor alpha+ T-cells. Furthermore, spleen cells from both the SPR-901- and 5-FU-treated groups exhibited higher growth responses to IL-2 than that from the untreated control group and either of the SPR-901- or 5-FU-treated groups. Therefore, the effects of the antitumor polysaccharide SPR-901 used in combination with 5-FU were augmented as compared with single drug use.

Augmentation of Host Defense Against Bacterial Infection Pretrfatjd Intraperitony with an α-Glucan RBS in Mice

AbstractProtection against Listeria mnocytogenes and Escherichia coli in mice was enhanced by an intraperitoneal (i.p.) administration of plysaccharide “RBS”. Eritoneal macrophages from mice adrmnistered i.p. with 30 mg/kg doses of RBS 4 days earlier exhibited increased scavenger functions as assessed by in vivo phagocytosis, in vitro intracellular killing and generation of superoxide anion. When cytokine production of the macrophages was assessed by biological assay and Northern blotting analysis, interleukin (IL)-l production and IG-1α gene expression were significantly increased in macrophages from RBS-treated mice. 01 the other hand, tumor necrosis factor (TNF)α gene was expressed in macrophages from RBS-treated mice at a much reduced level as compared with those in mice treated i.p. with Corynebacterium parvum on 4 days earlier. In correlation with expression of TNF gene in the mcrophages, RBS-treated mice were less susceptible to the lethal toxicity of LPS than C. parvum-treated mice. In RBS-treated...

Augmentation of Mitogen-Induced Interleukin 2 Production by Oral Administration of Polysaccharide SPR-901

The effects of oral administration of antitumor polysaccharide SPR-901 (RBS), an alpha-1,3 branched alpha-1,6 glucan, on concanavalin A (Con A)-induced interleukin 2 (IL-2) production of splenocytes were studied. The augmentation effect on Con A-induced IL-2 production was evident when more than 30 mg/kg of SPR-901 was administered orally to mice. On the other hand, oral administration of B512 dextran, an analogous alpha-1,6 glucan, did not show any augmentation effects on IL-2 production. The augmentation effect of SPR-901 on IL-2 production seemed to be mediated mainly by macrophages stimulated with SPR-901.

Effects of Stimulated or Immunologically Activated Macrophages on the Induction of Immune Responses to Listeria monocytogenes

The influences of peritoneal macrophages induced by proteose peptone, Corynebacterium parvum (C. parvum) or Bacillus Calmette Guérin (BCG) on the initiation and development of immune responses and protection against Listeria monocytogenes infection were studied in mice. Mice treated intraperitoneally (i.p.) with proteose peptone 4 days previously showed much the same level of protection against an intraperitoneal infection with Listeria as untreated mice. Mice treated i.p. with C. parvum 4 days previously, of which peritoneal macrophages had increased abilities for intracellular killing of Listeria and O2- generation as compared with peptone-elicited macrophages, exhibited an enhanced resistance against the listerial infection. The degree of immune responses, as assessed by delayed footpad reaction (DFR), was rather depressed in these mice because C. parvum-activated macrophages acting as scavenger cells reduced the amount of effective antigenic stimulation. BCG-activated peritoneal macrophages from mice treated i.p. with BCG 14 days previously showed a strong ability for antigen presentation in correlation with increases in the number of Ia-bearing macrophages and in the level of interleukin 1 (IL 1) production. These mice showed an early appearance of DFR response and a markedly enhanced resistance against the listerial infection. These results suggested that the differences in macrophage activities as scavenger cells, cytokine-secreting cells and antigen presenting cells may account for the differences in the responsiveness against listerial infection in peptone-, C. parvum- and BCG-treated mice.

Kinetics of fever and its related cytokines in mice after intraperitoneal infection with listeria monocytogenes

Abstract 1. 1. We studied the kinetics of fever and its related cytokine production in mice after an intraperitoneal (i.p.) infection with a sublethal dose of viable Listeria monocytogenes . By day 2 after an inoculation of Listeria , body temperature of the infected mice rose to a higher level than that of non-treated mice. On day 3 after infection, body temperature of the mice became lower than that of non-treated mice, and was accompanied by wasting. After day 5, the mice gradually recovered from this wasting to show a similar level of body temperature to the control mice. 2. 2. The number of peritoneal exudate cells (PEC) increased to the maximal level on day 3, when Thy1/CD3-positive cells emerged, and then the T cells increased to 35% of the non-adherent population of the PEC on day 8. By contrast, the number of IgM + - or Ly1 + -positive cells was at a constant level (∼60%) and ∼20%, respectively) of the non-adherent PEC during the infection. 3. 3. To characterize the productive pattern of fever-related cytokines in listeriosis, we investigated their expression in the PEC from mice during infection. In the adherent-PEC, IL-1α and IL-6 mRNAs were consistently expressed from day 1 to day 8 after infection, while TNF had already been expressed at the highest level on day 1 and decreased on day 8. IFN-γ mRNA was first detected in the non-adherent PEC on day 3 and increased on day 8. 4. 4. These results suggest that TNF may be related to the initial febrile state and the consecutive wasting at the early phase of listeriosis, followed by the T-cell expansion producing IFN-γ for elimination of Listeria .

Biliary secretion of antibody to dextran following oral immunization with dextran B512

Anti-dextran in bile was induced to high levels by oral immunization with dextran B512. IgM anti-dextran were dominant in serum, whereas IgG anti-dextran was dominant in bile. The binding properties of these IgM and IgG antibodies were different, as determined by ELISA with several dextrans. Splenocytes produced equal amounts of IgG and IgM antidextran but cells from mesenteric lymph nodes (MLN) and Peyers patches produced mainly IgG anti-dextran. Differences were observed among different strains of mice in their ability to produce anti-dextran in serum and bile upon immunization with dextran. BALB/c mice, which are intermediate responders in terms of their serum antibody levels, produced high levels of anti-dextran in bile. C3H/He and C57BL/6, which are high responders in terms of serum antibody levels, had intermediate responses in bile. DBA/2, which are low responders in terms of serum antibody levels, showed low responses in bile. The results provide further evidence of the existence of anti-dextran producing cells. These results indicate that B cells in systemic and mucosal-associated lymphoid tissues from BALB/c, C3H/He, C57BL/6 and DBA/2 mice respond differently to oral immunization with dextran B512.