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PubMed1987

[Host mediated anti-tumor effect of Nocardia rubra cell wall skeleton on syngeneic tumor in mice].

The mode of action of Nocardia rubra cell wall skeleton (N-CWS) on Meth A fibrosarcoma (Meth A) was studied in BALB/c mice. N-CWS suppressed or regressed the intradermal growth of syngeneic Meth A cells in normal BALB/c and athymic BALB/c mice. The intradermally and subcutaneously infiltrated cells harvested from injection sites of N-CWS in normal mice showed in vitro cytotoxic activity against Meth A cells. Pretreatment of normal BALB/c mice with immunosuppressing agents such as hydrocortisone, carrageenan, or silica particles significantly reduced the anti-tumor effect of N-CWS. The growth of Meth A cells, rechallenged into BALB/c mice in which Meth A cells had once been suppressed or regressed by N-CWS treatment, was also inhibited, but not in similarly treated athymic nude mice. This resistant mechanism was shown to be dependent out cellular components but not on humoral components by the Winn Assay. The present results suggest that N-CWS exerts its anti-tumor activity by mediation of the immune system of the host and that the main effector cells in the early stage of tumor rejection are macrophages; T cells may also be involved in the later stage.

Animals↗
PubMed1987

Antitumour activity of purified arabinogalactan-peptidoglycan complex of the cell wall skeleton of Rhodococcus lentifragmentus.

Antitumour activity of arabinogalactan peptidoglycan (AP) complex (peptidoglycan and arabinogalactan liberated by an acid or alkaline treatment from Rhodococcus lentifragmentus AN-115 cell wall skeleton) was examined in mice and compared with that of the cell wall skeleton. The growth of syngeneic fibrosarcoma Meth A cells after implantation in BALB/c mice was significantly suppressed by AP complex, and also regressed after intratumoral injection of AP complex on days 1, 4 and 7 after tumour implantation. Although the activity of peptidoglycan was less than that of AP complex, peptidoglycan also showed both tumour-suppressive and regressive activities. Arabinogalactan did not show antitumour activity. It is interesting that peptidoglycan has an important role in the effect against tumours.

Animals↗
PubMed1976

Experimental immunotherapy of neoplastic pleural effusion with oil-attached BCG cell-wall skeleton in mice.

An experimental model for neoplastic pleural effusion was made using a transplantable pleural fibrosarcoma MC-106 in ddO mice, and a local immunotherapy of neoplastic pleural effusion with oil-attached BCG cell-wall skeleton was attempted. Viable cells (3 X 10(5)) of MC-106 were injected into the right pleural cavity of the mice on day 0 with a tuberculin syringe which was joined to a two-way tap attached to a capillary manometer. All of the mice in the control group, which received intrapleurally saline solution 24 hr after the injection of tumor cells, died within 24 days and the mean survival time was 16.9 +/- 3.4 (SD) days. Macroscopically massive blooded pleural effusion in both pleural cavities and multiple tumor nodules on the surface of parietal and visceral pleura were observed. On the other hand, the mice which received intrapleurally 100 mug of oil-attached BCG cell-wall skeleton 24 hr after the injection of tumor cells survived much longer. About 50% of the mice remained alive and were killed on day 95. They revealed histologically no malignant lesion of the pleura except for residual changes of inflammatory reactions.

Animals↗
PubMed1988

Effect of Nocardia rubra cell wall skeleton on interleukin 1 production from mouse peritoneal macrophages.

Nocardia rubra cell wall skeleton (N-CWS) was shown to augment interleukin 1 (IL-1) production from peritoneal resident and exudate macrophages in C3H/HeN mice. The mol. wt of the N-CWS-induced IL-1 product was about 17,000 daltons, which is a similar weight to that obtained by lipopolysaccharide stimulation. The stimulation of IL-1 production by N-CWS was seen as early as 8 h after the start of incubation and peak production was observed at 48 h. Profound effects were seen with 10 micrograms/ml or more of N-CWS. Experiments on the regulation of the N-CWS-augmented IL-1 production showed that prostaglandin E2 inhibited the augmentation, and indomethacin (cyclo-oxygenase inhibitor) further augmented it. Leukotriene B4 and AA861 (lipoxygenase inhibitor) had no effect. Our findings suggest that the previously reported adjuvant effect of N-CWS may, in part, be mediated via its ability to stimulate IL-1 production; and that such a stimulation may be blocked by prostaglandins.

Animals↗
PubMed1989

A possible mechanism of fever induced by Nocardia rubra cell wall skeleton (N-CWS) in experimental animals.

The characteristics of fever elicited by the cell wall skeleton of Nocardia rubra (N-CWS) and by lipopolysaccharide (LPS) were compared in rabbits, and the possible involvement of the antigenicity of N-CWS was investigated in guinea pigs. In rabbits, fever of more than 0.5 degree C developed after an intravenous (i.v.) injection of 10 micrograms/kg or more of N-CWS, and was monophasic with 30-100 micrograms/kg but biphasic with the highest dose of 300 micrograms/kg. LPS elicited fever with similar characteristics at doses of 0.01-0.1 microgram/kg. With both compounds, the fever was inhibited by indomethacin. Tolerance to N-CWS and LPS appeared after dosing with 30 or 0.1 micrograms/kg, respectively for 10 d. In guinea pigs sensitized with N-CWS, challenge with 1 or 10 micrograms/kg of N-CWS 10 d later, which did not induce fever in the nonsensitized animals, caused fever of more than 0.5 degree C, and delayed-type hypersensitivity (DTH) appeared. N-CWS also elicited fever in nonsensitized guinea pigs bearing N-CWS-sensitized lymphocytes or anti-N-CWS antibody; the fever was higher in the guinea pigs sensitized with the lymphocytes than in those with the anti-N-CWS antibody. In brief, single injections of N-CWS and of LPS elicited fever with similar characteristics, although the potency of N-CWS was weaker. With N-CWS, the fever is proposed to be triggered by the antigenicity of the compound itself, because doses as low as 1 or 10 micrograms/kg elicited fever along with immunological response in N-CWS-sensitized animals, but not in nonsensitized ones.

Animals↗
PubMed1986

Protective effect of Nocardia rubra cell wall skeleton on experimental infection in normal and immunosuppressed mice.

The protective effect of Nocardia rubra cell wall skeleton (N-CWS) on experimental infections was investigated in normal and immunosuppressed mice. Pretreatment with N-CWS provided protection against acute systemic infections due to Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Serratia marcescens and Listeria monocytogenes in normal mice. N-CWS administered before or after challenge also showed protective activity against Herpes simplex virus infection in normal mice. N-CWS was the most effective against extracellular parasitic Pseudomonas aeruginosa infection when administered 1 day before challenge, but displayed the most potent protective activity against infection with Listeria monocytogenes, a facultative intracellular parasite, when administered 7 to 14 days before challenge. In mice with subcutaneous Pseudomonas aeruginosa infection, N-CWS suppressed abscess formation and decreased the viable cell count in the resultant abscess foci when administered either intraperitoneally or subcutaneously to the infected site. In addition, treatment with N-CWS markedly restored host defense ability against pseudomonal infection in mice immunosuppressed with cyclophosphamide, hydrocortisone and X-ray irradiation.

Animals↗
PubMed1996

Lactobacillus- and bifidobacterium-mediated antigenotoxicity in the colon of rats.

Lactic acid bacteria (LAB) are proposed to have several beneficial effects, including the inactivation of carcinogens. We have studied the potential of Lactobacillus acidophilus (from a commercially available yogurt), Lactobacillus gasseri (P79), Lactobacillus confusus (DSM20196), Streptococcus thermophilus (NCIM 50083), Bifidobacterium breve and Bifidobacterium longum (from human infant stool) to prevent the induction of DNA damage by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG, 7.5 mg/kg body wt) in colon cells of the rat. Using the new technique of single cell microgel electrophoresis, all investigated strains were antigenotoxic toward MNNG after a single dose of 10(10) viable cells/kg body wt p.o. eight hours before the carcinogen. One-half and one-tenth of this initial dose resulted in a loss of protective activity. High doses of heat-treated L. acidophilus strains were also not antigenotoxic. One mechanism of the preventive effect could be that bacterial metabolites or components are responsible. Accordingly, selected examples were investigated in vitro in colon cells of the rat. Metabolically active L. acidophilus cells, as well as an acetone extract of the culture, prevented MNNG-induced DNA damage. Different cell fractions from L. acidophilus (cytoplasm, cell wall skeleton, cell wall) were devoid of antigenotoxic activity, whereas the peptidoglycan fraction and whole freeze-dried cells were antigenotoxic. As a second carcinogen, 1,2-dimethylhydrazine (DMH) was used. A dose- and time-response study was first performed to assess the effects of DMH in several segments of the gastrointestinal (GI) tract. Exposure for 16 hours to 15 or 25 mg DMH/kg body wt p.o. induced DNA damage in cells of the distal colon of rats, whereas no cytotoxicity was seen. Pretreatment orally with LAB on four consecutive mornings before DMH gavage (8 hours after the last LAB application) revealed that L. acidophilus, L. confusus, L. gasseri, B. longum, and B. breve inhibited the genotoxic effect of DMH. One of four S. thermophilus and one of three Lactobacillus delbrueckeii ssp. bulgaricus strains were also protective. Heat-treated L. acidophilus did not inhibit DMH-induced genotoxicity. A few aliquots of the colon cells were processed immunohistochemically for the presence of the "proliferation cell nuclear antigen" (PCNA). DMH treatment did not increase PCNA, nor was there any modulation by LAB. The effect of L. acidophilus on foreign compound-metabolizing enzymes (Phase I and Phase II) in liver and colon cells of rats revealed only one parameter to be modulated, namely, a two- to three-fold increase in the levels of NADPH-cytochrome P-450 reductase. The meaning of this finding, in terms of possible chemoprevention by LAB, remains unclear. In conclusion, our studies show that most, but not all, LAB tested could strongly inhibit genotoxicity in the GI tract of the rat and that viable LAB organisms are required for the protective effect in vivo. The comet assay technique is a powerful tool to elucidate such in vivo antigenotoxic activities in tumor target tissues.

1,2-Dimethylhydrazine↗
PubMed1984

Comparison of antimetastatic effect against Lewis lung carcinoma after intratumoral and intravenous injections of cell-wall skeleton of Propionibacterium acnes C7 in C57BL/6 mice.

Antimetastatic activity of cell-wall skeleton of Propionibacterium acnes C7 (P. acnes-CWS) in C57BL/6 mice varied depending on the injection route. The kinetics of antimetastatic effect against Lewis lung carcinoma (3LL) revealed that the sooner intratumoral (it) injection of P. acnes-CWS was carried out, the better the result. However, intravenous (iv) injection of P. acnes-CWS produced the best result if P. acnes-CWS was injected at about the time when metastases began to develop in the lungs. Pretreatment with intrafootpad (ifp) injection of P. acnes-CWS inhibited primary tumor growth and subsequent pulmonary metastases especially when given 7 days before tumor inoculation into the same footpad, but tended to enhance artificial pulmonary metastases when given 7 days before iv injection of tumor cells. In contrast, pretreatment with iv injection of P. acnes-CWS enhanced spontaneous pulmonary metastases especially when given 7 days before ifp inoculation of tumor cells, but inhibited artificial pulmonary metastases when given one day or 7 days before iv injection of tumor cells. These results suggest the difficulty of treatment of tumor metastases with immunological adjuvants. In T-cell-deprived mice, it injection of P. acnes-CWS showed no antimetastatic effect against 3LL, but iv injection was still effective. This indicates that T-cells are required for the antimetastatic effect of it injection of P. acnes-CWS, but iv injection of P. acnes-CWS is able to inhibit pulmonary metastases in T-cell-deprived mice.

Animals↗
PubMed1984

Inhibition of pulmonary metastasis by Nocardia rubra cell wall skeleton, with special reference to macrophage activation.

The antimetastatic activity of Nocardia rubra cell wall skeleton (N-CWS) with or without cyclophosphamide was examined in an experimental model of pulmonary metastasis induced by Lewis lung carcinoma in C57BL/6 mice. Lewis lung carcinoma cells were implanted into the footpads of mice, and the implanted tumors were removed 9 to 10 days later. Pulmonary metastatic nodules began to develop a few days after the implanted tumor was removed. The inhibitory effect of N-CWS was evaluated from the number of pulmonary surface nodules about 3 weeks after tumor implantation. The antimetastatic activity of N-CWS depended upon the dose, time, and route of its injection. Injection of N-CWS i.v. after removal of the implanted tumor caused the greatest inhibition of development of pulmonary metastases. Therapy with N-CWS plus cyclophosphamide prolonged significantly the survival of mice with metastases. The cytotoxic activities of peritoneal macrophages and macrophages in the lung against Lewis lung carcinoma cells were enhanced in mice treated with N-CWS. Injection i.v. of peritoneal macrophages activated with N-CWS inhibited pulmonary metastases. The role of macrophages in inhibition of micrometastasis in the lung is discussed.

Animals↗
PubMed1979

Activation of peritoneal macrophages by oil-attached cell-wall skeleton of BCG and Nocardia rubra.

The effect of oil-attached cell-wall skeletons of BCG (BCG-CWS) and Nocardia rubra (N. rubra-CWS) on macrophage functions was examined regarding phagocytosis of zymosan particles, uptake of 2-deoxy-D-glucose (2-dG), and cytostatic activity against syngeneic tumor cells. A single intraperitoneal injection of either BCG-CWS or N. rubra-CWS in mice resulted in accumulation of a large number of macrophages in the peritoneal cavity. They demonstrated, in vitro, a significant cytostatic effect on target cells as well as increase in phagocytic activity and uptake of 2-dG. These results suggest that one of underlying mechanisms for the antitumor activity of BCG-CWS and N. rubra-CWS is possibly the activated macrophage-mediated system.

Animals↗
PubMed1991

Antitumor effect of Nocardia rubra cell wall skeleton on syngeneically transplanted P388 tumors.

The antitumor activity of the immunomodulator, Nocardia rubra cell wall skeleton (N-CWS), was investigated using syngeneically transplanted P388 leukemia cells in a solid form. The s.c. growth of P388 tumors in DBA/2 mice was significantly suppressed by systemically administered N-CWS, and the effect was dose dependent. The antitumor effect of N-CWS was partially but significantly abrogated in splenectomized mice but not in T-cell or natural killer cell-deficient mice. Although spleen cells from mice treated with 1600 micrograms N-CWS contained no cytolytic activity, they exerted a significant cytostatic effect on P388 cell growth both in vitro and in vivo. Splenic cytostatic activity did not reside in T- or natural killer cells, but in plastic adherent cell population, macrophages. The response to N-CWS immunotherapy appeared to be associated with the number of macrophages infiltrating into the tumor lesions, and this was confirmed by histological analysis showing that P388 tumors from N-CWS-treated mice were intensively and dominantly infiltrated by macrophages. Furthermore, these were shown to be strongly tumor necrosis factor-positive by immunohistochemical analysis. These findings indicate that macrophages are the main effector cells playing a critical role in the suppression of P388 tumor growth in DBA/2 mice, and that tumor necrosis factor produced by these cells may be involved in the macrophage-mediated cytostatic effect induced by N-CWS. The fact that N-CWS suppressed the growth of weakly immunogenic P388 cells in syngeneic DBA/2 mice even when it was systemically injected would support the clinical potential of this agent.

Adjuvants, Immunologic↗
PubMed2001

Two receptor theory in innate immune activation: studies on the receptors for bacillus Culmet Guillen-cell wall skeleton.

Activation of the innate immune system is a prerequisite for the maturation of dendritic cells (DC) and macrophages (Mphi) followed by clonal expansion of the lymphocytes, targeting cells expressing "non-self' antigens. Microbes usually have a component competent to activate DC/Mphi for antigen presentation. This component has been called adjuvant, but recently renamed "pathogen-associated molecular pattern" (PAMP) or modulin based on its molecular identification. Here, we propose the hypothesis that DC/Mphi express two sorts of receptors for PAMP, whose signaling pathways lead to a sufficient antigen (Ag)-presenting state. In bacterial infection, a Toll-like receptor (TLR) and an uptake receptor participate in DC maturation and Mphi activation. Likewise, with a number of viruses, two of the receptors, with short consensus repeats (SCR), immunoglobulin-like domains or chemokine receptor-like motifs etc. induce functional modulation of DC/Mphi. In immune therapy for cancer, primary activation of the innate system would be essential for tumor Ag-specific T cell augmentation. Cancer cells express tumor-associated Ag but barely co-express PAMP, which situation does not allow for the activation of innate immune responses. Supplementing tumor-associated Ag with PAMP may be an effective therapy for patients with cancer. Here, we discuss the possibility of an innate immune therapy for cancer with reference to bacillus Culmet Guillen cell-wall skeleton (BCG-CWS).

Adjuvants, Immunologic↗
PubMed1983

[Immunotherapy of resectable lung cancer using Nocardia rubra cell wall skeleton].

A randomized controlled study of adjuvant immunotherapy was performed using Nocardia rubra cell wall skeleton (N-CWS) for inoperable lung cancer patients in 5 institutions from October, 1978 to June, 1981. All patients without pleural effusions were treated according to common protocol including chemotherapy and/or radiotherapy, and the patients on performance status 0 to 3 were randomized to control and N-CWS group. In N-CWS group, 400 micrograms N-CWS was injected once or twice into the bronchial tumor using fiberoptic bronchoscope, and sequentially 200 micrograms N-CWS was injected once a month into the skin from shoulder to upper arms. One year survival rate of 118 N-CWS-treated patients was excellent when compared to that of 108 control patients, but statistically not significant. According to histological type, survival period of the patients with small cell carcinoma was prolonged significantly in N-CWS group. The eligible patients with pleural effusions were initially randomized to control group and N-CWS group and treated under tube thoracostomy. In control group adriamycin was injected into the affected pleural cavity. In N-CWS group adriamycin and N-CWS were injected with the same method, and intracutaneous injections of 200 micrograms N-CWS were added once every month. Not only response rate for pleural effusions but also survival rate were statistically excellent in N-CWS group. Main adverse reactions of N-CWS were skin lesions in the injected sites and fever, but these were transient and not serious.

Adenocarcinoma↗
PubMed1983

[Effect of adjuvant immunotherapy with Nocardia rubra cell-wall skeleton in lung cancer].

A randomized clinical trial of intrapleural Nocardia rubra cell-wall skeleton (N-CWS) followed by intradermal N-CWS was performed against lung cancer patients from November, 1977 to June 1981. Totally, 190 patients were entered into this trial. The N-CWS treatment was effective in terms of prolongation of remission duration against not only operable but also inoperable patients. However, significant improvement of survival rate was observed only in operable patients, especially curative+relatively curative resection-group (p less than 0.05). The mode of recurrence was classified as local recurrence and distant metastasis in the curative+relatively curative resection-group. The rates of distant metastasis were 34.1 and 17.6%, respectively, in the control and the N-CWS groups. The rate of local recurrence was 13.6% in the control group, however, no local recurrence was observed in the C-NWS group. These results indicate the clinical effectiveness of the N-CWS treatment especially in curatively resectable lung cancer. No serious side effect was experienced during this trial.

Adenocarcinoma↗
PubMed1991

Synergistic effect of Nocardia rubra cell wall skeleton and recombinant interleukin 2 for in vivo induction of lymphokine-activated killer cells.

C57BL/6 mice inoculated i.p. with 3LL tumor cells were treated by local combination therapy with Nocardia rubra cell wall skeleton (N-CWS) and recombinant interleukin 2 (rIL-2). The combination treatment significantly prolonged their survival and augmented lymphokine-activated killer (LAK) activity of peritoneal cavity lymphocytes (PCL), compared with treatments with rIL-2 alone or N-CWS alone. After in vitro culture of peritoneal exudate mononuclear cells with rIL-2, the nonadherent population derived from N-CWS-injected tumor-bearing mice showed a significantly higher LAK activity than did that population derived from saline solution-injected mice. When N-CWS-induced PCL were cocultured with either N-CWS-induced macrophages or control macrophages in the presence of rIL-2, their LAK activity was higher than that of control PCL. Therefore, it was suggested that N-CWS-induced PCL themselves have a more potent ability as precursors of LAK cells. Phenotypic analysis on PCL populations revealed that N-CWS-induced PCL contained increased proportions of CD3+CD4-CD8- cells and asialo GM-1+ cells compared with control PCL. These results suggest that N-CWS selectively accumulates potent LAK precursors, namely, CD3+CD4-CD8- T-cells and asialo GM-1+ natural killer cells, at the injection site and that LAK cells are efficiently induced by subsequent administration of rIL-2.

Animals↗
PubMed1984

Randomized clinical trial of non-specific immunotherapy with cell-wall skeleton of Nocardia rubra.

A randomized clinical trial of intrapleural Nocardia rubra cell-wall skeleton (N-CWS) followed by intradermal N-CWS was performed against lung cancer patients from November, 1977 to June, 1981. Totally, 190 patients were entered into this trial. The N-CWS treatment was effective in terms of prolongation of remission duration against not only operable but also inoperable patients. However, significant improvement of survival rate was observed only in operable patients, especially curative operation group (p less than 0.05). The mode of recurrence was classified as local recurrence and distant metastasis in the curative operation group. The rates of distant metastasis were 34.1 and 17.6% respectively, in the control and the N-CWS groups. The rate of local recurrence was 13.6% in the control group, although no local recurrence was observed in the N-CWS group. These results indicate the clinical effectiveness of the N-CWS treatment especially in curatively resectable lung cancer. Major and frequent side-effects of the N-CWS treatment were fever up to 40 degrees C with an average duration of 2 days following intrapleural instillation and skin reaction at the site of intradermal injections. No serious side effect was experienced during this trial.

Adult↗
PubMed1991

Effect of cell wall skeleton and monophosphoryl lipid A adjuvant on the immunogenicity of a murine B16 melanoma vaccine.

We examined the effect of a new adjuvant consisting of purified mycobacterial cell wall skeleton (CWS) and monophosphoryl lipid A (MPL) on the immunogenicity of a murine melanoma vaccine. C57BL/6 mice were immunized to partially purified B16 melanoma vaccine given alone or together with different dose levels of adjuvant, or with saline or adjuvant alone. Humoral response, delayed-type hypersensitivity (DTH), in vitro cytotoxicity, and tumor-protective immunity to melanoma were measured following three biweekly immunizations. The adjuvant potentiated the antibody response to some, but not all, melanoma antigens in a dose-dependent fashion. The adjuvant also potentiated cellular immunity as measured by in vitro cytotoxicity assays. No potentiation of tumor-protective immunity was detected. In comparison to Freund's complete adjuvant, cell wall skeleton plus monophosphoryl lipid A (CWS:MPL) induced fewer cutaneous toxic effects and stronger antibody and DTH responses but resulted in no greater in vitro cytotoxicity or tumor-protective immunity. Thus, the adjuvant had a selective and dose-dependent effect on humoral responses to vaccine immunization but did not potentiate a tumor-protective immunity to B16 melanoma.

Adjuvants, Immunologic↗
PubMed1975

Immunotherapy of human malignant melanoma with oil-attached BCG cell-wall skeleton.

A first case of malignant melanoma treated with oil-attached BCG cell-wall skeleton (BCG-CWS) is reported. The patient, a 70-year-old farmer, with metastatic malignant melanoma was treated intralesionally with oil-attached BCG-CWS or was given intradermally with oil-attached BCG-CWS together with irradiated autologous and allogeneic tumor cells. Four months after the start of the immunotherapy with oil-attached BCG-CWS, Primary tumor and metastatic inguinal lymph nodes regressed markedly without any significant complications. Lymphocytosis in the peripheral blood was usually observed after injection of oil-attached BCG-CWS.

Adjuvants, Immunologic↗