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PubMed1979

Comparative study on antitumor effect of cell-wall skeleton of Mycobacterium bovis BCG and Nocardia rubra, with reference to T-cell dependency and independency.

Antitumor activity induced by the oil-attached cell-wall skeleton of Nocardia rubra (N-CWS) was compared with that of the oil-attached cell-wall skeleton of Mycobacterium bovis BCG (BCG-CWS) in syngeneic BALB/c tumor-host systems. In normal BALB/c mice (+/+), N-CWS exhibited stronger suppressive effect on syngeneic Br-1 and MCA tumors than did BCG-CWS. In athymic nude mice (nu/nu), BCG-CWS was as effective as N-CWS for the suppression of growth of such tumors. Suppressive effect of N-CWS treatment appears to be stronger to some extent in +/+ mice than in nu/nu mice. Immune spleen cells obtained from +/+ mice after footpad inoculation of MCA tumor cells mixed with N-CWS were effective in suppressing the MCA tumor growth, although those obtained from mice after inoculation of MCA tumor cells mixed with BCG-CWS did not exhibit a suppressive effect. This antitumor activity of immune spleen cells may be attributed to tumor-specific killer T cells. The differences of antitumor activities induced by these agents were discussed with reference to T-cell dependency and independency.

Animals↗
PubMed1989

Effects of periodic administration of Nocardia rubra cell-wall skeleton on immunoglobulin production and B-cell-stimulatory factor activity in vitro in workers at a poison gas factory.

The former workers at the Okunojima poison gas factory (poison gas workers) are a high-risk group for malignant neoplasms and show abnormalities in cellular immunity. At the same time, poison gas workers often have chronic respiratory diseases, such as chronic bronchitis, and are highly susceptible to respiratory infections. To explore the possibility of immunological cancer prevention, we have periodically administered 200 micrograms Nocardia rubra cell-wall skeleton (N-CWS) to poison gas workers once every 3 months since December 1978. During this period, we noted a significantly lower incidence of influenza among poison gas workers receiving N-CWS than in those not receiving the drug during the influenza epidemic. This finding suggested that the administration of N-CWS enhanced the resistance of these workers to infections. Therefore, periodical administration of N-CWS to poison gas workers was considered to enhance the reduced T-cell function of normalizing antibody production by stimulating the production of B-cell-stimulatory factor (BSF). In the present study, to clarify the mechanism of immunosuppression in the poison gas workers and to examine the effects of continual administration of N-CWS on this condition, we compared the immunoglobulin production and the proliferative and differentiative activities of B-cell-stimulatory factor (BSF) of peripheral blood mononuclear cells (PBMC), in poison gas workers treated or not treated with N-CWS. Comparisons were also made with age-matched healthy controls. In the untreated poison gas workers, immunoglobulin and BSF production of PBMC were reduced as compared with the control group. On the other hand, in the poison gas workers receiving N-CWS, immunoglobulin and BSF production of PBMC were restored nearly to the control level. These results show that in vitro antibody production in the poison gas workers was reduced and that a reduction in BSF production of T cells was one of its causes.

Adjuvants, Immunologic↗
PubMed1979

Partial inhibition of the growth of Mycobacterium lepraemurium in C3H mice immunized with cell wall skeletons.

C3H mice stimulated with M. lepraemurium cell wall skeletons (LM-CWS) were challenged with viable M. lepraemurium, and delayed-type hypersensitivity (DTH) to picryl chloride was measured. In one type of experiment the mice were challenged at the time when stimulation of cell-mediated resistance by means of LM-CWS was undertaken. The major purpose was to investigate principles pertaining to immunotherapy. In contrast to the loss of a detectable DTH response to picryl chloride, development of murine leprosy was partially suppressed. In the second type of experiment the mice were stimulated with oil-attached LM-CWS seven weeks before challenge with M. lepraemurium. Findings were: a) that nonspecific DTH as measured by sensitization and challenge with picryl chloride was activated before the infection with M. lepraemurium and b) that the DTH which developed was associated with partial protection against the growth of lepromata. The murine leprosy which developed in C3H mice stimulated with LM-CWS was progressive after some delay.

Animals↗
PubMedDate not supplied

Phase I study with cell wall skeleton of Nocardia rubra.

Toxicity of the cell wall skeleton of Nocardia rubra (N-CWS) was evaluated in 60 patients with lung cancer, leukemia, and malignant melanoma who were given the agent intratumorally, intrapleurally, and intradermally in doses of 100-2000 micrograms. Fever was the most common toxic effect and was more frequent after intratumoral and intrapleural doses than after intradermal doses. The other predominant reaction was skin lesions such as erythema, induration, and abscess at the injection sites.

Abscess↗
PubMed1976

Effect of oil-attached BCG cell-wall skeleton on the induction of pleural fibrosarcomas in mice.

The effect of oil-attached BCG cell-wall skeleton on the induction of pleural fibrosarcomas by 3-methylcholanthrene in mice was examined. The pleural fibrosarcomas were induced by a substernal injection of 3-methylcholanthrene into the thoracic cavity of ddO mice. One week after the injection of 3-methylcholanthrene, 100 mug of oil-attached BCG cell-wall skeleton was injected subcutaneously every week for 10 weeks. In the observation period of 130 days, the incidence of pleural fibrosarcoma was 67% in the control mice and 39% in the mice treated with BCG cell-wall skeleton. The latent period of tumor induction was prolonged in the mice treated with BCG cell-wall skeleton.

Animals↗
PubMed2004

Gene-inducing program of human dendritic cells in response to BCG cell-wall skeleton (CWS), which reflects adjuvancy required for tumor immunotherapy.

Adjuvants induce the expression of a number of genes in dendritic cells (DCs), which facilitate effective antigen-presentation and cytokine/chemokine liberation. It has been accepted that the toll-like receptor (TLR) family governs the adjuvant activity in DCs. An adjuvant with a long history is mycobacteria in an oil-in-water emulsion, namely Freund's complete adjuvant. Since the active center for the adjuvancy in mycobacteria is the cell-wall skeleton (CWS), we used the bacillus Calmette-Guerin cell-wall skeleton (BCG-CWS) to test DC maturation by GeneChip analysis. We identified the genes supporting an efficient DC response and output. Approximately 2000 genes were up-regulated by BCG-CWS stimulation. BCG-CWS-, peptidoglycan (PGN)- and lipopolysaccharide (LPS)-stimulation generally up-regulated some gene clusters including genes for inflammatory cytokines (TNF, IL1alpha, IL1beta, IL6, IL12 p40, IL23 p19, etc.), chemokines (CCL20, IL8, etc.), cell adhesion molecules (ICAM-1, etc.), apoptosis-related proteins (GADD45B, BCL2A1, etc.), metabolic enzymes (PTGS2, SOD2, etc.) and miscellaneous proteins (EHD1, TNFAIP6, etc.). LPS-stimulation, but not BCG-CWS- or PGN-stimulation, up-regulated the interferon-inducible antiviral proteins, including IFIT1, IFIT2, IFIT4, CXCL10, ISG15, OASL, IFITM1 and MX1. We also found that the BCG-CWS- or PGN-stimulation up-regulated CXCL5, MMP1, etc. We discussed their properties in association with TLRs and recently discovered TLR adapters.

Adjuvants, Immunologic↗
PubMed1989

Augmentative effect of Nocardia rubra cell-wall skeleton on the induction of human lymphokine-activated killer (LAK) cells by the production of LAK cell helper factor(s).

Nocardia rubra cell wall skeleton (N-CWS) was found to synergistically augment lymphokine-activated killer (LAK) cell generation from human peripheral blood mononuclear cells (PBMC) in the presence of a suboptimal dose of recombinant interleukin-2 (rIL-2). N-CWS increased the number of PBMC expressing IL-2 receptor on their surfaces, and the presence of N-CWS at the early stage of the culture period was essential for the exertion of its augmentative activity on the LAK induction. The predominant phenotype of LAK precursor cells responding to N-CWS and rIL-2 was CD3- CD16+. Culture supernatant from N-CWS-stimulated PBMC was found to act as a substitute for N-CWS in the induction of LAK generation in the presence of rIL-2, suggesting that these cells produced a factor capable of augmenting LAK cell induction (LAK helper factor, LHF). LHF was found to have a molecular mass of 29 kDa by gel filtration, and could also function as a killer helper factor to augment allo-antigen-specific cytotoxic T lymphocyte generation from human peripheral blood T cells as well as murine thymocytes. LHF showed no species specificity, indicating that it is different from IL-4. The enhancing activity of LHF was not neutralized with anti-TNF alpha, anti-IL-1 alpha, or anti-IL-1 beta antibodies. Furthermore, no tumor necrosis factor-alpha (TNF alpha), TNF beta, IL-1 alpha, beta, IL-2, IL-5, IL-6 or interferon activity was detected in semi-purified LHF during enzyme-linked immunosorbant assay and biological assays. The present findings indicate that LHF produced from N-CWS-stimulated PBMC is a molecule distinct from TNF alpha, TNF beta, interferon, IL-1, -2, -4, -5, and -6, and suggest that LHF might be a novel lymphokine involved in LAK generation.

Adjuvants, Immunologic↗
PubMed1986

[Immunotherapy of acute myelogenous leukemia (AML) in adults with the Nocardia rubra cell wall skeleton].

The effect of immunotherapy with Nocardia rubra cell-wall skeleton (N-CWS) on the remission duration and survival of adults with acute myelogenous leukemia (AML) was studied in a prospective randomized controlled study. After having been induced into complete remission and consolidated, 73 patients were randomized either to maintenance chemotherapy or maintenance chemotherapy plus immunotherapy with N-CWS and irradiated allogeneic AML cells. Thirty-four patients in the chemotherapy group and 32 in the chemoimmunotherapy group were evaluable. Six months after the closure of the study, the immunotherapy showed a borderline beneficial effect on remission duration (p = 0.080) and on survival length (p = 0.098). When the data were analyzed at 42 months after entry, there was a borderline significant difference in remission duration (p = 0.066) between the two groups, prolonging the 50% remission period by 120 days, but no significant differences in survival length (p = 0.306), although the 50% survival was 168 days longer in the chemoimmunotherapy group. Thus, immunotherapy with N-CWS and irradiated allogeneic AML cells seems to be active in the treatment of adult AML when used for maintenance therapy in combination with chemotherapy.

Adult↗
PubMed1981

Killing of tumor cells in vitro by macrophages from mice given injections of squalene-treated cell wall skeleton of Nocardia rubra.

Peritoneal exudate cells (PEC) harvested from mice after i.p. injection of squalene-treated cell wall skeleton of Nocardia rubra (N. rubra-CWS) demonstrated vigorous cytolytic activity in vitro toward tumor target cells. Fractionation of these PEC by adherence to plastic dishes showed that the cytolytic activity in PEC was associated with an adherent phagocytic cell. Induction of the cytolytic adherent PEC required an optimal dose of 50 micrograms N. rubra-CWS and i.p. injection. Cytolytic activity of N. rubra-CWS-induced adherent PEC was maximal after 5 days and fell steadily thereafter. Susceptible tumor targets included cells syngeneic, allogeneic, and xenogeneic to the effector cell source. In contrast, nonneoplastic xenogeneic cells were not affected by N. rubra-CWS-induced adherent PEC. The effector cells were not found in the spleen or peripheral lymph nodes. In addition, the cytolytic activity of N. rubra-CWS-induced adherent PEC was completely inhibited by treatment with antimacrophage serum and complement or carrageenan. Treatment with monoclonal anti-Thy 1.2 antibody and complement, however, did not affect the cytolytic activity of the adherent PEC. These features make it likely that N. rubra-CWS-induced cytolytic effector cells are macrophages.

Animals↗
PubMed1975

Adjuvant activity of mycobacterial fractions. I. Purification and in vivo adjuvant activity of cell wall skeletons of Mycobacterium bovis BCG, Nocardia asteroides 131 and Corynebacterium diphtheriae PW8.

The adjuvant activity of cell wall skeletons (mycolic acid-arabino-galactan-mucopeptide, CWS) prepared from the cells of mycobacteria, nocardia and corynebacteria was examined in vivo in mice and guinea pigs. The cell wall skeletons of Mycobacterium bovis BCG (BCG-CWS), Nocardia asteroides 131 and Corynebacterium diphtheriae PWC suspended in Freund's incomplete adjuvant (FIA) as water-in-oil emulsions showed potent adjuvant activity on the formation of circulating antibody and cell-mediated immunity to bovine serum albumin (BSA), sheep erythrocytes (SRBC) and sulfanylazo-bovine serum albumin (SA-BSA) in mice and guinea pigs. After acetylation or acid treatment, BCG-CWS retained its adjuvant activity, but the activity of BCG-CWS was destroyed completely by alkaline treatment. The cell wall constituents, arabinose-mycolate and arabino-galactan, prepared from BCG-CWS showed no adjuvant activity. It was also shown that BCG-CWS suspended in phosphate buffered saline or associated with oil droplets augmented cell-mediated cytotoxicity in which thymus-derived lymphocytes (T-cells) are effector cells.

Acids↗
PubMed1978

Antitumor activity of peritoneal exudate cells induced by cell-wall skeleton of Mycobacterium bovis BCG.

Peritoneal exudate cells (PEC) induced by oil-attached cell-wall skeleton of Mycobacterium bovis BCG (BCG-CWS) in ACI/N rats were tested for their effect on both in vivo and in vitro growth of syngeneic fibrosarcoma cells (AMC-60). Treatment of rats with intraperitoneal injections of BCG-CWS induced regression of syngeneic ascites tumor and increased the number of survivals. Whole PEC and adherent PEC from rats injected intraperitoneally with BCG-CWS inhibited the uptake of tritiated thymidine into the fibrosarcoma cells in an in vitro cytostasis test. This in vitro cytostatic effect was more marked as the ratio of effector to target cells increased. In addition, when tumor cells were inoculated subcutaneously with BCG-CWS activated PEC, tumor takes decreased markedly. Oil-stimulated PEC and normal peritoneal resident cells were inactive in inhibition of tumor growth in vivo and in vitro.

Animals↗
PubMed1986

Augmented production of colony-stimulating factor in C3H/HeN mice immunized with Nocardia rubra cell wall skeleton.

C3H/HeN mice subcutaneously injected repeatedly with Nocardia rubra cell wall skeleton (N-CWS) acquired cellular immunity against N-CWS. In these N-CWS-immunized mice, the serum colony-stimulating factor (CSF) and granulocyte-macrophage colony-forming units in the bone marrow remarkably increased after an intraperitoneal (i.p.) injection of N-CWS compared with those in normal control mice given an single i.p. injection. To analyze the effects of secondary immune response to N-CWS on CSF production by splenocytes, whole mononuclear leukocytes (WMNL), nylon fiber-nonadherent splenocytes (NADC), and plastic-adherent splenocytes (ADC) were cultured in the presence of N-CWS, and then the supernatants of each cell fraction were assayed for CSF. A fraction of WMNL from immunized mice was found to produce more CSF than WMNL from control mice, and either fraction of NADC or ADC separated from WMNL produced markedly less potent CSF when cultured separately. However, when NADC from immunized mice were cultured with ADC from either immunized or normal mice, CSF production recovered to the level shown by WMNL. The role of ADC could be substituted for by cell-free culture medium of either ADC plus N-CWS or the J774 cell line, in which high interleukin-1 activity was detected. A surface marker study showed that depletion of either Lyt-1.1+ or Lyt-2.1+ cells caused a striking loss of CSF production. These data suggested that CSF in the sera of immunized mice challenged with an N-CWS i.p. injection is mainly produced by N-CWS-sensitized T lymphocytes with help of a macrophage-derived humoral factor(s).

Adjuvants, Immunologic↗
PubMed1981

Effect of Bacillus Calmette-Guérin cell wall skeleton on the induction of the cytotoxic and suppressor T cells against syngeneic tumor in the mouse.

The effect of Bacillus Calmette-Guérin cell wall skeleton (BCG-CWS), which is one of the well-established immunopotentiators, on the induction of either cytotoxic or suppressor T cells against syngeneic tumor S1509a was investigated by an in vitro assay system. BCG-CWS showed an apparent augmenting activity in the induction of cytotoxic T cells against a syngeneic tumor when it was administered subcutaneously into mice with mitomycin C (MMC)-treated tumor cells in a form of oil-in-water emulsion. In addition, cytotoxic T cell activity was dominantly generated in regional lymph nodes. Furthermore, the immunized animals developed stronger resistance against the second inoculum of the same viable tumor cells. However, BCG-CWS had no effect on the induction of suppressor T cells in tumor-bearing host (TBH), and could not induce cytotoxic T cells against the homologous tumor in TBH in which suppressor T cells had been rapidly activated. The growth of tumor was neither suppressor nor enhanced, even if an oil-attached BCG-CWS was administered in TBH.

Animals↗
PubMed1984

Potentiation of tumoricidal properties of murine macrophages by Nocardia rubra cell wall skeleton (N-CWS).

Intraperitoneal injection of squalene-treated cell wall skeleton of Nocardia rubra (N-CWS) caused increase in number of peritoneal exudate cells (PEC). Adherent macrophages obtained from N-CWS-treated PEC suppressed growth of methylcholanthrene-induced fibrosarcoma (Meth-A), when injected intradermally with the tumor cells into BALB/c mice. The macrophages showed strong cytotoxicity against Meth-A cells in vitro. When treated with 10 micrograms/ml of N-CWS in vitro, proteose peptone-induced macrophages acquired tumoricidal property but resident macrophages showed no cytotoxicity after the treatment. In the supernatant of spleen cells cultured for 72 hours in the presence of N-CWS (10 micrograms/ml), the presence of (a) factor(s) with macrophage activating effect was observed. This factor, shown to be identical to macrophage activating factor (MAF) in molecular weight, showed synergy with N-CWS in potentiating macrophage cytotoxicity against tumor cells.

Adjuvants, Immunologic↗
PubMed1982

The restoration of suppressed T cell activities induced by 7,12-dimethylbenz[a]anthracene with an anti-tumor immunomodulator, BCG-cell wall skeleton.

The restoring effect of Bacille de Calmette et Guérin (BCG)-cell wall skeleton (CWS) on the suppressed T cell reactivities in mice treated with 7,12-dimethylbenz[a]anthracene (DMBA) was studied using a system involving the induction of hapten-reactive cytotoxic T cells and mixed lymphocyte reaction in vitro. When the mice were treated with DMBA, the reactivity of lymphoid cells to generate cytotoxic T cells and proliferative T cell responses against allogeneic spleen cells was markedly suppressed. When BCG-CWS was administered to mice by intravenous injection before and after the treatment with DMBA, these depressed T cell functions reverted completely to the normal level. However, the restoration of the immune functions by BCG-CWS varied depending on the strain of mice. Namely, the reactivity of C57BL/6 mice was restored by BCG-CWS, whereas that of C3H/He mice was not. This strain difference correlates very well with the strain difference in effectiveness of the prevention of the DMBA-induced tumorigenesis by BCG-CWS. The possible relation between the restoration of T cell reactivities by BCG-CWS and prevention of tumorigenesis by BCG-CWS is discussed in the context of the role of host immune surveillance in the regulation of tumor development.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1989

Synergy of Nocardia rubra cell wall skeleton and interleukin 2 in the in vivo induction of murine lymphokine-activated killer cell activity.

Combination of an ip injection of Nocardia rubra cell wall skeleton (N-CWS) and 3 daily sc injections of human recombinant interleukin 2 (rIL 2) into C3H/HeN mice resulted not only in a significant increase in the number of peritoneal cells (PC) but also in a potent induction of their lymphokine-activated killer (LAK) activity, compared with results obtained with N-CWS or rIL 2 alone. The augmented LAK activity of PC was mediated by nonadherent, nonphagocytic, Thy-1.2+(-)- and asialo GM1+ cells. Nonadherent PC induced by an ip injection of N-CWS bound more 125I-labeled rIL 2 than did normal, nonadherent PC, and generated high LAK activity when cultured overnight with rIL 2. In contrast, normal, nonadherent PC responded only weakly to the overnight stimulation with rIL 2. The phenotype of N-CWS-induced PC with an elevated IL 2 responsiveness was Thy-1.2+(-)-, Lyt-1.1-, Lyt-2.1- and asialo GM1+, suggesting that the N-CWS-stimulated LAK precursors were derived mainly from the NK cell lineage. However, mature T cells may also be involved in this mechanism, because N-CWS failed to augment the IL 2 responsiveness of nonadherent PC in BALB/c nu/nu mice. Treatment of C57BL/6N mice bearing solid Lewis lung carcinoma (3LL) tumors with an intratumoral injection of N-CWS followed by 6 daily sc injections of rIL 2 resulted in the apparent suppression of tumor growth, while N-CWS or rIL 2 alone produced no such suppression. These results suggest that N-CWS augments the antitumor effect of rIL 2 by accumulating LAK precursors and elevating their responsiveness to rIL 2 at the injection site.

Adjuvants, Immunologic↗
PubMed1979

Properties of interferon induced by purified protein derivative of tuberculin in mice sensitized with BCG or cell-wall skeleton of BCG.

Mice sensitized with either BCG or cell-wall skeleton of BCG (BCG-CWS) produced interferon in blood after stimulation with specific antigen, purified protein derivative of tuberculin (PPD). Both BCG-infected normal (C57BL/6) and thymic nude (BALB/c, nu/nu) mice showed enhanced activity to produce interferon by stimulation with E. coli endotoxin. However, detectable interferon was not produced in athymic nude mice sensitized with BCG or BCG-CWS by stimulation with PPD. Immune-induced interferon (I-IF) produced by BCG-CWS and PPD in mice was different in biological and physicochemical properties from virus-induced interferon. I-IF showed about 100 times more potent L-cell growth inhibitory activity than virus-induced L-cell interferon (L-IF). Both I-IF and L-IF showed macrophage-activating activity, which renders resting macrphages cytotoxic to L1210 leukemia cells. Antiviral and macrophage-activating activity of interferon preparation was not separated physicochemically in this study.

Animals↗