Administration Of OK-432 Research Articles

Effect of OK-432 on large-bowel carcinogenesis in rats

To examine the carcinogenetic and growth inhibitory effects of OK-432, large-bowel carcinoma was induced experimentally in rats by intrarectal injection of N-methyl-N-nitrosourea (MNU), and OK-432 was administered intradermally. Rats were sacrificed after six months and the large intestine was cut into serial sections. Histopathologic examination and analysis of the infiltrating mononuclear cells, using monoclonal antibodies, were performed. The average rate of carcinogenesis per rat was 15.7 +/- 8.5 in the MNU alone group (n = 10) and 8.3 +/- 3.5 in the MNU and OK-432 group (n = 6). The invasion was deeper than the muscularis propria in 16 out of 157 lesions (10.2 percent) in the MNU alone group and in one out of 50 lesions in the MNU + OK-432 group (2.0 percent) (P less than 0.05). When time of appearance of atypical glands or carcinomas were compared in the MNU alone and MNU + OK-432 group, carcinogenesis was found to be delayed in the MNU + OK-432 group. In the investigation of infiltrating mononuclear cells using monoclonal antibodies, there were increases in helper T cells in both the MNU alone and MNU + OK-432 groups, but there was little difference between the two groups. The results of this study suggest that the suppression of experimental carcinogenesis in the large bowel by the concomitant administration of OK-432 with MNU, may be due to the enhanced activation or prolonged activated state of immunocompetent cells, which appear via antigen recognition, by OK-432.

Effect of combination therapy of radiation and local administration of OK-432 on a murine fibrosarcoma

The effects of a combination therapy of radiation and local administration of OK-432, which is one of BRMs (biological response modifiers), were studied using a radioresistant murine fibrosarcoma with weak immunogenicity. Fifty percent tumor control doses were 83.5 (79.6–87.4) Gy in animals given radiation alone and 64.3 (57.9–71.0) Gy in animals given OK-432 immediately after radiation, indicating that this biological response modifier can enhance the radiation dose effectiveness by a factor of 83.5 64.3 , that is, 1.30. Histological observations of treated tumors showed that the combination therapy induced a marked infiltration with lymphocytes and prominent degeneration and necrosis of the tumor cells. Examination of subsets of the infiltrating lymphocytes using the monoclonal antibodies showed that Lyt-2 positive lymphocytes were more abundant in the group given the combination therapy than in the radiation alone group on day 5 ( p > 0.05). Two days later, Lyt-1, Lyt-2, and L3T4 positive lymphocytes increased in the group given the combination therapy, whereas these lymphocytes disappeared in the group given radiation alone.

The endoscopic intratumor administration of OK-432 in gastric cancer unsuitable for surgery.

In eleven patients with gastric cancer who could not be operated on because of advanced age, refusal of surgery, or the presence of complications, the streptococcal preparation OK-432 was administered, both intradermally and by direct injection into the tumor under endoscopic guidance. Via gastric x-ray and endoscopic examinations, the tumor was observed to disappear in two cases-one advanced cancer and one early gastric cancer endoscopically. Particularly in the latter case repeated biopsies revealed no evidence of cancer cells. All patients developed fever after intratumor injection, but in no case was it necessary to discontinue administration, and no other serious side effects were noted. The endoscopic injection of OK-432 appears to be a safe and useful form of therapy for gastric cancer unsuitable for surgery.

Protection of OK‐432, a Streptococcus pyogenes Preparation, against Lethal Infection of Mice with Herpes Simplex Virus

We have studied the protective effect of OK-432, a biological response modifier (BRM) of Streptococcus pyogenes origin, on the lethal infection of mice with herpes simplex virus (HSV)-1. A single intraperitoneal (i.p.) injection of more than 10 micrograms of OK-432, when given at least two days before the infection, gave a marked effect yielding nearly 100% protection against ordinarily lethal infection. The protection was independent of the amount of infected virus inoculated. When given after the infection, the agent even at the maximal dose (100 micrograms), produced only a marginal effect. A single i.p. administration of OK-432 augmented the natural killer (NK) activity of peritoneal exudate cells and spleen mononuclear cells in mice 2 to 3 days after injection of OK-432, coinciding with the times when it induced a survival effect on HSV-infection. Treating OK-432-treated mice with a combination of an anti-macrophage agent, silica, and an anti-NK cell agent, anti-asialo GM1 serum, before infection diminished the antiviral effect of OK-432. The OK-432 protection against HSV infection was also markedly diminished in athymic nude mice. Thus, the protective effect of OK-432 on lethal HSV infection seems to be based on the activation of NK cells, macrophages, and T lymphocytes.

Inhibitory Effect of Toxoplasma Lysate Antigen on the Multiplication of Transplanted Tumor in Mice

Tachyzoites of the RH strain of Toxoplasma gondii were pretreated by a process of freezing and thawing followed by ultrasonication. After ultracentrifugation at 144,000 g for 120 minutes, the resulting supernatant contained a protein (TLA 144), which consisted mainly of a component of protozoan origin with a molecular weight of 10,000-20,000, and additional sugars, peptides, and amino acids. TLA 144 was soluble in water and of very low toxicity. Mice that had been inoculated with allogeneic (S-180) and syngeneic (Meth A) transplantable tumor cells, were injected intramuscularly with 100 micrograms of TLA 144 once a week for some time beginning one week after transplantation. It was found that after TLA treatment the multiplication of tumor cells was more intensely inhibited than following administration of OK-432, one of the biological response modifiers (BRMs).

STUDIES ON THE EFFECTS OF ORAL ADMINISTRATION OF OK-432 AS POSTOPERATIVE ADJUVANT IMMUNOTHERAPY

OK-432, a hemolytic streptococcal preparation, was given orally (11 cases) and intracutaneously (14 cases) to intramucosal gastric cancer patients as postoperative adjuvant immunotherapy. The following results were obtained. 1) Oral administration of OK-432 enhanced the skin reactions to PPD, Su-PS and Su-PR to an extent similar to that after intracutaneous administration of OK-432. 2) Oral administration of OK-432 caused greater increases in the absolute count and the OKT-3 positive cell ratio of peripheral lymphocytes than did intracutaneous administration of it. 3) Oral administration of OK-432 enhanced the PHA blastoid reaction of peripheral lymphocytes to a greater extent than did intracutaneous administration of it. 4) No side effects were noted after oral OK-432 administration. Thus, oral administration of OK-432 is considered to be a therapy which activates general cellmediated immune response, with effects that are comparable to those of intracutaneous administration of OK-432. Favorable results are anticipated upon further clinical evaluation of this extremely easy and safe dosing method.

Mechanism of Induction of Endogenous Tumor Necrosis Factor in Ascites of Ovarian Cancer Patients by OK-432, A Streptococcal Preparation

In four ovarian cancer patients with malignant ascites, 10 KE of OK-432 was intraperitoneally administered four times every other day for priming, and 40 KE of OK-432 in a single dose by the same route on day 13 for triggering. The changes in peripheral blood monocytes and intraperitoneal macrophages and the production of tumor necrosis factor (TNF) by peripheral blood mononuclear cells (PBMC) and ascitic lymphoid cells (ALC) were examined. In two of the four patients in whom TNF was induced in the ascites, the TNF production by PBMC and ALC was noted during priming, and after triggering, an increase in both the number of intraperitoneal macrophages and the TNF production by ALC was noted. In two other patients in whom TNF was not detected in the ascites, the ratio of intraperitoneal macrophages to ALC did not change throughout the whole period, and the TNF production by ALC was not augmented. These findings suggest that the priming administration of OK-432 can induce both intraperitoneal macrophages and peripheral blood monocytes into a primed state, and the triggering administration of OK-432 can increase the number of intraperitoneal OK-432-primed macrophages and induce TNF release from these cells.

The biological response modifier OK-432 (a streptococcal preparation) inhibits the development of autoimmune kidney disease in NZB/W F1 hybrid mice: possible involvement of tumor necrosis factor.

OK-432 (a streptococcal preparation) has been widely used for cancer immunotherapy in Japan. It is a potent immunostimulator, activating macrophages and T lymphocytes, and increasing the production of TNF and several other cytokines in both humans and animals. In the present study, we evaluated the prophylactic effect of OK-432 on the development of autoimmune kidney disease in NZB/W F1 (BWF1) mice. The mice were given 0.5 or 2.0 KE ('klinische Einheit'; clinical unit) of OK-432 intraperitoneally every week from 21 weeks of age to the time of death. The control group received the same volume of saline (vehicle). OK-432 delayed the development of proteinuria and prolonged the survival of these mice dose dependently. At 49 weeks, 33.3% of control mice were alive, whereas 55.6% in the 0.5-KE- and 75% in the 2.0-KE-treated mice were alive. In the control group, the serum cholesterol level increased due to the development of glomerulonephritis. In contrast, mice treated with OK-432 had significantly lower levels of serum cholesterol. The serum levels of anti-DNA and anti-TNP antibodies were not affected by OK-432 administration. OK-432 induced the production of tumor necrosis factor (TNF)-alpha in the peritoneal fluid in the BWF1 mice. These results indicate that the effect of OK-432 in preventing the development of autoimmune disease in the mice may result from the stimulation of the endogenous TNF-alpha production.

Usefulness of Preoperative Intratumor Administration of OK-432 in the Surgical Treatment of Gastric Cancer

Usefulness of Preoperative Intratumor Administration of OK-432 in the Surgical Treatment of Gastric Cancer

直腸癌に対する術前ＯＫ‐４３２の腫よう近傍投与による免疫学的研究 とくに主病巣および所属リンパ節の抗腫よう性反応について

直腸癌患者に対して, OK-432を術前腫瘍縁に局注し, 主病巣および所属リンパ節の抗腫瘍性反応の増強の可能性について免疫担当細胞の動態とNK細胞活性より検討した. 免疫担当細胞の動態は, 凍結切片の免疫組織化学染色および単離細胞のtwo color flow cytometric analysisにより検索した. NK活性は51Cr release assay法で測定した. OK-432の局注により腫瘍増殖先進部にLeu 4細胞が誘導され, そのsubsetsは主にLeu 3a+3b+細胞で, Leu 2a+細胞には変化を認めなかった. 所属リンパ節では, 遠位リンパ節にLeu 2a+細胞の相対的増加が認められ, とくにcytotoxic T cellおよびそのprecursorの増加がみられた. 腫瘍浸潤リンパ球のNK活性は, OK-432の局注により約半数の症例で増強した. 以上より, OK-432局注によって主病巣および所属リンパ節の細胞性免疫能が増強され, 癌治療での有用性が示唆された.

Antiherpes activity of the immunomodulator OK-432, a streptococcal preparation, in immunosuppressed mice

The antiviral activity of OK-432, an antitumor agent originating from Streptococcal preparations, against herpes simplex virus type 2 (HSV-2) was investigated in mice immunosuppressed by cyclophosphamide (CY). Intraperitoneal administration of OK-432 to mice 1 day after treatment with 200 mg CY/kg prevented death due to HSV-2 encephalitis in a dose-dependent manner. When the immunosuppressed mice were given OK-432 prior to HSV-2 infection, both by the intraperitoneal route, virus growth in the peritoneal cavity was significantly suppressed. Following with OK-432, the number of macrophages in immunosuppressed mice was increased to a significantly greater extent than the numbers of lymphocytes and polymorphonuclear leukocytes. The intrinsic antiviral activity of macrophages against HSV-2 as well as the natural killer (NK) activity against YAC-1 target cells was significantly enhanced by OK-432 in immunosuppressed mice.

The clinical efficacy of intratumoral OK-432 administration in advanced cancer patients

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.

Response to OK-432 (picibanil) in cancer patients and change of immunological parameters for the administration of OK-432 (picibanil)

We have been using streptococcal preparation OK-432 as a non-specific immunotherapeutic agent for head and neck cancers. OK-432 has been reported to elicit a generalized cellular humoral immune response. Recently many investigators have reported the potential usefulness of the Su-PS skin test reaction for evaluating the therapeutic response to OK-432. However, it is not clear how the Su-PS skin test actually reacts and what components of the Su-PS antigen, the Su-PS skin test reaction actually depends upon.In order to solve these questions, we investigated the relationship between the skin test reaction and leukocyte migration inhibition test using Su-Protein antigen and PPD antigen. Besides we measured the agglutinin titre against streptococcal cells as humoral immune response and examined the usefulness as a immunological parameter. Conclusions of this study are:1. The Su-PR skin test is a useful parameter since it generates essentially the same reaction as the Su-PS skin test does and also the Su-Protein antigen is much pure than the Su-PS antigen.2. The Su-PR skin test is a useful parameter for estimating the clinical response to the OK-432.3. The agglutinin titre against streptococcal cells is a reliable parameter for evaluating the response to the OK-432 injected transdermally.4. The agglutinin titre against streptococcal cells is not a reliable parameter for estimating the clinical effectiveness of the OK-432 therapy.

Significance of activation of reticuloendothelial function after hepatectomy in cirrhotic rats.

Effect of hepatectomy on the prognosis of cirrhotic rats prepared by oral administration of thioacetamide was studied from the standpoint of the reticuloendothelial function and energy metabolism of the liver. OK-432, a streptococcal preparation, was used to activate reticuloendothelial functions. Administration of OK-432 to cirrhotic rats prior to 70% hepatectomy significantly prevented the elevation of serum GOT, GPT and LDH, the prolongation of blood coagulation and the decrease of serum complement level. Hepatic ATP synthesis and RNA content were significantly increase by the use of OK-432. These findings suggest that activation of reticuloendothelial functions at the time of massive hepatectomy in cirrhotic rats may diminish hepatic injury, maintain serum complement level, and improve protein synthesis of the liver.

Antitumor effect of the streptococcal preparation OK-432 in a murine model of ovarian cancer.

The antitumor effects of the streptococcal preparation OK-432 were analyzed in a murine ovarian teratocarcinoma (MOT) model. Administration of OK-432 i.p. prevented tumor outgrowth in 75% of mice challenged with 10(3) MOT cells i.p. 24 h previously. Treatment was less successful in mice challenged with 10(4) or 10(5) cells, preventing tumor growth in 25% of the former and only 5% of the latter group. Tumor-challenged mice cured by injections of OK-432 were not rendered resistant to a subsequent challenge with 10(3) MOT cells 75 days after initial treatment. Only the i.p. route of administration was effective as i.v. OK-432 did not prolong survival of tumor-challenged mice. An antitumor response was detected as early as 24 h after i.p. treatment. This correlated temporally with an influx of neutrophils into the peritoneal cavity. Peritoneal cells obtained between 6 and 24 h after treatment were capable of lysing MOT targets in vitro. A single cell cytotoxicity assay demonstrated that peritoneal neutrophils, elicited by i.p. injection of OK-432, could bind to and lyse MOT targets. These data indicate that OK-432 is effective against small tumor cell inocula in this murine model of ovarian cancer and, furthermore, that the neutrophilic response into the peritoneal cavity plays a role in tumor rejection.

In vitro interferon producing activity of peripheral mononuclear cells in patients with chronic liver disease.

Interferon-gamma (IFN-gamma) was induced from a human peripheral mononuclear fraction by incubation with a streptococcal preparation stabilized with penicillin G (OK432). This IFN-gamma-producing activity was significantly reduced in patients with chronic hepatitis and hepatocellular carcinoma. In patients with liver cirrhosis it was also reduced but not significantly. Serum hepatitis B virus DNA and skin tests for the purified protein derivative of tuberculin, phytohemagglutinin-P and a polysaccharide fraction prepared from streptococcus pyogenes Su strain were determined to have no significant relation to this IFN-gamma-producing activity. Although the addition of interleukin 2 (IL-2) to the culture medium enhanced the IFN-gamma-producing activity, there was no difference in this enhancement between normal control and chronic hepatitis. Therefore reduction of the IFN-gamma-producing activity observed in chronic hepatitis seems to be caused by a decreased number of IFN-gamma-producing activity cells or hypofunction of these cells or both. Since HBeAg became negative in patients whose IFN-gamma-producing activity was increased by the administration of the immunopotentiator OK432 or IFN-beta, the IFN-producing system in the patients with B type hepatitis may contribute to the elimination of HBV. Adenine arabinoside suppressed IFN-gamma-producing activity both in vivo and in vitro.

Activation of reticuloendothelial function for prevention of endotoxemia after hepatectomy in cirrhotic patients.

In order to prevent endotoxemia after hepatectomy in cirrhotic patients, we administered OK-432 before and after hepatectomy to activate the reticuloendothelial function and studied its effect on postoperative endotoxemia. In the cirrhotic group without OK-432 administration (7 patients), the value of endotoxin increased significantly after hepatectomy, compared to the cirrhotic group which received OK-432 administration (5 patients) and the non-cirrhotic group (12 patients), and the endotoxin level was still higher than the preoperative value even on the 14th day. On the other hand, the cirrhotic group with OK-432 administration and the non-cirrhotic group showed minimal increases of endotoxin levels at the first day, which returned to the preoperative values at the third day. Base on these findings, it is suggested that activation of the reticuloendothelial function bears substantial significance as one of the therapeutic modalities for prevention of endotoxemia after hepatectomy in cirrhotic patients.

頭頚部悪性腫ように対するＯＫ‐４３２の長期生存効果について

In 82 patients with stage II or III malignant tumors of the head and neck, the cumulative survival rate was compared between 55 treated with and 27 not treated with OK-432. The annual survival rate was consistently higher in those treated than in those not treated with the agent; a significant difference was observed after 3 years and a slight different after 5 years.Intracutaneous administration of OK-432 had no effects on the cumulative survival rate, presumably because activation of pyrimidine fluoride compounds was unaffected.Of 57 parameters tested, we describe 9 immunological and biochemical parameters used in our hospital that indicate the onset and prognosis of malignant tumors of the head and neck. These parameters were:aspirin esterase, fibrinogen, a2-globulin, and cholinesterase levels; lymphocyte count, Su-Ps skin test, and PPD skin test; lymphocytes blastgenesis to PHA; and iron level.

The significance of activation of reticuloendothelial function on hepatectomy.

The prognosis of patients after massive hepatectomy is poor in certain cases whose hepatic reserve, including reticuloendothelial function, is deteriorated. We administered OK-432 before 70% hepatectomy on rats to activate the reticuloendothelial function and studied its effect on postoperative course. The elevations in plasma endotoxin, GOT and GTP were attenuated, and the deterioration of the complement activity after hepatectomy was greatly improved by OK-432 treatment. The RNA content in the liver was significantly increased by OK-432 administration. These findings suggest that activation of the reticuloendothelial function at the time of massive hepatectomy enhances endotoxin clearance from blood and thereby contributes in lessening the magnitude of hepatic injury, maintaining the serum complement, and improving liver protein synthesis.

PREOPERATIVE ENDOSCOPIC ADMINISTRATION OF OK-432 INTO LESIONS OF EARLY GASTRIC CANCER

OK-432, a killed preparation of Streptococcus pyogenes, was preoperatively administered under endoscope into lesions of early gastric cancer in 20 patients. Using these patients as subjects (treatment group). Local changes in the cancers, changes in cellular immunity of the hosts after administration, and the effect of OK-432 administration on anti-tumorous immune response in regional lymphnodes were investigated. Sixteen patients with early gastric cancer who did not received OK-432 were used as a control (non-treatment group). As a local histopathological finding, lymphoid cell infiltration in the tumors was often seen in the treatment group. Examination of cellular immunity before and after operation revealed that neither peripheral lymphocyte counts nor PHA-induced blastoid transformation of lymphocytes were changed in the treatment group, whereas these parameters were significantly reduced in the non-treatment group. Determination of anti-tumorous immune response in regional lymphnodes revealed that PHA-induced blastoid transformation and NK cell activity in proximal regional lymphnodes were significantly increased in the treatment group. When the microscopic structure of regional lymphnodes was examined, many patients in the treatment group showed high grades of SH and PH. From these results, the preoperative endoscopic administration of OK-432 into lesions of early gastric cancer seems to have immunological significance and clinical usefulness.