Administration Of OK-432 Research Articles

Enhancement of immune reactivities of regional lymphnodes by oral administration of OK-432 in colon cancer patients

Effect of oral administration of OK-432 on the immune reactivities of regional lymphnode (RNLs) and peripheral blood lymphocytes (PBLs) in colon cancer patients were investigated. Twenty KE of OK-432 was given orally on 7 days prior to the operation, and peripheral blood and regional lymphnodes were collected at the time of operation. As for the immune reactivities of PBLs such as lymphocyte proliferative response to phytohemagglutinin (PHA), natural killer (NK) cell activity and Concanavalin A (Con-A) induced suppressor cell activity, there were no significant differences between patients with oral OK-432 (OK-432 (+) group) and those without OK-432 (OK-432 (-) group). NK cell activity of RLNs was significantly higher in OK-432 (+) group than in OK-432 (-) group. Furthermore, the decline of Con-A induced suppressor cell acivity was found in OK-432 (+) group. T cell subsets in regional lymphnode tissues were identified with the use of monoclonal antibodies and immuneperoxidase technique. Both of Leu-2 a and Leu-7 positive cells distributed more frequently in lymphnodes from OK-432 (+) group than in those from OK-432 (-) group. Thus, these results suggested that immune reactivities of regional lymphnodes were en-hanced by oral administration of OK-432 in colon cancer patients.

THE USEFULNESS OF THE Su-PS AND Su-PR SKIN REACTION TEST FOR GASTRIC CANCER PATIENTS

The Su-PS and Su-PR skin reaction was determined in 144 patients with gastric cancer, and the following results were obtained. 1) The Su-PS and Su-PR skin reaction was found to be a possible parameter for determination of immunological competence accompanying the advance of gastric cancer. 2) The Su-PS skin reaction was a parameter representing the sensitivity of gastric cancer patients to OK-432. 3) Changes in the Su-PS skin reaction provided an index for evaluating the therapeutic effects of OK-432 administration in patients with gastric cancer. These results suggest that the Su-PS and Su-PR skin reaction tests are important in the treatment of patients with gastric cancer.

Advanced ovarian cancer treated by intraperitoneal immunotherapy with OK-432.

Twelve patients with advanced ovarian cancer with ascites were treated by intraperitoneal immunotherapy with OK-432 in combination with surgery and systemic immunochemotherapy. The median time for total resolution of ascites after the OK-432 injection was 7.1 days. In three of the four patients who died, it took more than the average number of days for the ascites to disappear. The ascites index, the ratio of the ascitic fluid volumes before and after intraperitoneal immunotherapy, was significantly higher in the patients who died than in those who survived. The side effects of intraperitoneal administration of OK-432 varied, but were not serious and relief was not needed in most cases. Intraperitoneal injection of OK-432 was considered effective in preventing the reappearance of ascites in these cases of advanced ovarian cancer, and the ascites index was believed to be related to the prognosis.

Streptococcal preparation (OK-432) inhibits development of type I diabetes in NOD mice.

OK-432 (a streptococcal preparation) has been widely used for cancer immunotherapy in Japan. It is the most potent immunomodulator in activating both macrophages and killer T cells and in increasing interleukin 2 production. Two K.E. (Klinische Einheit, clinical unit) of OK-432 were given intraperitoneally to each of 17 female nonobese diabetic (NOD) mice every week from 4-24 wk of age. NOD mice as well as BB rats spontaneously develop type I diabetes. During administration of OK-432, the development of diabetes was inhibited in 17 of 17 mice over the 24-wk observation period, whereas 14 of 17 female NOD mice given physiological saline had developed diabetes by 24 wk of age. At the onset of diabetes, nonfasting blood glucose was 511 +/- 82 mg/dl. Histologic examination showed that in the OK-432-treated NOD mice, 98% of total islets were intact or mildly infiltrated with mononuclear cells, whereas in saline-treated NOD mice, 79% of total islets exhibited severe insulitis. In OK-432-treated NOD mice, both the number of the mononuclear spleen cells and their natural killer cell activity was significantly increased.

Treatment of pleuritis carcinomatosa of primary lung cancer with intrapleural administration of OK-432

Treatment of pleuritis carcinomatosa of primary lung cancer with intrapleural administration of OK-432

Correlation between the presence of intracellular OK-432 and antitumor activity of peritoneal macrophages

The mechanism of the generation of tumoricidal macrophages by intraperitoneal administration of OK-432 was investigated. An association between the presence of intracellular FITC-conjugated OK-432 and the increase of macrophage cytostatic activity was observed after i.p. administration of OK-432. When peritoneal exudate cells were fractionated on the basis of cell size by velocity sedimentation, a higher proportion of macrophages and a strong cytostatic activity were shown in the large cell fractions. Furthermore, in these fractions, increased numbers of macrophages with intracellular FITC conjugated and 3H-acetate labelled OK-432 were observed. Thus, the more cytocidal macrophages can be identified by the presence of intracellular microorganisms. A role for OK-432 in the direct activation of macrophages is suggested.

上顎洞に発生した腺様嚢胞癌に対する集学的治療

The most effective treatment for adenoid cystic carcinomas arising from minor salivary glands is surgical removal of primary lesion including the wide safety margin. However, it is well known that the salivary gland tumors have poor prognosis and high frequency of metastasis. Therefore, the multidisciplinary therapy that prevents effectively recurrence and metastasis is needed.Two cases of adenoid cystic carcinoma of the maxillary sinus were treated according to the follwing multidisciplinary method. Case 1 (T3NOMO by JJC proposal): The patient had surgical removal of the affected maxilla and then radiation therapy. Four weeks later, adjuvant chemotherapy (Adriamycin and Cisplatin) was performed. In addition, UFT and OK-432 were administered continuously up to date from the onset of the treatment. Consequently, the patient remains be disease-free 24 months postoperation. Case 2 (T4NOMO by JJC proposeal): The patient was treated with the multidisciplinary methods such as surgical reduction of a tumor-burden, radiation therapy, intraarterial infusion of cisplatin and 5-FU into a superficial temporal artery and intradermal administration of OK-432. Thereafter, complete regression of the primary tumor was observed. However, metastasis was detected in the regional lymph nodes, which were subjected to radical neck dissection. The patient remains be disease free 8 months postoperation. These findings indicate that the multidisciplinary therapy may improve the prognosis of adenoid cystic carcinoma of the maxillary sinus.

Acute liver failure in rats inhibited by intrasplenic administration of OK-432

Intrasplenic administration of OK-432, an immunostimulant derived from Streptococcus, prevented hepatic failure induced in rats by d-galactosamine. When OK-432 was given 1.0 K.E. (Group I) or 0.1 K.E. (Group II) into the subcutaneously transpositioned spleen three times prior to dosing with d-galactosamine, survival rates were 100 and 87%, respectively. On the contrary, with a splenic injection of saline (Group III), the survival rate was 47 and 32% in rats given OK-432 1.0 K.E. intraperitoneally (Group IV). The poisoned rats given no pretreatment (Group V) survived at a rate of 26%. These results show that intrasplenic administration of OK-432 leads to a significant enhancement of survival. Metabolic data and histological findings were compatible with survival rates, in each group. Activation of the reticuloendothelial function by the intrasplenic administration of this immunostimulant seems to have prevented acute liver failure.

Antitumor effect in peritoneal carcinomatosis of intraperitoneal administration of a streptococcal preparation, OK-432: I. Experimental study in the rat.

The effect of intraperitoneal OK-432 administration and its mechanism of action were investigated using an experimental rat tumor. On day 4 or 6 of intraperitoneal inoculation of tumor cells, 50 Klinische Einheit (KE)/kg of OK-432 was intraperitoneally administered, and intraperitoneal exudate cells were microscopically examined daily. Distribution of peritoneal exudate cells: A marked increase in neutrophils was noted in ascites at 24 hours after intraperitoneal administration of OK-432, with macrophages and lymphocytes appearing later than neutrophils. A subsequent decrease in neutrophils was accompanied by a gradual increase in tumor cells. Findings of tumor cells: When OK-432 was intraperitoneally administered on day 4 of tumor inoculation, aggregation of neutrophils around tumor cells was observed on day 6, and rosette formations around tumor cells were noted on day 7 or 8. Subsequently tumor cells that formed the centers of the rosettes were destroyed and disappeared. These reactions were not observed when OK-432 was administered on day 6, nor were they observed in untreated cases. These results suggest that cytotoxic effects of intraperitoneally administered OK-432 were displayed mainly by rosette formation by neutrophilic cells, as far as this model was concerned.

Effects of intraperitoneal administration of OK-432 for patients with advanced cancer.

The effects of intraperitoneal administration of OK-432 on tumor cells in ascites, in relation to the infiltration of effector cells and on the immune responses of the host, particularly, with regard to immune suppressive mechanisms, were investigated in 25 patients with cancerous ascites. The effects of OK-432 depended on frequency of the repeated and continuous administrations through a tube placed in the peritoneum during laparotomy. Infiltrations of neutrophils and lymphocytes were observed in the ascites within a short period after the administration and monocyte infiltration followed. Disappearance of tumor cells correlated well with the infiltration of these cells. No marked changes in the proliferative responses of peripheral blood lymphocytes were noted and decreases in serum inhibitory factor levels in sera were observed in patients given larger doses of OK-432. A marked reduction in Concanavalin-A-induced suppressor cell activities was observed after OK-432 administration. OK-432 administration probably leads to a disappearance of tumor cells by enhancing peritoneal effector cell activities and by inhibiting the induction of suppressor cell activities, in a dose dependent manner.

The effects of oral administration of streptococcal preparation OK-432 (picibanil) on the immunity of mice with transplanted cecal tumors

The effects of oral administration of streptococcal preparation OK-432 (picibanil) on the immunity of mice with transplanted cecal tumors

急性白血病寛解導入療法に関する研究

The streptococcal preparation, OK-432, has been found to be a biological response modifier. The effects of OK-432 on autologous CFUs in mice and hematopoietic disorders induced by anti-leukemic agents (DNR: daunorubicin, ara-C: cytosine arabinoside, NCS: neocarzinostatin) were investigated in order to obtain information as to the possibility of its clinical application in induction chemotherapy for acute leukemia.OK-432 increased autologous CFUs in irradiated mice, suggesting that it acted hematopoietically on multipotent stem cells. In mice treated with DNR, OK-432 reduced neutropenia when it was given before and simultaneously with DNR. In mice pretreated with OK-432, the duration of neutropenia caused by ara-C tended to be prolonged. On the other hand, simultaneous administration of OK-432 with ara-C showed no significant differences as compared to ara-C alone.These results suggested that the simultaneous administration of OK-432 with chemotherapeutic agents might protect normal hematopoiesis during induction chemotherapy for acute leukemia in man.

Oral administration of OK-432 induces human natural killer like cells in patients with malignant solid tumor

Oral administration of OK-432 induces human natural killer like cells in patients with malignant solid tumor

Immune responses to an oral administration of OK-432 in the aged subjects without malignancy

OK-432, one of the so-called biological response modifiers (BRMs), has been reported to restore cellular and humoral immune responses in tumor-bearing individuals. However, since host immune responses in tumor-bearing individuals are modified by development of disease, by application of various forms of anticancer therapy, surgical operation, and by radiation therapy, it is sometimes impossible to evaluate accurately the effect of these nonspecific BRM restoring impaired immune response in patients with malignancy. Avoiding this problem, we studied the effect of BRM on host immune responses in the aged individuals without malignancy, who had shown decreased cellular immunity detected by PPD and PHA skin tests and in vitro blastoid transformation test of lymphocytes by PHA. Although OK-432 is used widely by intradermal or intramuscular injections, it is more convenient for the patients to receive oral administration than intramuscular injection having the side effects such as chill and fever.Five KE or 20 KE of OK-432 was orally given three times a week for 1 month to the aged patients without malignancy who showed decreased skin responses to PPD and PHA. Oral adminstration of 5 KE of OK-432 evoked an increased skin response to PPD except 2 patients. PPD skin response was much more increased by the oral administration of 20 KE of OK-432, and the increased responsiveness was detected even 2 months after withdrawal of OK-432. There was same tendency in the skin response to PHA. The increased skin reaction to SuPS (lipopolysaccharide fraction of Streptococcus pyogenes Su strain) was observed in the one group of patients treated with 5 KE of OK-432, while there was no significant increased response by oral administration of 20 KE of OK-432.It was concluded that the activation of non-specific host immune responses assessed by the skin tests used by us could be evoked by the oral administration of OK-432 as well as intramuscular injection.

Augmentation of interleukin 1 and interleukin 2 production by OK-432.

Intraperitoneal (i.p.) administration of OK-432 augmented both interleukin 1 (IL-1) and interleukin 2 (IL-2) production to the rechallenge of OK-432 in vitro. Peritoneal exudate cells (PEC) of mice 8 days after i.p. injection with OK-432 (1 KE/mouse) showed maximum IL-1 production to the restimulation with OK-432 in vitro. OK-432-induced IL-1 was consisted of three molecular weight species (two major peaks: 85 K and 15 K daltons and one minor peak: 67 K daltons) on Sephadex G-100 chromatography. Splenocytes of mice 4 days after i.p. injection with OK-432 (1 KE/mouse) demonstrated maximum IL-2 production to the in vitro rechallenge of OK-432, however, in vivo OK-432 administration failed to enhance ConA-induced IL-2 production in vitro. From gel filtration analysis, OK-432 induced IL-2 had an unique molecular weight (approximately 70 K daltons). From these results, OK-432-induced augmentation of cellular immunity against tumor cells might be due to the activation of so-called lymphokine cascade reaction mediated by IL-1 and IL-2.

Intrapleural administration of OK432 in cancer patients: augmentation of autologous tumor killing activity of tumor-associated large granular lymphocytes.

Ten patients with carcinomatous pleural effusions were treated with single intrapleural (i.pl.) injections of the streptococcal preparation OK432 on day 0 and the effects of i.pl. OK432 on the lysis of fresh or cryopreserved autologous tumor cells isolated from the pleural effusions were observed on day 7. In eight patients tumor cells in the effusions had decreased or disappeared by day 7. The other two patients, however, had no clinical evidence of therapeutic benefit from i.pl. OK432. Effusion tumor cells were relatively resistant to lysis by autologous lymphocytes when tested in a 4-h 51Cr-release assay. Positive reactions were recorded for blood and effusion lymphocytes in two of ten untreated patients. Injection of OK432 i.pl. resulted in an induction or augmentation of cytotoxicity against autologous tumor cells and K562 in the effusions of seven of ten subjects by day 7. In contrast, autologous tumor killing activity of blood lymphocytes was not always modified by i.pl. OK432. Purification of large granular lymphocytes (LGL) by discontinuous Percoll gradient centrifugation enriched autologous tumor killing activity, with no reactivity in LGL-depleted, small T lymphocytes. Significant lysis of autologous tumor cells was observed with effusion LGL from seven of ten untreated patients. Seven days after i.pl. OK432 injection, effusion LGL expressed enhanced cytotoxicity against autologous effusion tumor cells, whereas T cells were still not cytotoxic to autologous tumor cells on day 7. The frequency of LGL among effusion lymphocytes was not altered by i.pl. OK432. Adherent effusion cells were not involved in lysis of autologous effusion tumor cells in either untreated or OK432-treated patients. In vitro treatment of blood and effusion lymphocytes with OK432 induced an enhancement of autologous tumor-killing activity in patients who subsequently responded to i.pl. OK432 treatment. OK432 augmented in vitro autologous tumor killing activity of LGL, whereas T cells failed to lyse autologous tumor cells even after in vitro activation with OK432. These results indicate that i.pl. administration of OK432 to cancer patients will result in an augmentation of autologous tumor killing activity of LGL in the pleural effusions, and that this could be responsible for the antitumor activity of i.pl. OK432 therapy.

Effects of intradermal administration of streptococcal preparation OK-432 on interferon and natural killer cell activities in patients with oral cancer

The streptococcal preparation OK-432 was used by intradermal administration as an immunotherapy in 18 patients with oral cancer, and the sera from patients during OK-432 treatment were serially assayed for interferon (IFN) activity by the plaque-reduction method with vesicular stomatitis virus in FL cells derived from human amniotic membrane. The type of serum IFN was characterized by acid-treatment and neutralization test with anti-IFN-α and anti-IFNβ antisera. IFN-γ was expressed for its titer as the residual IFN activity after neutralization with both antisera. An intradermal injection of OK-432 transiently induced IFN activity and 3 patterns in the type and level of the produced IFN were observed. Although most of the patients induced IFN-γ and acid-stable IFN or only IFN-γ, 2 patients seemed to be unresponsive to OK-432. When we examined the relationship between natural killer (NK) activity and IFN titer, a sharply declined NK activity was found immediately post OK-432 administration, and then NK activity stayed around the pretreat-ment level. Most of the tested patients' induced IFN-γ, preceding the step toward the gradual increase in NK activity, decreased with OK-432. However, even in the patients showing no IFN induction with OK-432, a significant decrease of NK activity occurred.

Activation of macrophage function by intraperitoneal administration of the streptococcal antitumor agent OK-432

Motility, adhesiveness, IL1 production, and inhibition of tumor cell growth in vitro were examined in murine peritoneal macrophages obtained after intraperitoneal injection of a streptococcal preparation OK-432, heat-inactivated OK-432 (HI-432), and thioglycollate medium (TG). By varying the interval between intraperitoneal injection of OK-432 and the harvest of peritoneal macrophages, it was found that OK-432 induced a time-dependent multi-step alteration of these properties: step I increased motility on day 1; step II increased adhesiveness on day 2; and step III increased inhibition of tumor cell growth and IL1 production. During step III, the peritoneal macrophage population, including Ia-bearing cells, increased dramatically in the peritoneal cavities of OK-432-treated mice. In contrast, injection of either HI-OK-432 or TG, which lack antitumor activity in vivo, initiated steps I and II, but not step III. The Ia-bearing macrophages induced by OK-432 showed high ability of IL1 production, but low growth inhibitory activity against tumor cells. Based on these results, OK-432 seems to be performing a dual function: eliciting a new population of macrophages to the site of injection (heat stable function), and inducing two different populations of antitumor macrophages and Ia-bearing macrophages (heat unstable function).

Intrapleural administration of OK432 in cancer patients: activation of NK cells and reduction of suppressor cells.

Twelve patients with carcinomatous pleural effusions were treated with single intrapleural (i.pl.) administration of OK432 on day 0 and the effects of i.pl. OK432 on natural killer (NK) cell activity were followed on day 7. Two patients showed no clinical evidence of therapeutic benefit from i.pl. OK432. In the other 10 patients, pleural effusions and/or tumor cells in the effusions had decreased or disappeared by day 7. NK cell activity was markedly low or absent in pleural effusions of untreated patients due to the presence of adherent effusion cells capable of suppressing the maintenance and interferon-induced augmentation of NK cell activity. I.pl. injection of OK432 resulted in enhancement of NK cell activity and abrogation of NK suppressor cell activity in the effusions. On the other hand, blood NK cell activity was not consistently altered by i.pl. OK432. In vitro treatment of effusion mononuclear cells from untreated patients with OK432, but not with interferon, augmented NK cell activity. In addition, adherent effusion cells of untreated patients lost their NK suppressor function following in vitro OK432 treatment. These results suggest that i.pl. administration of OK432 will result in augmentation of NK cell activity and reduction of NK suppressor cell activity in pleural effusions, which could be responsible for the antitumor activity of i.pl. OK432.

肺癌の免疫化学療法

The functions of peripheral blood monocytes in 14 untreated patients with lung cancer were examined before and after a 1-week course of OK-432 therapy(1 KE every day).The absolute monocyte count was calculated from the percent of monocytes in the total leukocyte count. The mean absolute numbers of monocytes before and after administration of OK-432 were 400/cmm and 437/cmm, respectively though the difference was not statistically significant.The lysosomal enzyme activity of monocytes was histochemically determined by beta-galactosidase staining. The mean percentages of beta-galactoside positive monocytes before and after administration of OK-432 were 31.2 and 47.7 respectively. The increase was significant (P<0.05).The monocyte chemotactic response was measured in vitro by Boyden's millipore technique using zymosan activated human serum as the chemoattractant. The monocyte chemotactic responses before and after administration of OK-432 were 20.1 and 25.2, respectively and a significant difference was found between the two groups(P<0.01).The ability of monocytes to phagocytose complemented zymosan was accelerated by OK-432 administration, but the acceleration was not statistically significant.Cytostatic activities of monocytes on EBC-1 target cells before and after administration of OK-432 were 84.5 and 91.4, respectively the difference was significant (P<0.05).These results indicate that OK-432 activates the reduced monocyte functions in lung cancer patients in vivo, and that OK-432 may be a useful immunotherapeutic agent in man.