In vitro anti-tumour activity of tumour necrosis serum (original) (raw)
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Cancer Research, 1987
Thymostimulin (TS), a partially purified thymic factor, has a signifi cant impact on tumor development in C57B1/6 mice inoculated with Lewis lung carcinoma (3LL) cells, as judged by its effect on time of tumor appearance after tumor cell transplantation. In a previous study, we determined the conditions under which survival rate of the tumor-bearing mice can be significantly increased by TS treatment. In the study com municated here we analyzed host defense mechanisms that are modified by TS treatment in the tumor-bearing mice. In general, immune param eters that were increased or stimulated by the presence of the tumor were further increased in the TS-treated animals (number of lymphoid spleen cells, their response in mixed lymphocyte tumor cultures, their natural killer cell activity, and their ability to produce colony-stimulating factor), or reached earlier maximum levels (spontaneous [3H|thymidine incorpo ration, a reflection of in vivo spleen cell activation). Responses which reflect tumor-induced immunosuppression (proliferative response induced by phytohemagglutinin or concanavalin A stimulation) were restored to normal level by TS. Specific tumor-related reactions (specific cell-medi ated cytotoxicity) were preserved in the TS-treated animals. The wide spectrum of TS effects had, nevertheless, certain elements of selectivity; e.g. colony-stimulating factor, but no Interferon production is enhanced by TS in the tumor-bearing mice in diametric contrast to TS effect in Mengo virus-infected mice. The spectrum of TS effects was also depend ent on the type of tumor cell used. The results indicate that the significant effect of TS on 3LL tumor development in mice is associated with a strong, multifaceted effect of TS on the immune system.
II. Bacterial Immunity and Tumor Growth; Effect of Bacterial Products on Tumors Milton
2011
The effect of B. danysz on the Ehrlich mouse carcinoma was described in a previous report (1936). Hyperemia and other changes, which were interpreted as being of a regressive nature, were observed in the tumors following intratumoral injection of this highly pathogenic microorganism. The experiments to be described in this paper deal with the attempt to immunize mice against infection with B. danysz by inoculation of a vaccine of that organism, the study of tumor transplants in these animals, and the reaction of the animals as well as the tumors to the injection of virulent B. danysz as compared to that of immunized but non-tumor-bearing animals. The effect of intraperitoneal injections of toxin and heat-killed cultures of B. danysz on the Ehrlich mouse carcinoma was also studied. Torrey and Kahn (1927), during their studies of the effect of proteolytic anaerobes on tumors, attempted to immunize rats actively and passively against B. histolyticus. They then noted the effect of this ...
Biochemical studies of cytokinetic changes during tumor growth
Cancer research, 1970
larger after irradiation or chemotherapy than would be pre dicted from cell survival curves alone. In this paper we examine some of the biochemical factors that may contribute to the cytokinetic changes which charac terize the growth of a hyperdiploid Ehrlich ascites tumor in vivo. Although the rate of protein synthesis declines during growth of this tumor, there is no evidence for qualitative defects which might cause the cytokinetic changes. Our results support the view that accumulation of cells in G1 during tumor growth reflects the progressively larger proportion of anoxic cells in the population. MATERIALS AND METHODS Hyperdiploid Ehrlich ascites tumor cells (iO@) were inocu lated i.p. into female Swiss-Webster mice (20 to 25 g). The growth characteristics of this tumor are described elsewhere (30). Unwashed cells were collected aseptically, suspended in their own ascitic fluid, and diluted with 0.9% NaCl solution as required for inoculation. When tumor enzyme activity was to be assayed on the 1st day ofgrowth, the prospective recipient mice were exposed to 1000 R X-radiation immediately before tumor inoculation to reduce the proportion of small lympho cytes to the level (3 to 10%) found in established tumors (30). Cells were enumerated with a Model B Coulter counter. Imposition of Hypoxia In experiments designed to test the effects of hypoxia on tumor growth, mice bearing the Ehrlich ascites tumor were placed in a Lucite chamber which was flushed with a mixture of 6 or 9% oxygen in nitrogen. The oxygen content of the effluent gas was monitored at frequent intervals with a Beck man oxygen analyzer. Food and water were available ad libitum. Measurement of Cell Cycle Parameters The distribution of cells within the cell cycle was measured by the labeled mitosis method. TdR-3 H (0.36 Ci/mmole, 12 @.zCi/mouse) in 0. 15 ml or 0.30 ml of 0.9% NaCl solution was injected i.p. and 2 or 3 mice were sacrificed at appropriate intervals. The thymidine index (proportion of cells in S) was determined by sacrificing animals 20 ruin after injection of TdR-3 H. For the â€oecontinuouslabeling―experiment, TdR-3 H was injected i.p. every 4, 6, or 8 hr. In all cases, the cells were SUMMARY We have measured the cytokinetic changes which charac terize the growth of a hyperdiploid Ehrlich ascites tumor and have investigated some possible underlying biochemical mechanisms. In this tumor, the cell cycle increases from 9.5 hr on the 2nd day of growth to 39 hr on the 14th day and the proportion of cells in S (thyrnidine index) declines from 80 to 40% during the same interval of growth. Protein synthesis (measured by uptake of leucine-' 4C during a 20-ruin pulse) declines at a rate similar to the decline of the thymidine index, but there are no changes in the specific activity of six en zymes: aryl sulfatase, @3-glucuronidase, thymidine kinase, glucose-6-phosphate dehydrogenase, alkaline phosphatase, and glutathione reductase. Transplantation of l0'@ cells from plateau-phase tumors (14 to 16 days) to new hosts causes the Tumor rate of protein synthesis to increase immediately. The thy midine index, on the other hand, remains constant for 5 to 6 hr after transplantation and then doubles within the next 4 hr. The rise is prevented (or delayed) whep the animals breathe 6% oxygen. Hypoxia also decreases the thyrnidine index of 2-day tumors, but it does not alter the length of the cell cycle. The results suggest that the progressive accumulation of cells in G1 during tumor growth may reflect the relative ineffi ciency of anaerobic metabolism in producing the energy (e.g., ATP) necessary for movement into S phase. 1 Work performed under the auspices of the United States Atomic Energy Commission.
Cancer Immunology Immunotherapy, 1992
The effect of prothymosin o~ (ProTon) on the survival of DBA/2 mice inoculated with syngeneic tumour cells was studied. DBA/2 mice inoculated intraperitoneally (i.p.) with 2x105 syngeneic leukaemic L1210 cells developed ascites within 8-12 days and died 10-14 days later. Treatment with ProTc~ consistently inhibited the development of ascites in 20% of the treated animals and prolonged the survival of 40%-60% of the animals up to 70 days. The most effective treatment schedule of ProTcz was 300 ng/mouse given i.p. at 2-day intervals for 3 weeks followed by a rest period of 7 days, prior to tumour cell inoculation. Peritoneal exudate (PE) cells collected from mice treated with the optimal dose of ProT(z produced, in the absence of exogenous stimulus, six-to eightfold higher levels of tumour necrosis factor o~ (TNFc~) than PE cells from control mice. Furthermore these cells exhibited cytotoxic activity against several tumour cell lines including the syngeneic L1210, the TNF-insensitive P815 mastocytoma, the human MOLT-4 lymphoblastic leukaemia, as well as the murine TNF-sensitive L929 fibroblast cell line. Kinetic studies revealed that both production of TNFcz and tumoricidal activity peaked 7 days after the last injection of ProT~ and were maintained at high levels over a period of 1 month: Injections with 150 ng ProTo~ slightly improved the survival of mice whereas higher (500 ng and 1000 ng) doses of ProT~ and a wide range of thymosin czl doses remained without any effect. PE cells collected from these mice produced extremely low levels of TNFcx and exhibited negligible tumoricidal activity. Our data demonstrate that ProT(z has a protective effect in vivo against the growth of adoptively transfered tumour cells and suggest that this effect is, at least in part, mediated by ProT(~-activated PE cells. These cells were demonstrated to produce high levels of TNFa in vitro and to exhibit activity against both TNF-sensitive and TNF-resistant cell lines.
Cancer Immunol Immunother, 1992
The effect of prothymosin o~ (ProTon) on the survival of DBA/2 mice inoculated with syngeneic tumour cells was studied. DBA/2 mice inoculated intraperitoneally (i.p.) with 2x105 syngeneic leukaemic L1210 cells developed ascites within 8-12 days and died 10-14 days later. Treatment with ProTc~ consistently inhibited the development of ascites in 20% of the treated animals and prolonged the survival of 40%-60% of the animals up to 70 days. The most effective treatment schedule of ProTcz was 300 ng/mouse given i.p. at 2-day intervals for 3 weeks followed by a rest period of 7 days, prior to tumour cell inoculation. Peritoneal exudate (PE) cells collected from mice treated with the optimal dose of ProT(z produced, in the absence of exogenous stimulus, six-to eightfold higher levels of tumour necrosis factor o~ (TNFc~) than PE cells from control mice. Furthermore these cells exhibited cytotoxic activity against several tumour cell lines including the syngeneic L1210, the TNF-insensitive P815 mastocytoma, the human MOLT-4 lymphoblastic leukaemia, as well as the murine TNF-sensitive L929 fibroblast cell line. Kinetic studies revealed that both production of TNFcz and tumoricidal activity peaked 7 days after the last injection of ProT~ and were maintained at high levels over a period of 1 month: Injections with 150 ng ProTo~ slightly improved the survival of mice whereas higher (500 ng and 1000 ng) doses of ProT~ and a wide range of thymosin czl doses remained without any effect. PE cells collected from these mice produced extremely low levels of TNFcx and exhibited negligible tumoricidal activity. Our data demonstrate that ProT(z has a protective effect in vivo against the growth of adoptively transfered tumour cells and suggest that this effect is, at least in part, mediated by ProT(~-activated PE cells. These cells were demonstrated to produce high levels of TNFa in vitro and to exhibit activity against both TNF-sensitive and TNF-resistant cell lines.
Breast Cancer Research and Treatment, 1993
A detailed analysis of the immune system response has been performed during the development and progression of dimethylbenz(a)anthracene (DMBA)-induced rat mammary tumors. For this aim, a number of immune parameters (thymocyte and splenocyte proliferative response to T-dependent mitogens, antibody production, lymphocyte subset phenotyping, interleukin 2 receptor expression in resting and activated lymphocytes, thymus morphology and morphometry), were correlated with tumor appearance and growth at different (- 7, 0, + 15, + 30, + 60, + 90, and + 120 days) time intervals after intragastric administration of DMBA, in the absence or the presence of a concomitant treatment with the thymic pentapeptide thymopentin (TP5). A profound and time-dependent immunosuppression characterized the treatment with the carcinogen. Both cell-mediated and humoral immune responses showed a 50% inhibition 2 weeks after DMBA administration, with a peak after 30 days, followed by a plateau until 120 days of observation. The mechanism responsible for reduced ability of thymocytes and splenocytes to respond to both Con-A and PHA was explained by the significant inhibition of one of the key steps of T cell activation, namely the expression of IL-2 receptor in lymphocytes from DMBA-treated animals. The flow cytometric analysis of lymphocyte subpopulations revealed an important reduction in the overall populations of thymocytes and splenocytes. At the thymus gland level, a dramatic reduction of double positive CD4+CD8+ and a decrease of CD4+CD8- and CD4-CD8+ were observed, together with a marked atrophy of the thymic cortex, and impairment of the thymic microenvironment. One hundred and twenty days after DMBA administration, approximately 60 to 70% of the animals developed tumors with a mean tumor surface area of 2.88 ± 0.86 cm2, and a number of 2.44 ± 1.0. Treatment with TP5 (100 ng/animal, three times a week, starting a week before DMBA), produced specific effects on different immune compartments and tumoral growth, characterized by a significant reversal of immune depression with a stimulatory effect measured on lymphoproliferative assays, lymphocyte subset distribution, and IL-2 receptor expression. Moreover, thymic atrophy was almost completely prevented in TP5 treated animals. Of major interest, a significant delay in the appearance and growth of tumors was observed in TP5 treated rats. When DMBA-treated animals were followed for the entire observation period (0–120 days) and the immune responsiveness correlated according to tumor progression, stability, or regression, a positive correlation was calculated between the degree of immune system depression and the individual rate of tumor growth; in TP5-treated rats the majority of the tumors were static or regressing tumors. It is concluded that a profound and specific suppression of thymus-dependent immune functions follows the treatment with the carcinogen DMBA. Such immune deficits appear 2 weeks after the carcinogen administration, and may represent early indicators of tumor development and growth. A key role is suggested for the thymus gland and its peptide secretion, since the dramatic atrophy of this gland and the abnormal thymusdependent immune parameters studied are almost completely reversed by a synchronized treatment with thymopentin. Although from the present results it seems not possible to conclusively assign a specific causeeffect relationship, from the parallel delay in tumor appearance and growth in TP5-treated rats it seems tempting to speculate that the decline in thymus-dependent immune functions may play a major role in tumor development.
Antitumor activity with nontoxic doses of protein A
Cancer Immunology Immunotherapy, 1984
This report confirms our previous observation that 1V inoculation of purified protein A causes regression of rat mammary adenocarcinomas. In treated tumors, we have obtained histological evidence of changes indicating tumor cell destruction. Protein A treatment does not cause reduction in the body weight or organ weights of rats; nor does it cause any decrease in activity of the enzymes of the microsomal mixed function oxidase system in the liver. Protein A stimulates peripheral white cell counts in normal rats, but not in tumor-bearing rats. We found that protein A infusion reduced (P < O. 0005) the level of circulating plasma immune complex concentration. A homing study with lZ~I-labeled protein A indicated that liver, spleen, and kidney tissues are the major sites of protein A accumulation. Therefore, protein A seemed to exert its antitumor effects without causing any generalized toxicity to the system. It is postulated that the action of protein A may be related to its ability to cause a drastic reduction in circulating plasma immune complex concentration, thus potentiating the immune reactivity of the host" observed earlier.