Ascites tumor production

Monoclonal antibodies can be produced not only in a cell culture but also in live animals. When injected into mice (in the peritoneal cavity, the gut), the hybridoma cells produce tumors containing an antibody-rich fluid called ascites fluid. Production in cell culture is usually preferred, as the ascites technique may be very painful to the animal and if replacement techniques exist, may be considered unethical. The process of producing monoclonal antibodies described above was invented by Georges Kohler. Cesar Milstein, and Niels Kaj Jeme in 1975 they shared the Nobel Prize in Physiology or Medicine in 1984 for the discovery (http //en.wikipedia.org/ wiki/Antibody). 

Pannell, R. and Milstein, C. (1992) An oscillating bubble chamber for laboratory scale production of monoclonal antibodies as an alternative to ascitic tumors J Immunol Methods 146,43—48... 

W15. Wong, C. K., Fung, K. P., Lee, C. Y., and Choy, Y. M., In vivo production of tumor necrosis factor for the treatment of mice bearing Ehrlich ascites tumor. Cancer Lett. 63,7-13 (1992). 

Unlike the production of other proteins from animal cells, mAbs can also be produced in vivo by inducing ascitic tumors in laboratory animals,... 

With Potter s development of experimental plasmacytoma in mice, it was established that the turnover of paraprotein (N2), or more simply the serum level (08), was directly related to the weight of solid soft-tissue plasmacytoma. In our laboratory an ascitic form of plasmacytoma has been studied, and using isotope dilution it has been possible to estimate the actual total number of plasmacytoma cells in a mouse. At the same time the serum level of paraprotein was measured and a simple correlation was shown (F2). To the best of my knowledge, this was the first time that the serum level of a tumor product had been directly related to the actually counted number of tumor cells. Incidentally it was noted that the paraprotein could be first detected in the serum, when a 23 g mouse had 3 million tumor cells. 

Rats are also suitable for the production of monoclonal antibodies on a 10-times larger scale (Galfre et al., 1979). The Y3 myeloma (K-producing) of Lou rats can be used and ascites tumors can be obtained two weeks after an i.p. injection of 0.5 ml pristane by the i.p. injection of 5 x 10 hybridoma cells. A minor disadvantage is the unreactiveness of rat IgG2a with protein A (Rousseaux et al., 1981). The use of rabbit spleen cells with mouse myelomas has not yet been successful. 

Production of ascites tumors About 10 cells from... 

Calcium chromate has been shown to induce cytoplasmic petite mutations in mitochondria of Saccharomyces cerevisiae K Calcium chromate also dramatically depressed the content of the mitochondrial gene products cytochrome aa3 and cytochrome b, in whole yeast cells. Chromate ( 8 nM) was readily taken up by rat thymocytes and after 30 min 9% of the Cr was found in the mitochondria although 62% was found in the nuclei . Isolated rat thymus mitochondria and nuclei readily took up CrOj . After one hour incubation of Erlich ascites tumor cells with CrOj (380 /nuclear fraction and 12% was in the mitochondrial-microsomal fraction. Levels of chromium in rat liver mitochondria reached a plateau six hours after i.v. injection of chromate (0.02 mg/kg) and remained at that level through 5 days. Liver nuclear chromium levels in the same animals, although similar to mitochondrial levels at 6 h, reached a maximum at 12 h and steadily decreased after that time. Therefore the nuclear chromium levels were lower than the mitochondrial chromium levels at later times (24-120 h) after injection. The subcellular distribution of chromium in the liver of rats injected i.v. with chromate (0.56 mg/kg) was also found to be time dependent in another study. The distribution of chromium in rat liver mitochondria increased from 5% at 15 min to 21% at 72 h and also increased in the nuclear fraction from 22% at 15 min to 52% at 72 h. Incubation of isolated rat liver mitochondria with chromate (0.3-16.6 electron transport chain of the mitochondrial iner membrane. 

Figure 2 includes an additional, parallel route for the decomposition of Ii, as revealed by the distinctive EPR signature of the final product at pH 5. We proposed it to be also a dinitrosyl species, [Fe(CN)2(NO)2] (I2). new member of the well-characterized series of paramagnetic distorted tetrahedral complexes with different L ligands, described as Fe(NO)2 - These species behave as reversible, labile NO carriers, involved in trans-nitrosylation processes. EPR signals ass nable to these dinitrosyl complexes have been found in tissue of ascite tumors of mice upon injection with SNP. Those containing L = thiolates and imidazole were found to activate sGC promoting vasodilation (140). 

Another method for producing hybridomas is as ascites tumors in mice. In this method, hybridomas are injected into animals. The antibody is contained in the rich ascites fluid that develops. Large doses of cells are required to initiate the tumor and it is important that the hybridoma and mouse are compatible. The productions of ascites tumors are facilitated by pretreatment of the mouse with, and intraperito-neal injection of, pristane. The ascites fluid is drained once sufficient swelling has been produced at the tumor site. The main concerns with this method are animal welfare issues, but the main advantage is that large amounts of antibody can be prepared. 

In cell lines such as mouse myeloma or Ehrlich ascites tumor cells some 20% of the total cytoplasmic mRNA is not associated with ribosomes but exists in an untranslated state even though deproteinised mRNA prepared from these mRNP particles is perfectly translatable m vitro. A more or less trivial explanation for such untranslated mRNA is that it originates from a small fraction of the cells in which the rate of initiation is very low, e.g. cells in mitosis. A comparison of the products of vitro translation of this mRNA and of the polysomal mRNA renders this explanation less than wholly satisfactory. In extreme cases, the non-translated mRNPs are found to contain an entirely different set of mRNA species from... 

Grazianl, Y., Wlnlkoff, J. and Chayoth, R. (1977). Regulation of cyclic AMP level and lactic acid production In Erlich ascites tumor cells. Blochim. Blophvs. Acta 497. 499-506. 

Podhajcer, O.L., Friedlander, M. and Graziani,Y. (1980). Effect of liposome-encapsulated quercetin on DNA synthesis, lactate production, and cyclic adenosine 3 5 -monophosphate level in Erlich ascites tumor cells. Cancer Res. 40, 1344-1350. 

The product of ribonuclease action is an amino acid derivative of adenosine. This was first obtained from amino acid-RNA compounds of rat liver by Zachau, Acs, and Lipmann (187), then from ascites tumor cells by Hecht et al. (193), and from E. coli by Preiss et al. (194). 

This reaction has been applied to the elaboration of steroid skeletons and to sarkomycin (a natural product which shows inhibitory effect on Ehrlich ascites tumor) synthesis. Coronafacic acid synthesis is another interesting example of a palladium-catalyzed cyclization. 

Isolated polypeptides, viscotoxins II, III, and IVb, have been associated with cardio-toxicity but have also been found to exhibit cytotoxic activity against human tumor cells of the KB and HeLa lines in tissue culture. A peptide with a molecular weight of 5000 was found to be cytotoxic to Dalton s lymphoma ascites tumor cells in vitro in mice, without affecting normal lymphocjfies, indicating a cell-dependent specificity also cyctotoxic to Ehrlich ascites cells, both prophylacticaUy and after tumor development. Commercial mistletoe products have been used to treat various cancers in Europe with clinical success. A group of 50 cases of carcinomatous pleural effusions were treated with a topical preparation for an average of 3.3 application over 18 days exudation disappeared in 92% of the patients. In postoperative ovarian cancer patients, a mistletoe preparation statistically increased survival. ... 

The products were tested by the mouse P388 lymphocytic leukemia survival test and by the Ehrlich ascites tumor regression test. The polyphosphazene-plat-inum product showed an inhibition of 86% in the ascites test and a 5/7 survival after the eighth day for the P388 mouse test. 

Antitoxic effect. Sesame oil, adiministered to male Wistar rats, ameliorated hepatic and renal damage in a dose-dependent manner and increased survival in lipopolysaccha-ride-treated rats. It decreased lipid peroxide concentration in serum but not in liver and kidney. Serum nitrite production was unaffected by sesame oil ingestion, and the activity of xanthine oxidase was reduced in the lipopolysaccharide-challenged rats k Anti-tumor activity. Water extract of the dried seed, administered intragastrically to mice at a dose of 50 mg/animal daily for 5 days, was active on CA-Ehrlich-ascites, 18% increase in life-span. Intraperitoneal administration was active on Dalton s lyphoma and CA-Ehrlich-ascites, 19 and 39% increase in life-span, respectively ". Seed oil, administered to rats intraperito-neally with 1,2,5,6-dibenzanthracene or re-tene, was active on sarcoma ". 

Hydrocyanation. Although 2-carbomethoxy-2-cyclopentene-l-one (2) is prone to polymerization, it reacts smoothly with diethylaluminium cyanide to give the adduct 3. This product has the carbon skeleton of sarkomycin (7), an antibiotic active against ascites-type tumors. Selective manipulation of the functionaTgroups resulted in the first regiospecific synthesis of 7. ... 

The progression and growth of ovarian carcinoma are also dependent on chemokine-mediated angiogenesis. In one study, the in vitro production of CXCL8 by five human ovarian cancer lines correlated with tumor neovascularization and cancer-related death when implanted into the peritoneum of immunocompromised mice, whereas VEGF production correlated only with ascites production after implantation, but basic FGF did not correlate with the outcome [70]. This concept was further substantiated in patients with ovarian cancer where ascites fluid angiogenic activity directly correlated to CXCL8 [71]. 

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