Search PubMed⌕ Search

Biomedical subjects

O Fodstad

Publications and source records attributed to O Fodstad.

At least 145 records · Page 8Linked to original sources
PubMed1984

Antigen expression in human melanoma cells in relation to growth conditions and cell-cycle distribution.

The expression of a melanoma-associated antigen, recognized by the monoclonal antibody 9.2.27, has been studied in the human FME melanoma cell line, grown as a monolayer under various conditions in vitro and as tumours in athymic mice. Two-parameter flow cytometric measurements of DNA and immunofluorescence showed that the antigen expression was uniform throughout the cell cycle. Highest expression of the melanoma-associated antigen was found in cultures in which the medium was frequently renewed (4 times in 6 days) and no contact inhibition was present. In comparison, the antigen expression of cells subjected to medium starvation (6 days without medium renewal) was reduced to 44%, and the cell size, as measured by forward angle light scatter, to 82% of the values found for optimally growing cells. Intermediate conditions, such as scanty medium supply or contact inhibition of growth, gave smaller effects. Two-parameter measurements of cell size and immunofluorescence demonstrated a positive correlation between cell size and antigen expression. The reduction in cell size, therefore, could explain part, but not all, of the reduction in antigen expression on medium-starved and contact-inhibited cells. The antigen expression on FME cells grown as xenografts in athymic mice varied between individual tumours and was lower than on cells from optimal in vitro growth. Due to smaller cell size, the antigen density was, however, comparable to that on cells from optimal in vitro growth.

Animals↗
PubMed1984

Pharmacological studies of ricin in mice and humans.

A highly sensitive enzyme-linked immunosorbent assay (ELISA) for determination of ricin in serum is presented. Using this method it was found that IV-injected ricin disappeared from the plasma of mice and cancer patients according to first-order kinetics. DBA mice were found to be more sensitive to ricin than C3H and B6D2 mice. When mice of the different strains were given the same dose of ricin, the concentrations found in liver, spleen, and kidneys were highest in the most sensitive mice. Ricin disappeared most rapidly from serum of the mice with the highest sensitivity. The inverse correlation between the rate of disappearance of ricin from serum and the tissue concentrations reached may be due to the fact that ricin is rapidly and firmly bound to cell surface receptors. Whole-body autoradiography after IV injection of 125I-labeled ricin showed the highest amount of radioactivity in liver, spleen, and adrenal cortex. Considerable amounts of radioactivity were also present in bone marrow, showing that the lack of myelosuppressive activity of ricin previously found in mice and dogs cannot be accounted for by the failure of ricin to reach the bone marrow. Part of the ricin in the tissues was present in the form of free chains, the highest fraction being present in the liver. In this tissue both the free A-chains and those present in whole ricin were found to be modified. However, the modified A-chains had retained their full capacity to inhibit protein synthesis in vitro. In cancer patients, toxicity appeared at about the same initial serum levels as in the mice, supporting the view that mouse data have a good predictive value for man. At each dose level the individual variations were modest, a finding that is important for eventual clinical use of this potent drug.

Animals↗
PubMed1984

Immune characteristics of the beige-nude mouse. A model for studying immune surveillance.

Double immunodeficient mice, homozygous for both the beige and the nude genes, were developed on C57BL/6N-beige and N:NIH(S)-nude background through five steps of mating. The animals were healthy and had a life span comparable to that of regular nude mice. Beige and regular nude mice showed a significant difference in natural killer (NK) activity, with mean +/- SE values of 6 +/- 1.0% and 25 +/- 2.8%, respectively. The response to the T-cell mitogens phytohaemagglutinin and concanavalin A was similar in beige and NIH mice of the same genotypes for the nude genes. Both groups of nu/nu mice also showed some response in these assays, as well as in the plaque-forming cell assay, indicating that nude mice are not completely devoid of functional T or T-like cells. The nude genes were found to increase the response to the B-cell mitogen lipopolysaccharide in both the beige and NIH animals, whereas the response was reduced in mice of the beige genotypes. The viable, low NK beige-nude mice here reported on may be a valuable tool for studying factors involved in host defence against tumours. Interestingly, no spontaneous tumours have so far been observed in such mice.

Animals↗
PubMed1984

Lack of correlation between natural killer activity and tumor growth control in nude mice with different immune defects.

To elucidate the in vivo role of natural killer (NK) cells, the growth of several murine and human tumors was studied in four variants of athymic, nude mice with different levels of NK activity. Beige-nude mice, homozygous for both the beige and the nude genes, had very low levels of NK activity, and their response to the B-cell mitogen, bacterial lipopolysaccharide, was lower than that of high-NK, adult NIH nude mice. Young and adult NIH nudes had different NK levels and showed different response in assays for K-cell, T-cell, and B-cell activity. The B-cell-defective NIH-II mice had slightly lower NK levels than adult NIH animals, but much lower response in the antibody-dependent cell-mediated cytotoxicity assay. No correlation was found between host NK activity and the s.c. growth of various human (LOX, CEM, K562) and murine (YAC-1) tumor cells. Low NK activity was not associated with increased lung colony formation in a metastasis model using i.v.-injected human (LOX) and murine (B16F10) melanoma cells. No relationship was found between host NK activity and the rate of elimination of i.v.-injected 5-iodo-2'-deoxyuridine-labeled LOX, B16F10, and YAC-1 cells from lungs, liver, or spleen. The results fail to support the view that NK cells exert significant direct effects on tumor cells in vivo.

Animals↗
PubMed1984

Reduced antineoplastic activity in mice of cisplatin administered with high salt concentration in the vehicle.

The effect of high NaCl concentration in the vehicle on the toxic and antineoplastic activities of cisplatin [cis-dichloro-diammineplatinum(II)] (CDDP) was reinvestigated in mice. The toxicity, as measured by the survival of mice given CDDP iv, was reduced by 50-60% when the NaCl concentration in the vehicle was raised from 0.9 to 4%. In ascitic P388 leukemia the antineoplastic activity of CDDP given ip was not reduced significantly. However, in all other systems studied the antitumor activity was reduced when the CDDP was dissolved in high NaCl solution. The tumor models studied included systemic P388 and L1210 leukemias, Lewis lung carcinoma, and 5 human tumor xenografts. The human tumors were studied by the subrenal capsule assay. In the case of a malignant melanoma and a malignant fibrous histiocytoma, the effect also was demonstrated in the subcutaneous nude mouse model. In one of the malignant melanomas a 50% increase in the CDDP dose did not compensate for the reduced antitumor activity caused by the high NaCl concentration in the vehicle. These results, which stand in contrast to current views, question the experimental basis for the use of high-NaCl vehicles in the "high-dose" CDDP regimens.

Animals↗
PubMed1984

Phase I study of the plant protein ricin.

A Phase I study was carried out with ricin, a plant toxin acting by inhibiting protein synthesis, on 54 cancer patients with advanced disease. Ricin was given as i.v. bolus injections every two weeks at dose levels ranging from 4.5 to 23 micrograms/sq m of estimated body surface area. Ricin was well tolerated at doses up to 18 to 20 micrograms/sq m. At these levels and at higher levels, flu-like symptoms with fatigue and muscular pain appeared and, in some patients, nausea and vomiting occurred also. No myelo-suppression was seen. Antibodies to ricin were detected in serum after two to three ricin injections. Ricin was eliminated from blood according to first order kinetics. At each dose level, the plasma concentrations, as well as the side effects, showed only minor differences between patients. The highest dose given, 23 micrograms/sq m, gave plasma concentrations twice those found previously to be therapeutically effective in tumor-bearing mice. Of 38 evaluable patients, one patient with lymphoma had a partial response. Stable disease was observed in four patients with renal cancers, in two with soft tissue sarcomas, and in one patient each with mesothelioma, thyroid, and rectal cancer. A dose of 23 micrograms/sq m is recommended for Phase II trials of ricin.

Abrin↗
PubMed1983

Antibody formation against the cytotoxic proteins abrin and ricin in humans and mice.

Antibody formation may limit the therapeutic use of cancerostatic proteins. To study the significance of antibody formation against abrin and ricin, highly sensitive ELISA procedures for determination of anti-abrin and anti-ricin were developed. In mice treated weekly with therapeutic doses of ricin, antibodies appeared after 2-3 weeks and then rose rapidly, whereas after abrin treatment the antibody formation was slower. Ricin A-chain was found to be more immunogenic than either intact ricin or human serum albumin (HSA). Cyclophosphamide inhibited the antibody response to both abrin and ricin and a combination of cyclophosphamide and prednisolone totally inhibited both anti-abrin and anti-ricin formation during the 6-week observation period. In mice treated weekly with HSA, abrin treatment strongly reduced the anti-HSA formation, showing that abrin has an immunosuppressive effect which appeared to be stronger than that of cyclophosphamide. The existence of circulating antigen-antibody complexes could be demonstrated in the sera of toxin-treated mice by precipitation with polyethyleneglycol, whenever antibodies were detectable with ELISA. The life-span of animals given lethal ricin doses was appreciably enhanced in animals having antibody levels in excess of 10-20 ng/ml. In cancer patients treated i.v. every second week with therapeutic toxin doses, the 10-20 ng/ml levels of anti-ricin and anti-abrin were reached 6-8 weeks and 7-10 weeks after the first injection of ricin and abrin, respectively. The data indicate that the effective therapeutic use of abrin and ricin as single agents may be limited to these time frames, but that the period of effective use may be substantially prolonged if the toxins are given together with conventional cytostatic agents having immuno-suppressive activity.

Abrin↗
PubMed1982

Colony growth and chemosensitivity in vitro of human melanoma biopsies. Relationship to clinical parameters.

To study the usefulness of an in vitro colony-forming assay in predicting individual clinical responses to chemotherapy, tumor cells obtained from 150 melanoma metastases (119 patients) were grown in soft agar according to the method of Courtenay and Mills (1978), and tested for sensitivity to DTIC, CCNU, vinblastine, procarbazine, abrin and ricin. In 83% of the cases colony formation was observed (plating efficiency, PE, greater than 0.01%). Twenty-seven per cent of the tumors gave PEs greater than 1%, 45% gave PEs in the range 0.1-0.9%, whereas 11% of the tumors gave 0.01-0.09%. The PEs were not correlated with the degree of pigmentation or with the clinical course. Evaluable chemosensitivity data were obtained on 104 metastases from 83 patients. Large differences in sensitivity were seen. In cases which were evaluable both in vivo and in vitro a clear correlation was found between the in vitro chemosensitivity, expressed as the expected growth delay, and the clinical response to chemotherapy. Tumors from patients with partial response, mixed response or stable disease after prior progression, all had rather high in vitro sensitivity to the drug used (expected growth delay greater than 2.0), whereas patients with progression had lower sensitivity. The results confirm that the soft agar method used here provides good culture conditions for human melanoma cells and show that chemosensitivity data can be obtained in a high percentage of melanoma patients. The approach used seems promising in aiding clinicians to adjust chemotherapy to individual patients.

Antineoplastic Agents↗
PubMed1982

Growth characteristics of human melanoma xenografts.

The growth of twelve human malignant melanomas in athymic nude mice was studied. Gompertz curves were fitted to volumetric growth data. DNA histograms were obtained with flow cytometry. Each of the twelve melanomas exhibited a characteristic growth pattern, indicating that inherent properties of the tumours are important for the growth control. The theoretical maximum volumes (Vmax) ranged from 208 to 12,900 mm3, the volume doubling times (Td) from 2.8 to 15.3 days (V = 50 mm3) and from 3.8 to 64.6 days (V = 200 mm3), and the fraction of cells in S from 5 to 21%. Tumours with short Td were characterized by a higher growth fraction and probably by a lower cell loss factor than those with long Td. The growth was also influenced by the nude mouse host, as indicated by the values for Vmax, which were similar to those reported for mouse tumours (geometric mean = 8100 mm3), but considerably lower than the volumes of many tumours in man. Also the Td-values for the xenografts were generally lower than those reported for tumours in man, presumably due to a lower cell loss factor. During serial transplantation the growth rate of one of the melanomas increased abruptly, probably because of both an increased growth fraction and a reduced cell loss factor. The latter result demonstrates the necessity of keeping basic biological parameters of xenografts under observation during serial transplantation.

Animals↗
PubMed1981

The usefulness of human tumor cell lines in the study of chemosensitivity. A study of malignant melanomas.

The chemosensitivity of human melanoma cells has been studied before and after continuous in vitro culture. Altogether, nine cell lines were studied, two derived from patients' biopsies, and seven from xenografts in athymic mice. The sensitivity to the agents DTIC (Dacarbazine), CCNU (Lomustine), procarbazine, vinblastine, abrin and ricin was assayed. Furthermore, in five cases the chemosensitivity of the cell lines was compared to that of tumors obtained by injecting the cell lines into athymic mice. In all cases the sensitivity was measured in an in vitro soft agar assay. Upon cultivation in vitro, two of the cell lines, one derived from a patient's metastasis and one from a xenograft in athymic mice, showed marked increases in sensitivity to some of the drugs, whereas sensitivity to other drugs showed little or no change. For the other cell lines small, but definite increases or decreases in chemosensitivity were observed. Permanent cultures showed the same chemosensitivity as early subcultures. The tumors formed by injecting the cell lines into athymic mice showed moderate changes in chemosensitivity, as compared to the cell lines in vitro. The data indicate that considerable changes in chemosensitivity may occur when cells are brought from in vitro to in vitro conditions and vice versa and that such changes may be highly specific. Therefore, although cell lines may be useful in some respects, they should be used with caution in attempts to evaluate quantitatively the sensitivity of human tumors to cancerostatic drugs.

Animals↗
PubMed1981

Cultivation of human melanomas in soft agar. Factors influencing plating efficiency and chemosensitivity.

As part of a programme to study the predictive clinical value of a soft agar assay for measuring chemosensitivity of human melanomas in vitro, we have observed the effect of three disaggregation methods on the yield of tumor cells, plating efficiency in soft agar and chemosensitivity. The yields and plating efficiencies obtained, as well as sensitivity to DTIC, CCNU, vinblastine and abrin, were about the same whether collagenase/pronase/DNase-treatment, trypsin/EDTA-treatment or mechanical treatment was used. When melanoma xenografts of different sizes were studied, an inverse relationship between tumor size and plating efficiency was found, whereas chemosensitivity was unaffected by tumor size. The highest plating efficiencies of melanoma cells, both from patient biopsies and from xenografts, were obtained when red blood cells were present and a low oxygen concentration (5%) was used. The results demonstrate that, in the case of melanomas, the fraction of tumor cells that are clonogenic in vitro depends on the size of the tumors, and even more so on the culture conditions used. An important finding was that chemosensitivity in vitro appears to be unaffected by the disaggregation method and by tumor size.

Abrin↗
PubMed1981

Comparison of two soft-agar methods for assaying chemosensitivity of human tumours in vitro: malignant melanomas.

Two soft-agar methods for assaying chemosensitivity of human cancers in vitro were compared with respect to colony morphology, plating efficiency (PE) and chemosensitivity of human melanomas. In 9 xenografts and 9 patients' biopsy specimens Method A (essentially that of Courtenay & Mills, 1978) gave considerably higher PE that Method B (essentially that of Hamburger & Salmon, 1977) and, in contrast to Method B, the number of colonies was proportional to the number of cells plated. Evidence was obtained that the observed differences in PE could be attributed to the low O2 concentration and the presence of rat red blood cells in Method A. Colony morphology was similar in the 2 assays. When cells from 4 xenografted melanomas were treated in vitro with DTIC, CCNU, vinblastine and abrin, and the inhibition of colony formation was assayed concurrently in the 2 soft-agar methods, the tumour cells appeared to be more sensitive to 3 of the drugs in Method B than in A. The results demonstrate that chemosensitivity data obtained with the 2 assays cannot be directly compared.

Abrin↗
PubMed1980

In vitro sensitivity of human melanoma xenografts to cytotoxic drugs. Correlation with in vivo chemosensitivity.

Single-cell suspensions prepared from five human melanomas, grown serially as xenografts in athymic nude mice, were exposed in vitro to increasing concentrations of DTIC (Dacarbazine), CCNU (Lomustine), procarbazine, vinblastine, and the cancerostatic lectins abrin and ricin. The in vitro chemosensitivity of the cells, as measured by the drug concentrations required to inhibit colony formation in soft agar by 50%, was correlated with the growth delay of the xenografts in vivo, previously observed after treatment of the animals with maximum tolerable doses of the same drugs. It was found that for each drug the in vitro sensitivity of the different xenografts was strongly correlated with their response in vivo. The results provide evidence that the soft agar test, as carried out here, adequately reflects the relative sensitivity of the xenografts in vivo. The data indicate that human xenografts may be used to develop quantitative in vitro chemosensitivity tests.

Abrin↗
PubMed1980

Response to chemotherapy of human, malignant melanoma xenografts in athymic, nude mice.

In attempts to elucidate the factors determining individual differences in response of tumors to chemotherapeutic agents, the sensitivity of six human malignant melanomas growing in athymic, nude mice was studied. Six agents, viz. DTIC, CCNU, vinblastine, procarbazine, as well as the toxic lectins abrin and ricin, were administered in maximum tolerable doses to the tumor-bearing mice. The response to treatment was expressed as tumor growth delay, i.e. the number of volume double times saved by the treatment. The xenografts had very nearly retined the morphology of the parent tumors and were histologically similar. They showed different and characteristic early growth rates and also wide variations in their response to the agents tested. Unexpectedly, in the case of DTIC, CCNU and procarbazine, the response of the xenografts proved to be inversely related to the early growth rates of the tumors. For vinblastine, abrin and ricin, no correlation between response and growth rate of the tumors was apparent. Procarbazine had the best overall effect among the agents here tested. DTIC, CCNU and vinblastine had somewhat less and about equal overall efficiency. The plant toxin abrin, which acts by inhibiting protein synthesis, was at least as effective as DTIC. When abrin was given together with DTIC, the effect on the two xenografts tested was superior to that of each agent given alone. The wide variations observed in the response of the histologically similar xenografts to the different agents demonstrate that testing of the chemosensitivity of human xenografts requires the use of a panel of tumors of each histological tpye. The clear relationship found between the early growth rate of the xenografts and their sensitivity to three of the drugs most commonly used in the treatment of malignant melanomas, may have clinical implications.

Abrin↗
PubMed1980

A human melanoma cell line established from xenograft in athymic mice.

A human melanoma cell line with unusually high growth potential was established from a xenograft growing in athymic mice. When xenograft fragments were cultured in vitro, melanoma cells grew out rapidly without any contamination of mouse stromal cells. An established cell line, FME, derived from this tumour, grew both in monolayer and in shaker suspension culture with doubling times of about 20 h. The cells grew easily at low serum concentrations and could even be cultured in serum-free medium supplemented with insulin and transferrin. The cultured cells were hyperdiploid, as were the cells of the xenograft. The cells grew easily in soft agar and formed tumours in athymic mice. When growing exponentially, the cells were almost unpigmented, but when grown to high density, their melanin content increased. Upon treatment with dimethyl sulphoxide (DMSO), retinoic acid and theophylline, as well as with the tumour promoter 12-O-tetradecanoyl phorbol-13-acetate (TPA), the cells showed growth inhibition and increased melanin synthesis.

Animals↗