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Cytokine production by human malignant fibrous histiocytoma cell lines
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023-(21Tumor biology
7. Augmentation of Vaccination Effects of PGE2-producing Tumor Cells by Transfection with Cytokine Genes
Ohiro, ](..1,2, Kuramitsu, y2, Kobayashi, M), Hosokawa, 311.z, Totsuka, Y) 1Second Department of Oral Surgery, Hokkaido University School of Dentistry, 2Laboratory of Pathology, Cancer Institute, Hokkaido University School of Medicine, Japan To determine whether cytokines could produce vaccination effects by reversing the suppressed immunogenicity of QRpP cells of a murine fibrosarcoma, which secrete high levels of PGE2, we transfected the IL-2, !FN-g and TNF-a gene respectively, into QRpP ceils. After selection in G-418-containing medium, we isolated clonal cells by the limiting dilution method, mRNAs expression of the transfected cytokine genes was assessed using RT-PCR, and the secretion of cytokines and PGE2 into the supernatant was determined using bioassay and enzyme-linked immnnosorbent assay, respectively. In this way, clones producing high levels of cytokines were selegted. These clones secreted PGE2 into the supernatant just as much as unmodified parental tumor cells. However, all of the clonal cells had lowered tumorigenicity. The mice which rejected an implantation of QRpP-IL-2 clohal cells also rejected an implantation of the parental tumor cells. The activity of pre-cytotoxic T cells (pre-CTLs) obtained from the mice implanted with QRpP-IL-2 clone cells was higher than that of the mice implanted with the other transfectants. These results suggest that transfection with the gene for IL-2 may be an effective strategy for producing a vaccination effect against tumors that secrete immunosuppressive factors such as PGE2.
8. Proliferation and Viability of Malignant and Normal Human Cells Under Hyperthermic Culture Conditions
lwagami, yl, Samukawa, 1.1, Wada, 1.1, Miyata, 1(.i, Morita, N. I, Sakamoto, 1.1, Okumura, 1t. 2 1Department of Dentistry and Oral Surgery, Wakayama Medical College, 2Second Department of Surgery, School of Medicine, Nihon University, Japan The idea of cancer hyperthermia was developed on the basis of the property of cancer cells being more sensitive to high temperatures than normal cells. However, the nature of this property of cancer cells has not been adequately elucidated. Moreover, a phenomenon in which the response to a hightemperature environment varies with the type of cell has also been reported, but this phenomenon also remains insufficiently explained. We therefore conducted experiments on two normal human diploid cell strains and three malignant human cell lines. The experiments centered on observations of changes in cell proliferation and viability in addition to changes in other biological properties. The results can be summarized as follows: 1) All of the
malignant human cell lines were found to be more sensitive to hyperthermia than the normal human diploid cell strains. 2) Slight differences in proliferation were observed according to the malignant cell line, with the adenocarcinoma cell line being the most sensitive. 3) There was a correlation between the changes in proliferation and viability. We discuss and report on the sensitivity of human cells to hyperthermia based on these findings.
9. Cytokine Production by Human Malignant Fibrous Histioeytoma Cell Lines
Mori, A., Inui, M., Sekita, M., Goto, A., Kageyama, 1., Higuchi, Y, Tagawa, 11. Department of Oral and Maxillofacial Surgery, Faculty of Medicine, Mie University, Mie, Japan Malignant fibrous histiocytoma (MFH) is mainly composed of fibroblastic cells and histiocytic cells, with a variable number of inflammatory cells often being present. It has been demonstrated that some MFH cells may produce cytokines and thus modify the clinicopathological features of this tumor. We examined the production of cytokines in cell lines established from a human maxillary MFH. Materials and Methods: Four cell lines (GN1, GN3, GNCL-1, and GN-CL-2) derived from a serially transplantable M F H tumor (MFH-R) in nude mice and primarily cultured cells (PC) from M F H - R were used. Cells were maintained in DMEM/Ham's F12 (1 : 1) medium containing 10% FBS. The levels of various cytokines, including granulocyte colonystimulating factor (G-CSF), tumor necrosis factor (TNF)a, interleukin (IL)-la, IL-6, platelet-derived growth factor (PDGF)-AB, and transforming growth factor (TGF)-cq were measured in 48-h culture supernatants by enzyme-linked immunosorbent assay. Results: G-CSE IL-la, and IL-6 were detected in all cultures except GN-CL-2, and G-CSF levels in GN1 and GNCL-1 cultures were >50,000 pg/ml. TNF-a was detected in GN1, GN-CL-1, and PC cultures. PDGF-AB was detected in GN3 and GN-CL-2 cultures. TGF-c~ was not detected at all. Conclusion: We confirmed the production of several cytokines by MFH cells. In particular, G-CSF was produced by 4/5 cell lines and showed high levels in GN1 and GN-CL-1 cultures. Leukocytosis was observed in nude mice with MFH-R, GN1, and GN-CL-1 tumors transplanted, but not in mice with GN-CL-2 tumors, and was considered to be due to G-CSF release by M F H cells.
023-(21Tumor biology
7. Augmentation of Vaccination Effects of PGE2-producing Tumor Cells by Transfection with Cytokine Genes
Ohiro, ](..1,2, Kuramitsu, y2, Kobayashi, M), Hosokawa, 311.z, Totsuka, Y) 1Second Department of Oral Surgery, Hokkaido University School of Dentistry, 2Laboratory of Pathology, Cancer Institute, Hokkaido University School of Medicine, Japan To determine whether cytokines could produce vaccination effects by reversing the suppressed immunogenicity of QRpP cells of a murine fibrosarcoma, which secrete high levels of PGE2, we transfected the IL-2, !FN-g and TNF-a gene respectively, into QRpP ceils. After selection in G-418-containing medium, we isolated clonal cells by the limiting dilution method, mRNAs expression of the transfected cytokine genes was assessed using RT-PCR, and the secretion of cytokines and PGE2 into the supernatant was determined using bioassay and enzyme-linked immnnosorbent assay, respectively. In this way, clones producing high levels of cytokines were selegted. These clones secreted PGE2 into the supernatant just as much as unmodified parental tumor cells. However, all of the clonal cells had lowered tumorigenicity. The mice which rejected an implantation of QRpP-IL-2 clohal cells also rejected an implantation of the parental tumor cells. The activity of pre-cytotoxic T cells (pre-CTLs) obtained from the mice implanted with QRpP-IL-2 clone cells was higher than that of the mice implanted with the other transfectants. These results suggest that transfection with the gene for IL-2 may be an effective strategy for producing a vaccination effect against tumors that secrete immunosuppressive factors such as PGE2.
8. Proliferation and Viability of Malignant and Normal Human Cells Under Hyperthermic Culture Conditions
lwagami, yl, Samukawa, 1.1, Wada, 1.1, Miyata, 1(.i, Morita, N. I, Sakamoto, 1.1, Okumura, 1t. 2 1Department of Dentistry and Oral Surgery, Wakayama Medical College, 2Second Department of Surgery, School of Medicine, Nihon University, Japan The idea of cancer hyperthermia was developed on the basis of the property of cancer cells being more sensitive to high temperatures than normal cells. However, the nature of this property of cancer cells has not been adequately elucidated. Moreover, a phenomenon in which the response to a hightemperature environment varies with the type of cell has also been reported, but this phenomenon also remains insufficiently explained. We therefore conducted experiments on two normal human diploid cell strains and three malignant human cell lines. The experiments centered on observations of changes in cell proliferation and viability in addition to changes in other biological properties. The results can be summarized as follows: 1) All of the
malignant human cell lines were found to be more sensitive to hyperthermia than the normal human diploid cell strains. 2) Slight differences in proliferation were observed according to the malignant cell line, with the adenocarcinoma cell line being the most sensitive. 3) There was a correlation between the changes in proliferation and viability. We discuss and report on the sensitivity of human cells to hyperthermia based on these findings.
9. Cytokine Production by Human Malignant Fibrous Histioeytoma Cell Lines
Mori, A., Inui, M., Sekita, M., Goto, A., Kageyama, 1., Higuchi, Y, Tagawa, 11. Department of Oral and Maxillofacial Surgery, Faculty of Medicine, Mie University, Mie, Japan Malignant fibrous histiocytoma (MFH) is mainly composed of fibroblastic cells and histiocytic cells, with a variable number of inflammatory cells often being present. It has been demonstrated that some MFH cells may produce cytokines and thus modify the clinicopathological features of this tumor. We examined the production of cytokines in cell lines established from a human maxillary MFH. Materials and Methods: Four cell lines (GN1, GN3, GNCL-1, and GN-CL-2) derived from a serially transplantable M F H tumor (MFH-R) in nude mice and primarily cultured cells (PC) from M F H - R were used. Cells were maintained in DMEM/Ham's F12 (1 : 1) medium containing 10% FBS. The levels of various cytokines, including granulocyte colonystimulating factor (G-CSF), tumor necrosis factor (TNF)a, interleukin (IL)-la, IL-6, platelet-derived growth factor (PDGF)-AB, and transforming growth factor (TGF)-cq were measured in 48-h culture supernatants by enzyme-linked immunosorbent assay. Results: G-CSE IL-la, and IL-6 were detected in all cultures except GN-CL-2, and G-CSF levels in GN1 and GNCL-1 cultures were >50,000 pg/ml. TNF-a was detected in GN1, GN-CL-1, and PC cultures. PDGF-AB was detected in GN3 and GN-CL-2 cultures. TGF-c~ was not detected at all. Conclusion: We confirmed the production of several cytokines by MFH cells. In particular, G-CSF was produced by 4/5 cell lines and showed high levels in GN1 and GN-CL-1 cultures. Leukocytosis was observed in nude mice with MFH-R, GN1, and GN-CL-1 tumors transplanted, but not in mice with GN-CL-2 tumors, and was considered to be due to G-CSF release by M F H cells.