Requirement of energy for the cell fusion reaction of ehrlich ascites tumor cells by HVJ

Okada, Yoshio; Murayama, F; Yamada, Kazuko

doi:10.1016/0042-6822(66)90312-6

Cited by 68 publications

(24 citation statements)

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“…At about the same stage of the reaction, fusion of viral envelopes with cell membranes occurs (9); glycoproteins and lipids originating from the envelope are explanted into the cell membranes. Within 30 min of the cellfusion process, the original cell-surface area decreases, the degree of decrease depending on the degree of fusion of the cells; (3). Polykaryocytosis also appears.…”

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“…The cell-fusion phenomenon with hemagglutinating virus of Japan (Sendai virus) (HVJ) (1)(2)(3)(4)(5) seems to offer a suitable experimental system for studies on the biological nature of animal-cell membranes (6). Ehrlich ascites tumor cells rapidly agglutinate on addition of HVJ at 00; this agglutination occurs with adsorption of HVJ virions onto the surface of the cells.…”

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Specific Movement of Cell Membranes Fused with HVJ (Sendai Virus)

Okada

Kim

Maeda

et al. 1974

Proc. Natl. Acad. Sci. U.S.A.

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The cell-fusion phenomenon with hemagglutinating virus of Japan (Sendai virus) (HVJ) (1-5) seems to offer a suitable experimental system for studies on the biological nature of animal-cell membranes (6). Ehrlich ascites tumor cells rapidly agglutinate on addition of HVJ at 00; this agglutination occurs with adsorption of HVJ virions onto the surface of the cells. When cell aggregates are incubated at 370, the viral envelopes react with the cell membranes, the membranes of adjacent cells in cell aggregates fuse together within a few minutes, and spherical fused cells appear within 30 min. The efficiency of cell fusion is a function of the concentration of HVJ added (7).In the course of these reactions, some abnormal conditions for the cells are produced. At an early stage, the cell-membrane structure becomes unstable due to reaction with the virus; decrease in the membrane potential and membrane resistance, with leak of ions through the cell membranes, can be detected electrophysiologically. The instability is reversed with progress of cell fusion (8). At about the same stage of the reaction, fusion of viral envelopes with cell membranes occurs (9); glycoproteins and lipids originating from the envelope are explanted into the cell membranes. Within 30 min of the cellfusion process, the original cell-surface area decreases, the degree of decrease depending on the degree of fusion of the cells; In the present paper, a specific movement of fused cell membranes is deduced from observations on the movement of viral antigens integrated into the membranes of fused cells. MATERIALS AND METHODSAs Ehrlich ascites tumor cells, cells passaged in the abdomen of ddO mice or cells of an established line passaged in vitro were used. Results with the two were similar in the present experiments. The cells were washed and suspended in 9 volumes of balanced salt solution (140 mM NaCl; 54 mM KCl; 0.34 mM Na2HPO4; 0.44 mM KH2PO4; buffered with 10 mM Tris * HCl at pH 7.6) containing 2 mM CaCI2. HVJ, Z strain, was passaged in embryonated eggs, purified by differential centrifugation (2), and then, suspended in balanced salt solution at a concentration of 2000 hemagglutinating units. The virus was inactivated by UV-irradiation before use. A mixture of equal volumes (0.5 ml each) of tumor cells and HVJ was incubated at 00 for 5 min and then the mixture was incubated in a water bath at 370 for 30 min with shaking.After completion of the cell-fusion reaction, cells were washed to remove free virus at 00, and then cultured in minimal essential medium supplemented by 10% calf serum at 37°.When inhibitors were used, they were dissolved in the medium. After culture, cells were washed again and exposed to antiserum against HVJ envelope developed in rabbit at 00 for 20 min. Then the cells were washed at 0°, and mixed with fluorescein isothiocyanate-labeled anti-rabbit IgG 7S antibody (Hyland Biochemicals). After incubation for 20 min at 00, the cells were washed and observed under a microscope with incident-light fluorescence (Leitz). In ...

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confidence: 99%

Specific Movement of Cell Membranes Fused with HVJ (Sendai Virus)

Okada

Kim

Maeda

et al. 1974

Proc. Natl. Acad. Sci. U.S.A.

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“…In a series of experiments, Okada and co-workers introduced methods for quantitating the fusion reaction (fusion index = number of cells prior to the fusion reaction/number of cells after the fusion reaction -1 ; range is generally 0.5 to 12.0), 153 and then proceeded to investigate the effect of various parameters on cell fusion. These experiments demonstrated a requirement for Ca 2 +, 149 dependence on oxygen and oxidative phosphorylation, 151 positive correlation between the amount of virus present and the fusion index, 146 and the variable capacity of different cell lines to respond to the virus with regard to cell fusion. 153 Harris and Watkins first demonstrated the practical application of Sendai virus for the purpose of producing heterokaryons.…”

Section: Formation Of Hybrid Cellsmentioning

confidence: 91%

Linkage Analysis Using Somatic Cell Hybrids

Ruddle

1972

Advances in Human Genetics

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“…Increased activity of enzymes associated with oxidative metabolism has been noted in cell fusion from within, induced by several animal viruses (II). Okada et al (9) reported that ATP and Ca 2 + are essential factors for Sendai virus (HVJ)-induced cell fusion. Polykaryocyte formation in Ehrlich ascites tumor cells was not induced under anaerobic conditions or in the presence of DNP.…”

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confidence: 99%

Requirement of Carbohydrates for Cell Fusion Induced by Vesicular Stomatitis Virus

Takehara

1981

Microbiology and Immunology

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The fusion of BHK-21-KB cells by vesicular stomatitis virus was not induced in Eagle's minimal essential medium without glucose. In medium containing glucose, the rate of polykaryocyte formation decreased as the concentration of glucose was reduced below 5 mM. However, no reduction in virus production 24 hr after infection was seen under this condition. Addition of pyruvate or mannose to the culture medium caused a reversal of cell fusing activity. Cell fusion and virus growth were significantly suppressed by sodium azide and 2,4-dini trophenol.

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Requirement of energy for the cell fusion reaction of ehrlich ascites tumor cells by HVJ

Cited by 68 publications

References 11 publications

Specific Movement of Cell Membranes Fused with HVJ (Sendai Virus)

Specific Movement of Cell Membranes Fused with HVJ (Sendai Virus)

Linkage Analysis Using Somatic Cell Hybrids

Requirement of Carbohydrates for Cell Fusion Induced by Vesicular Stomatitis Virus

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