scholarly journals Sendai virus causes a rise in intracellular free Ca2+ before cell fusion

1982 ◽  
Vol 206 (3) ◽  
pp. 671-674 ◽  
Author(s):  
Maurice B. Hallett ◽  
Pinhas Fuchs ◽  
Anthony K. Campbell
Keyword(s):  
Cell Fusion ◽  
Human Erythrocyte ◽  
Maximum Rate ◽  
Sendai Virus ◽  
Erythrocyte Ghosts ◽  
Absolute Requirement

1. Sendai virus caused a large increase in the concentration of free Ca2+ within human erythrocyte ghosts detected by the Ca2+-activated photoprotein obelin. 2. The increase in intracellular [Ca2+] preceded fusion. However, fusion could also be observed in the absence of a detectable rise in intracellular free [Ca2+]. 3. It was concluded that the increase in intracellular free [Ca2+] was not an absolute requirement for cell fusion, but may be necessary to produce fusion at the maximum rate.

The role of cell swelling and haemolysis in Sendai virus-induced cell fusion and in the diffusion of incorporated viral antigens

1980 ◽  
Vol 42 (1) ◽  
pp. 153-167
Author(s):  
S. Knutton ◽  
T. Bachi
Keyword(s):  
Cell Fusion ◽  
Erythrocyte Membrane ◽  
Sendai Virus ◽  
Viral Envelope ◽  
Cell Swelling ◽  
Haemolytic Activity ◽  
Erythrocyte Ghosts ◽  
Viral Antigens ◽  
Cell Cell

The role of the haemolytic activity of Sendai virus in cell-cell fusion has been examined in monolayers of human erythrocytes and erythrocyte ghosts fused with either haemolytic or non-haemolytic virus. Morphological observations indicate that cell swelling and haemolysis is a distinct event in cell-cell fusion irrespective of whether it is virally induced or, in the case of non-haemolytic virus, experimentally induced. Osmotic swelling appears to be the driving force by which cells which have established sites of membrane fusion expand such sites to form poly-erythrocytes. Immunofluorescent labelling of viral antigens incorporated into the erythrocyte membrane as a result of viral envelope-cell fusion indicates that diffusion of antigens in the plane of the membrane is restricted in intact erythrocytes and resealed erythrocyte ghosts but not in haemolysed erythrocytes or unsealed ghosts. A perturbation of the erythrocyte membrane resulting from osmotic lysis appears to form a prerequisite for the lateral diffusion of viral elements.

Effect of anionic polymers on fusion of Sendai virus with human erythrocyte ghosts

1992 ◽  
Vol 18 (2) ◽  
pp. 163-177 ◽  
Author(s):  
S. Ohki ◽  
K. Arnold ◽  
N. Srinivasakumar ◽  
T.D. Flanagan
Keyword(s):  
Human Erythrocyte ◽  
Sendai Virus ◽  
Erythrocyte Ghosts ◽  
Anionic Polymers

scholarly journals Quantitative introduction of a given macromolecule into cells by fusion with erythrocyte ghosts using a fluorescence activated cell sorter.

1978 ◽  
Vol 26 (12) ◽  
pp. 1067-1073 ◽  
Author(s):  
E Mekada ◽  
M Yamaizumi ◽  
T Uchida ◽  
Y Okada
Keyword(s):  
Light Scattering ◽  
Bovine Serum Albumin ◽  
Human Erythrocyte ◽  
Mononuclear Cells ◽  
Sendai Virus ◽  
Fluorescence Analysis ◽  
Erythrocyte Ghosts ◽  
Cell Sorter ◽  
Sorting Method ◽  
L Cells

FITC-conjugated bovine serum albumin (FITC-BSA) molecules were quantitatively introduced into human erythrocyte ghosts by gradual hemolysis. When the ghosts and L cells were fused with UV-inactivated HVJ (Sendai virus), FITC-BSA was introduced into the cytoplasm of the L cells and fluorescence could be observed inthe cells with a fluorescence microscope. A mixture of L cells and ghosts was introduced into a fluorescence activated cell sorter (FACS), which could separate the mononuclear cells on the basis of their light-scattering profile. Four distinct populations of mononuclear cells were found by fluorescence analysis. These populations were separated from the cell mixture and found to correspond to cells fused with one, two and three ghosts and unfused cells. After separation, the cells from each population could form colonies in culture. As a given macromolecule can be quantitatively introduced into erythrocyte ghosts with the FITC-BSA, after fusion of these ghosts with cells, this sorting method is useful for separating cells containing a definite number of macromolecules.

Role of Ca2+ in preserving viability of Friend erythroleukaemic cells after ultramicroinjection

1977 ◽  
Vol 27 (1) ◽  
pp. 157-165
Author(s):  
M. Wasserman ◽  
A. Loyter ◽  
R.G. Kulka
Keyword(s):  
Protein Synthesis ◽  
Human Erythrocyte ◽  
Sendai Virus ◽  
Atp Depletion ◽  
Complete Medium ◽  
Erythrocyte Ghosts ◽  
Culture Cells ◽  
Atp Levels ◽  
Ehrlich Ascites Tumour ◽  
High Concentrations

Ca2+ facilitated the fusion by Sendai virus of Friend erythroleukaemic cells and Ehrlich ascites tumour cells but not that of hepatoma tissue culture cells. In the absence of Ca2+ Sendai virus caused the complete depletion of ATP and abolished protein synthesis in Friend erythro-leukaemic cells fused with each other. Addition of high concentrations of Ca2+ (10-20 mM) partially protected the cells from ATP depletion. After a further incubation of cells in complete medium plus 0.2 mM adenine, ATP levels and protein synthesis were restored to 60–85% of those of the untreated control. The protective effect of Ca2+ was used to improve the ultramicroinjection method which involves the fusion of human erythrocyte ghosts with cells. When human erythrocyte ghosts containing high K+ were fused with Friend erythroleukaemic cells in the presence of 10 mM Ca2+ ATP levels and protein synthesis after recovery were about 60–85% of the control. Friend erythroleukaemic cells subjected to ultramicroinjection under these conditions had a cloning efficiency of 75–95% of that of the untreated controls. In these experiments 70–100% of the cells had fused with ghosts. Induction of haemoglobin synthesis by dimethylsulphoxide was unimpaired in cells subjected to ultramicroinjection under the same conditions.

scholarly journals FUSION OF INTACT HUMAN ERYTHROCYTES AND ERYTHROCYTE GHOSTS

1974 ◽  
Vol 63 (1) ◽  
pp. 1-11 ◽  
Author(s):  
Hava Peretz ◽  
Zivia Toister ◽  
Yehudith Laster ◽  
Abraham Loyter
Keyword(s):  
Red Blood Cells ◽  
Clostridium Perfringens ◽  
Human Erythrocyte ◽  
Blood Cells ◽  
Sendai Virus ◽  
Iodoacetic Acid ◽  
Human Erythrocytes ◽  
Erythrocyte Ghosts ◽  
Human Red Blood Cells ◽  
Bivalent Cations

Sendai virus is able to induce the fusion of human erythrocytes. Bivalent cations or ATP are not essential for polyerythrocyte formation. High fusion indices were obtained when Sendai virus was added to cells incubated in the presence of both EDTA and iodoacetic acid. Human erythrocyte ghosts prepared by gradual hemolysis still retain the potential to undergo virus-induced fusion. Fusion of human red blood cells without the addition of viruses was obtained by incubation of erythrocytes at pH 10.5 in the presence of Ca++ (40 mM) or by addition of phospholipase C Clostridium perfringens preparations to cells previously agglutinated or polylysine.

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