Scanning electron microscopy of antibody-dependent cell-mediated cytotoxicity of Friend leukemia virus-infected spleen cells

The structure of Friend leukemia virus (FLV) has been studied by electron microscopy. The virus was harvested from the plasma of infected mice and purified by techniques including differential velocity and isopycnic sucrose gradient centrifugation. Viral nucleic acid was isolated using GuHCl followed by precipitation with ethanol and resuspension in 1 mM dithiothreitol. In initial experiments, the RNA solution was diluted 1:10 into either H2O or 4-8 M urea. Prior to spreading, 1 M ammonium acetate and cytochrome c were added to a final concentration of 0.4 M and 0.01% respectively. Fifty μ1 of this mixture was spread using a glass slide inclined in a ten cm2 trough filled with 15 mM ammonium acetate. Samples were picked up on collodion filmed grids, stained with uranyl acetate and rotary shadowed with Pt:Pd. This method is very similar to the conditions used to study the RNA of avian myeloblastosis virus, murine leukemia virus, and mammary tumor virus (MTV).

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Cell-free transmission of Fv-4 resistance gene product controlling Friend leukemia virus-induced leukemogenesis: a unique mechanism for interference with viral infection

Blood ◽

10.1182/blood.v86.4.1557.bloodjournal8641557 ◽

1995 ◽

Vol 86 (4) ◽

pp. 1557-1563 ◽

Cited By ~ 14

Author(s):

M Kitagawa ◽

S Aizawa ◽

H Kamisaku ◽

H Ikeda ◽

K Hirokawa ◽

...

Keyword(s):

Bone Marrow ◽

Mononuclear Cells ◽

Specific Binding ◽

Leukemia Virus ◽

Spleen Cells ◽

Peripheral Blood Mononuclear ◽

Friend Leukemia ◽

Bone Marrow Chimeras ◽

Friend Leukemia Virus

Fv-4 is a mouse gene that dominantly confers resistance to infection by ecotropic murine leukemia virus (MuLV). We previously demonstrated that mixed radiation bone marrow chimeras containing Fv-4r-bearing BALB/c-Fv- 4Wr (C4W) bone marrow and Fv-4r-bearing C3H/He (C3H) bone marrow grafted into C3H recipient mice (C4W+C3H-->C3H) were resistant to Friend leukemia virus (FLV)-induced leukemogenesis, even when they contained as high as 70% C3H-derived cells. This indicates that FLV- sensitive C3H-derived cells are rendered refractory to infection and/or transformation with FLV when they coexist in mice with Fv-4r-bearing cells. To investigate the mechanism of Fv-4 resistance to FLV-induced leukemogenesis, we first examined the expression of Fv-4r env antigen in the peripheral blood mononuclear cells (PBMC) of these chimeras. The Fv-4r env antigen was present not only on C4W-derived cells, but also on Fv-4r-bearing C3H-derived cells in C4W+C3H-->C3H mixed bone marrow chimeras. The Fv-4r env antigen that binds to the cells surface of C3H cells was found in sera from normal C4W mice, C4W-->C3H chimeras, and C4W+C3H-->C3H mixed chimeras. The serum Fv-4r env antigen binds to ecotropic MuLV receptors, shown by specific binding to transfectant mink cells expressing ecotropic MuLV receptor, but not to parental mink cells. To determine whether the binding of Fv-4r env antigen to the putative MuLV receptors would block FLV infection, C3H thymocytes or spleen cells that had been preincubated with C4W serum were mixed with FLV and the subsequent production of MuLV specific antigens was examined. C3H thymocytes or spleen cells treated with C4W serum became refractory to binding by FLV. These results provide evidence that the Fv-4r env antigen is released from C4W-derived cells in vivo and binds to cells expressing surface receptors for ecotropic MuLV, thereby protecting them from infection with FLV. The implication of these findings for gene therapy of retrovirus-induced disease such as acquired immune deficiency syndrome (AIDS) is discussed.

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Erythroleukemia induction by Friend leukemia virus. A host gene locus controlling early anemia or polycythemia and the rate of proliferation of late erythroid cells.

Journal of Experimental Medicine ◽

10.1084/jem.156.2.398 ◽

1982 ◽

Vol 156 (2) ◽

pp. 398-414 ◽

Cited By ~ 10

Author(s):

T Shibuya ◽

Y Niho ◽

T W Mak

Keyword(s):

Leukemia Virus ◽

Erythroid Cell ◽

Direct Result ◽

Erythroid Cells ◽

Spleen Cells ◽

Erythroid Progenitor ◽

Erythroid Precursor ◽

Cba Mice ◽

Friend Leukemia ◽

Friend Leukemia Virus

This report confirms that the Fv-5 locus controls the types of erythropoiesis induced by Friend erythroleukemia virus (FLV) (21) and extends the study to investigate the mode of action of this locus. With the use of FLV obtained by a variety of procedures, we showed that the polycythemia spleen focus-forming component (SFFVp) was responsible for the contrasting changes of hematocrits observed in FV-Pp (polycythemia strain)-infected DBA/2 (Fv-5pp) or CBA (Fv-5aa) mice. These changes in hematocrits were found to be a direct result of the rise in circulating reticulocytes and erythrocytes in DBA/2 mice and a corresponding drop of these erythroid cells in CBA mice 2 wk after infection. Examination of the FV-P-induced cellular changes indicated that dramatic increase in erythropoietin (epo)-independent erythroid precursor (CFU-E*) cells was detected in the spleens and marrow of both strains of mice. The epo responsiveness of the CFU-E in the uninfected and FV-P-infected CBA and DBA/2 mice was also very similar. Similar to FLV-infected DBA/2 mice, the FV-P-infected CBA mice also developed tumorogenic cells (CFU-FV) relatively early after infection (4-6 wk). Study of the physiological and pathological changes in the marrows and spleens of these infected mice indicated that significant differences were found in the spleens of the two strains of mice. The percent of reticulocytes in the spleen cells of CBA mice remained between 10 and 20%, and level of the DBA/2 mice increased to approximately 50%. This higher rate of erythropoiesis was also reflected in the significantly higher rate of uptake of 59Fe in the spleens of the DBA/2 mice. These results suggest that the Fv-5 locus might control the hematocrit levels of these mice by regulating the rates of erythropoiesis in the spleen levels of these mice, probably by affecting the rate of proliferation of an erythroid cell or cells. The erythroid cell(s) affected is likely to be more mature than the erythroid progenitor, CFU-E, as the levels of CFU-E in these two strains of mice were similar. The hypothesis that Fv-5 may control the rates of proliferation of a late erythroid (cell(s) is also supported by the significantly higher spleen weights found in the infected DBA/2 (approximately 2.5 g/spleen) mice than in the CBA (approximately 1 g/spleen) strain.

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Pathogenesis of Human Hair Defects

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010005007x ◽

1974 ◽

Vol 32 ◽

pp. 12-13

Author(s):

P.S. Porter ◽

T. Aoyagi ◽

R. Matta

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Human Hair ◽

Standard Techniques ◽

Scanning Electron

Using standard techniques of scanning electron microscopy (SEM), over 1000 human hair defects have been studied. In several of the defects, the pathogenesis of the abnormality has been clarified using these techniques. It is the purpose of this paper to present several distinct morphologic abnormalities of hair and to discuss their pathogenesis as elucidated through techniques of scanning electron microscopy.

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Ultrastructural Analysis of the Salivary Glands of Gromphadorhina Portentosa (Schaum)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100080614 ◽

1977 ◽

Vol 35 ◽

pp. 638-639

Author(s):

P.J. Dailey

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Salivary Glands ◽

Secretory Vesicles ◽

The Past ◽

Transmission Electron ◽

Means Of Transmission ◽

Gromphadorhina Portentosa ◽

Scanning Electron ◽

Topographical Relief

The structure of insect salivary glands has been extensively investigated during the past decade; however, none have attempted scanning electron microscopy (SEM) in ultrastructural examinations of these secretory organs. This study correlates fine structure by means of SEM cryofractography with that of thin-sectioned epoxy embedded material observed by means of transmission electron microscopy (TEM).Salivary glands of Gromphadorhina portentosa were excised and immediately submerged in cold (4°C) paraformaldehyde-glutaraldehyde fixative1 for 2 hr, washed and post-fixed in 1 per cent 0s04 in phosphosphate buffer (4°C for 2 hr). After ethanolic dehydration half of the samples were embedded in Epon 812 for TEM and half cryofractured and subsequently critical point dried for SEM. Dried specimens were mounted on aluminum stubs and coated with approximately 150 Å of gold in a cold sputtering apparatus.Figure 1 shows a cryofractured plane through a salivary acinus revealing topographical relief of secretory vesicles.

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