scholarly journals Engineering a recombinant Herpesvirus saimiri strain by co-culturing transfected and permissive cells

2019 ◽  
pp. 37-44
Author(s):  
A. Hamad ◽  
S.P. Chumakov
Keyword(s):  
Measles Virus ◽  
Selectable Marker ◽  
Human Lymphocytes ◽  
Therapeutic Agents ◽  
Squirrel Monkeys ◽  
Lymphoid Cells ◽  
Herpesvirus Saimiri ◽  
Permissive Cell ◽  
Gamma Herpesvirus ◽  
Packaging Capacity

Recombinant herpesviruses can be used as oncolytic therapeutic agents and high packaging capacity vectors for delivering expression cassettes into the cell. Herpesvirus saimiri is a gamma-herpesvirus that normally infects squirrel monkeys but also has a unique ability to infect and immortalize human lymphocytes while allowing them to retain their mature phenotype and functional activity. Recombination of the Herpesvirus saimiri genome in permissive cells is impeded by its resistance to chemical transfection and electroporation. The aim of this study was to develop an effective method for incorporating expression cassettes into the genome of Herpesvirus saimiri without having to transfect a permissive cell culture. Transfected HEK-293T cells expressing glycoproteins of the measles virus vaccine strain were co-cultured with permissive OMK cells infected with Herpesvirus saimiri. Cell fusion and formation of syncytia stimulated recombination between the viral genome and the expression cassette; this allowed us to obtain a recombinant Herpesvirus saimiri variant without chemical transfection in permissive cells. The genetically modified virus expressed a selectable marker and retained its ability to persist in the cell in the latent state; it also caused immortalization of primary lymphoid cells. The proposed approach allows engineering recombinant Herpesvirus saimiri strains carrying a variety of expression cassettes in its genome.

scholarly journals Measles Virus-Induced Immunosuppression In Vitro Is Independent of Complex Glycosylation of Viral Glycoproteins and of Hemifusion

2000 ◽  
Vol 74 (16) ◽  
pp. 7548-7553 ◽  
Author(s):  
Armin Weidmann ◽  
Christian Fischer ◽  
Shinji Ohgimoto ◽  
Claudia Rüth ◽  
Volker ter Meulen ◽  
...  
Keyword(s):  
Measles Virus ◽  
Human Lymphocytes ◽  
Lymphoid Cells ◽  
Oxygen Intermediates ◽  
Inhibitory Peptides ◽  
Viral Particles ◽  
Infected Cells ◽  
Necessary And Sufficient ◽  
Cellular Fusion

ABSTRACT Expression of the measles virus (MV) F/H complex on the surface of viral particles, infected cells, or cells transfected to express these proteins (presenter cells [PC]) is necessary and sufficient to induce proliferative arrest in both human and rodent lymphoid cells (responder cells [RC]). This inhibition was found to occur independent of apoptosis and soluble mediators excluded by a pore size filter of 200 nm released from either PC or RC. We now show that reactive oxygen intermediates which might be released by RC or PC also do not contribute to MV-induced immunosuppression in vitro. Using an inhibitor of Golgi-resident mannosidases (deoxymannojirimycin), we found that complex glycosylation of the F and H proteins is not required for the induction of proliferative arrest of RC. As revealed by our previous studies, proteolytic cleavage of the MV F protein precursor into its F1 and F2 subunits, but not of F/H-mediated cellular fusion, was found to be required, since fusion-inhibitory peptides such as Z-d-Phe-l-Phe-Gly (Z-fFG) did not interfere with the induction of proliferative inhibition. We now show that Z-fFG inhibits cellular fusion at the stage of hemifusion by preventing lipid mixing of the outer membrane layer. These results provide strong evidence for a receptor-mediated signal elicited by the MV F/H complex which can be uncoupled from its fusogenic activity is required for the induction of proliferative arrest of human lymphocytes.

Oral Excretion of Herpesvirus saimiri In Captive Squirrel Monkeys and Incidence of Infection in Feral Squirrel Monkeys2

1973 ◽  
Vol 51 (6) ◽  
pp. 1987-1989 ◽  
Author(s):  
Lawrence A. Falk ◽  
Stephen Nigida ◽  
Friedrich Deinhardt ◽  
Robert W. Cooper ◽  
Jorge I. Hernandez-Camacho
Keyword(s):  
Squirrel Monkeys ◽  
Herpesvirus Saimiri

Herpesvirus saimiri antigens and virus recovery from cultured cells and antibody levels and virus isolations from squirrel monkeys

1973 ◽  
Vol 38 (2) ◽  
pp. 491-496 ◽  
Author(s):  
Harvey Rabin ◽  
Gary Pearson ◽  
George Klein ◽  
Dharam Ablashi ◽  
William Wallen ◽  
...  
Keyword(s):  
Cultured Cells ◽  
Squirrel Monkeys ◽  
Herpesvirus Saimiri ◽  
Virus Recovery ◽  
Antibody Levels

scholarly journals Generation of biliary lesions after transfer of human lymphocytes into severe combined immunodeficient (SCID) mice.

1989 ◽  
Vol 170 (6) ◽  
pp. 1919-1930 ◽  
Author(s):  
S M Krams ◽  
K Dorshkind ◽  
M E Gershwin
Keyword(s):  
Bile Duct ◽  
Human Lymphocytes ◽  
Severe Combined Immunodeficiency ◽  
Serum Levels ◽  
Scid Mice ◽  
Lymphoid Cells ◽  
Biliary Cirrhosis ◽  
Human Mononuclear Cell ◽  
Mononuclear Infiltrate ◽  
Immunodeficiency Disease

Human PBL have been reported to reconstitute B and T cells as well as human serum Ig in mice with severe combined immunodeficiency disease (SCID). To confirm these observations and attempt the transfer of an autoimmune disease to the immunodeficient animals, groups of SCID mice received an injection of PBL from patients with primary biliary cirrhosis (PBC) or from normal volunteers. By 8 wk after the injection of 10-42 x 10(6) PBL into the mice, human lymphoid cells were detected in the spleen of approximately half of the animals and all had detectable serum levels of human IgG. Moreover, the sera of SCID mice that received cells from patients with PBC contained human antimitochondrial antibodies (AMA) to dihydrolipoamide acetyltransferase, the major mitochondrial autoantigen of PBC. Histologically, a human mononuclear cell infiltrate was present around the portal areas of the liver and inflammation, bile duct atypica, and necrosis of bile duct cells were observed. While the biliary lesions in the SCID recipients of PBC cells were more severe, a mononuclear infiltrate was clearly evident in mice that received cells from normal donors, suggesting the presence of a graft-vs.-host-like disease. While these data are the first to describe an animal model with both the humoral and cellular characteristics of PBC, they also raise an interesting question regarding the preferential localization of lymphoid cells to the biliary system.

scholarly journals Failure to cleave measles virus fusion protein in lymphoid cells. A possible mechanism for viral persistence in lymphocytes

1981 ◽  
Vol 154 (5) ◽  
pp. 1489-1499 ◽  
Author(s):  
RS Fujinami ◽  
MBA Oldstone
Keyword(s):  
Plasma Membrane ◽  
Fusion Protein ◽  
Measles Virus ◽  
Viral Persistence ◽  
Lymphoid Cells ◽  
Lymphoblastoid Cells ◽  
Membrane Enzyme ◽  
Virus Fusion ◽  
Competent Cells ◽  
Daudi Cells

The host-directed cleavage of measles virus fusion protein on infected lymphoid cells was studied to understand the mechanism of viral persistence in lymphoid cells in vivo. Several lymphoblastoid cell lines were infected with measles virus, and the viral glycoproteins expressed on the cell's surface were radiolabeled and analyzed for cleavage of fusion (F(0)) to F(1) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Daudi and Ramos lymphoblastoid cells were deficient in their ability to cleave measles virus fusion protein and correspondingly produced low titers of infectious measles virus, Daudi cells being more defective than Ramos cells. In contrast, other lymphoblastoid cells studied, Victor, Raji, Wi-L2, RPMI 8866, and Seraphine, cleaved the fusion polypeptide and made significantly more infectious virus. Despite their defect in cleaving F protein, Daudi cells were able to assemble and release (noninfectious) measles virus particles into the fluid phase. The deficit in Daudi cells was corrected by fusing infected Daudi cells with cleavage-competent cells such as Victor or Raji. Furthermore, the cleavage event performed by competent cells could be mimicked at the plasma membrane by treating infected Daudi cells with trypsin, implicating the role of a plasma membrane enzyme in cleaving F(0) to F(1) during measles virus infection. Hence, lymphoid cells deficient in the plasma membrane enzyme required to cleave F protein are permissive for measles virus, maintain viral gene products, produce mostly noninfectious virus, and fail to place the biologic activity F(1) protein on their surfaces.

scholarly journals Isolation of a heterogeneous population of temperature-sensitive mutants of measles virus from persistently infected human lymphoblastoid cell lines.

1978 ◽  
Vol 147 (6) ◽  
pp. 1637-1652 ◽  
Author(s):  
G Ju ◽  
S Udem ◽  
B Rager-Zisman ◽  
B R Bloom
Keyword(s):  
Cell Lines ◽  
Measles Virus ◽  
Heterogeneous Population ◽  
Lymphoid Cells ◽  
Temperature Sensitive ◽  
Defective Interfering Particles ◽  
Persistently Infected ◽  
Persistent Infections ◽  
Extracellular Virus ◽  
Temperature Sensitive Mutants

Two human lymphoblastoid B-cell lines, WI-L2 and 8866, were infected with the Edmonston strain of measles virus at a multiplicity of infection of 10(-6), and stable persistent infections were established. By immunofluorescence and electron microscopy, the vast majority of cells from both cell lines were expressing viral antigens and releasing virion-like particles. However, very little infectious virus could be detected at 37 degrees C, either by an infectious centers assay or by titration of supernates from persistently infected cultures. When cultures were shifted to 31 degrees C, the cells released a population of virus that was temperature-sensitive. Clonal analysis of supernatant virus at 31 degrees C revealed a highly heterogeneous population of temperature-sensitive mutants, differing in plating efficiency ratios, thermolability, and antigen production at the nonpermissive temperature. Factors such as interferon, defective interfering particles, and extracellular virus do not appear to be important in maintaining the persistent carrier state. These studies have important implications for persistent infections of lymphoid cells in vivo, and the slow neurological diseases associated with measles, subacute sclerosing panencephalitis, and multiple sclerosis.

scholarly journals Binding, internalization, and hydrolysis of low density lipoprotein in long-term lymphoid cell lines from a normal subject and a patient with homozygous familial hypercholesterolemia.

1976 ◽  
Vol 144 (2) ◽  
pp. 444-455 ◽  
Author(s):  
Y K Ho ◽  
M S Brown ◽  
H J Kayden ◽  
J L Goldstein
Keyword(s):  
Cell Surface ◽  
Familial Hypercholesterolemia ◽  
Human Lymphocytes ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Lymphoid Cells ◽  
Low Density ◽  
High Affinity ◽  
Hydrolysis Of

Long-term established human lymphoid cells were shown to possess high affinity cell surface receptors for low density lipoprotein (LDL), the major cholesterol-carrying protein in human plasma. Binding of LDL to these receptors was followed by internalization of the lipoprotein and hydrolysis of its protein and cholesteryl ester components. Cultured lymphocytes from a patient with the homozygous form of familial hypercholesterolemia lacked cell surface LDL receptors and therefore failed to take up and degrade the lipoprotein with high affinity. Cultured human lymphocytes should prove useful for further studies of: (a) the relation between cholesterol metabolism and cellular function and (b) the mechanism by which LDL binding at the cell surface leads to internalization of the lipoprotein.

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