Biological Properties of H5 Hemagglutinin Expressed by Vaccinia Virus Vector and its Immunological Reactivity with Human Sera

2013 ◽  
Vol 26 (1) ◽  
pp. 49-59 ◽  
Author(s):  
Pirom Noisumdaeng ◽  
Phisanu Pooruk ◽  
Alita Kongchanagul ◽  
Susan Assanasen ◽  
Rungrueng Kitphati ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Biological Properties ◽  
Virus Vector ◽  
Immunological Reactivity ◽  
Human Sera ◽  
Vaccinia Virus Vector

scholarly journals Processing and secretion of nerve growth factor: expression in mammalian cells with a vaccinia virus vector.

1988 ◽  
Vol 8 (6) ◽  
pp. 2456-2464 ◽  
Author(s):  
R H Edwards ◽  
M J Selby ◽  
W C Mobley ◽  
S L Weinrich ◽  
D E Hruby ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cyclic Amp ◽  
Vaccinia Virus ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Signal Peptide ◽  
Virus Vector ◽  
Mammalian Cell Lines ◽  
Vaccinia Virus Vector

To study posttranslational mechanisms for the control of nerve growth factor (NGF), we used a recombinant vaccinia virus vector to independently express the two major NGF transcripts in a variety of mammalian cell lines. The two major transcripts contain NGF (12.5 kilodaltons [kDa]) at the C-terminus and differ by alternative splicing of an N-terminal exon, so that the large precursor (34 kDa) had 67 amino acids upstream of an internal signal peptide and the smaller precursor (27 kDa) had this signal peptide at its N-terminus. In L929 cells, expression of either NGF transcript with the vaccinia virus vector gave rise to an apparently identical intracellular 35-kDa glycosylated precursor formed by cleavage of the primary gene product after the signal peptide. These cells also secreted biologically active NGF. To determine whether NGF processing is restricted by cell type, we infected a variety of mammalian cell lines with both recombinant viruses; all accumulated the same 35-kDa precursor and secreted NGF. Thus, many types of cells have the machinery to process and secrete NGF. However, NGF accumulated intracellularly (presumably in secretory granules) in cells with a regulated pathway of secretion (e.g., AtT-20 and HIT cells). In these cells, a membrane-permeable cyclic AMP analog, 8-bromo-cyclic AMP, stimulated NGF secretion. This suggests a mechanism for the regulation of NGF levels in which specific secretagogues, e.g., neurotransmitters, control NGF secretion.

scholarly journals Potent and Broadly Reactive HIV-2 Neutralizing Antibodies Elicited by a Vaccinia Virus Vector Prime-C2V3C3 Polypeptide Boost Immunization Strategy

2010 ◽  
Vol 84 (23) ◽  
pp. 12429-12436 ◽  
Author(s):  
José Maria Marcelino ◽  
Pedro Borrego ◽  
Cheila Rocha ◽  
Helena Barroso ◽  
Alexandre Quintas ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Neutralizing Antibodies ◽  
Virus Vector ◽  
Recombinant Polypeptide ◽  
Immunization Strategy ◽  
Loop Sequence ◽  
Immunodeficiency Virus ◽  
Boost Immunization ◽  
Vaccinia Virus Vector

ABSTRACT Human immunodeficiency virus type 2 (HIV-2) infection affects about 1 to 2 million individuals, the majority living in West Africa, Europe, and India. As for HIV-1, new strategies for the prevention of HIV-2 infection are needed. Our aim was to produce new vaccine immunogens that elicit the production of broadly reactive HIV-2 neutralizing antibodies (NAbs). Native and truncated envelope proteins from the reference HIV-2ALI isolate were expressed in vaccinia virus or in bacteria. This source isolate was used due to its unique phenotype combining CD4 independence and CCR5 usage. NAbs were not elicited in BALB/c mice by single immunization with a truncated and fully glycosylated envelope gp125 (gp125t) or a recombinant polypeptide comprising the C2, V3, and C3 envelope regions (rpC2-C3). A strong and broad NAb response was, however, elicited in mice primed with gp125t expressed in vaccinia virus and boosted with rpC2-C3. Serum from these animals potently neutralized (median 50% neutralizing titer, 3,200) six of six highly divergent primary HIV-2 isolates. Coreceptor usage and the V3 sequence of NAb-sensitive isolates were similar to that of the vaccinating immunogen (HIV-2ALI). In contrast, NAbs were not reactive on three X4 isolates that displayed major changes in V3 loop sequence and structure. Collectively, our findings demonstrate that broadly reactive HIV-2 NAbs can be elicited by using a vaccinia virus vector-prime/rpC2-C3-boost immunization strategy and suggest a potential relationship between escape to neutralization and cell tropism.

Processing and secretion of nerve growth factor: expression in mammalian cells with a vaccinia virus vector

1988 ◽  
Vol 8 (6) ◽  
pp. 2456-2464
Author(s):  
R H Edwards ◽  
M J Selby ◽  
W C Mobley ◽  
S L Weinrich ◽  
D E Hruby ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cyclic Amp ◽  
Vaccinia Virus ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Signal Peptide ◽  
Virus Vector ◽  
Mammalian Cell Lines ◽  
Vaccinia Virus Vector

To study posttranslational mechanisms for the control of nerve growth factor (NGF), we used a recombinant vaccinia virus vector to independently express the two major NGF transcripts in a variety of mammalian cell lines. The two major transcripts contain NGF (12.5 kilodaltons [kDa]) at the C-terminus and differ by alternative splicing of an N-terminal exon, so that the large precursor (34 kDa) had 67 amino acids upstream of an internal signal peptide and the smaller precursor (27 kDa) had this signal peptide at its N-terminus. In L929 cells, expression of either NGF transcript with the vaccinia virus vector gave rise to an apparently identical intracellular 35-kDa glycosylated precursor formed by cleavage of the primary gene product after the signal peptide. These cells also secreted biologically active NGF. To determine whether NGF processing is restricted by cell type, we infected a variety of mammalian cell lines with both recombinant viruses; all accumulated the same 35-kDa precursor and secreted NGF. Thus, many types of cells have the machinery to process and secrete NGF. However, NGF accumulated intracellularly (presumably in secretory granules) in cells with a regulated pathway of secretion (e.g., AtT-20 and HIT cells). In these cells, a membrane-permeable cyclic AMP analog, 8-bromo-cyclic AMP, stimulated NGF secretion. This suggests a mechanism for the regulation of NGF levels in which specific secretagogues, e.g., neurotransmitters, control NGF secretion.

scholarly journals Expression of the Catalytic Subunit of Human DNA Polymerase δ in Mammalian Cells Using a Vaccinia Virus Vector System

1995 ◽  
Vol 270 (14) ◽  
pp. 7993-7998 ◽  
Author(s):  
Peng Zhang ◽  
Isabelle Frugulhetti ◽  
Yunquan Jiang ◽  
Geraldine L. Holt ◽  
Richard C. Condit ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Dna Polymerase ◽  
Mammalian Cells ◽  
Virus Vector ◽  
Catalytic Subunit ◽  
Vector System ◽  
Dna Polymerase Δ ◽  
Human Dna ◽  
Vaccinia Virus Vector
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