scholarly journals Disulfide Bonds and Membrane Topology of the Vaccinia Virus A17L Envelope Protein

2000 ◽  
Vol 74 (5) ◽  
pp. 2438-2442 ◽  
Author(s):  
Tatiana Betakova ◽  
Bernard Moss
Keyword(s):  
Vaccinia Virus ◽  
Envelope Protein ◽  
Disulfide Bonds ◽  
Recombinant Vaccinia Virus ◽  
Open Reading Frame ◽  
Membrane Topology ◽  
Recombinant Vaccinia ◽  
Reading Frame ◽  
Virion Assembly ◽  
Intramolecular Disulfide Bond

ABSTRACT The envelope protein encoded by the vaccinia virus A17L open reading frame is essential for virion assembly. Our mutagenesis studies indicated that cysteines 101 and 121 form an intramolecular disulfide bond and that cysteine 178 forms an intermolecular disulfide linking two A17L molecules. This arrangement of disulfide bonds has important implications for the topology of the A17L protein and supports a two-transmembrane model in which cysteines 101 and 121 are intraluminal and cysteine 178 is cytoplasmic. The structure of the A17L protein, however, was not dependent on these disulfide bonds, as a recombinant vaccinia virus with all three cysteine codons mutated to serines retained infectivity.

scholarly journals Induction of a Mucosal Cytotoxic T-Lymphocyte Response by Intrarectal Immunization with a Replication-Deficient Recombinant Vaccinia Virus Expressing Human Immunodeficiency Virus 89.6 Envelope Protein

1998 ◽  
Vol 72 (10) ◽  
pp. 8264-8272 ◽  
Author(s):  
Igor M. Belyakov ◽  
Linda S. Wyatt ◽  
Jeffrey D. Ahlers ◽  
Patricia Earl ◽  
C. David Pendleton ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Vaccinia Virus ◽  
T Lymphocyte ◽  
Envelope Protein ◽  
Cytotoxic T Lymphocyte ◽  
Recombinant Vaccinia Virus ◽  
Mucosal Immunization ◽  
Recombinant Vaccinia ◽  
Immunodeficiency Virus ◽  
Hiv 1

ABSTRACT To improve the safety of recombinant vaccinia virus vaccines, modified vaccinia virus Ankara (MVA) has been employed, because it has a replication defect in most mammalian cells. Here we apply MVA to human immunodeficiency virus type 1 (HIV-1) vaccine development by incorporating the envelope protein gp160 of HIV-1 primary isolate strain 89.6 (MVA 89.6) and use it to induce mucosal cytotoxic-T-lymphocyte (CTL) immunity. In initial studies to define a dominant CTL epitope for HIV-1 89.6 gp160, we mapped the epitope to a sequence, IGPGRAFYAR (from the V3 loop), homologous to that recognized by HIV MN loop-specific CTL and showed that HIV-1 MN-specific CTLs cross-reactively recognize the corresponding epitope from strain 89.6 presented by H-2Dd. Having defined the CTL specificity, we immunized BALB/c mice intrarectally with recombinant MVA 89.6. A single mucosal immunization with MVA 89.6 was able to elicit long-lasting antigen-specific mucosal (Peyer’s patch and lamina propria) and systemic (spleen) CTL responses as effective as or more effective than those of a replication-competent vaccinia virus expressing 89.6 gp160. Immunization with MVA 89.6 led to (i) the loading of antigen-presenting cells in vivo, as measured by the ex vivo active presentation of the P18-89.6 peptide to an antigen-specific CTL line, and (ii) the significant production of the proinflammatory cytokines (interleukin-6 and tumor necrosis factor alpha) in the mucosal sites. These results indicate that nonreplicating recombinant MVA may be at least as effective for mucosal immunization as replicating recombinant vaccinia virus.

scholarly journals Dengue 2 Virus Envelope Protein Expressed by a Recombinant Vaccinia Virus Fails to Protect Monkeys against Dengue

1988 ◽  
Vol 69 (8) ◽  
pp. 1921-1929 ◽  
Author(s):  
V. Deubel ◽  
R. M. Kinney ◽  
J. J. Esposito ◽  
C. B. Cropp ◽  
A. V. Vorndam ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Envelope Protein ◽  
Recombinant Vaccinia Virus ◽  
Virus Envelope ◽  
Recombinant Vaccinia ◽  
Virus Envelope Protein

scholarly journals The Vaccinia Virus A9L Gene Encodes a Membrane Protein Required for an Early Step in Virion Morphogenesis

2000 ◽  
Vol 74 (20) ◽  
pp. 9701-9711 ◽  
Author(s):  
Wendy W. Yeh ◽  
Bernard Moss ◽  
Elizabeth J. Wolffe
Keyword(s):  
Vaccinia Virus ◽  
Early Stage ◽  
Protein A ◽  
Recombinant Vaccinia Virus ◽  
Proteolytic Processing ◽  
Transcriptional Analysis ◽  
Recombinant Vaccinia ◽  
Reading Frame ◽  
Virus Particles ◽  
Virion Morphogenesis

ABSTRACT The A9L open reading frame of vaccinia virus was predicted to encode a membrane-associated protein. A transcriptional analysis of the A9L gene indicated that it was expressed at late times in vaccinia virus-infected cells. Late expression, as well as virion membrane association, was demonstrated by the construction and use of a recombinant vaccinia virus encoding an A9L protein with a C-terminal epitope tag. Immunoelectron microscopy revealed that the A9L protein was associated with both immature and mature virus particles and was oriented in the membrane with its C terminus exposed on the virion surface. To determine whether the A9L protein functions in viral assembly or infectivity, we made a conditional-lethal inducible recombinant vaccinia virus. In the absence of inducer, A9L expression and virus replication were undetectable. Under nonpermissive conditions, viral late protein synthesis occurred, but maturational proteolytic processing was inhibited, and there was an accumulation of membrane-coated electron-dense bodies, crescents, and immature virus particles, many of which appeared abnormal. We concluded that the product of the A9L gene is a viral membrane-associated protein and functions at an early stage in virion morphogenesis.

Anti-tumoral Effect of Recombinant Vaccinia Virus through US Guided Injection in a Rabbit Model of Hepatic VX2 Carcinoma

2006 ◽  
Vol 54 (2) ◽  
pp. 103 ◽  
Author(s):  
Jong Young Oh ◽  
Byeong Ho Park ◽  
Myong Jin Kang ◽  
Jin Han Cho ◽  
Jong Cheol Choi ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Rabbit Model ◽  
Recombinant Vaccinia Virus ◽  
Recombinant Vaccinia ◽  
Vx2 Carcinoma

Characterization of a recombinant vaccinia virus expressing human melanoma-associated antigen p97.

1988 ◽  
Vol 62 (1) ◽  
pp. 176-180 ◽  
Author(s):  
S L Hu ◽  
G D Plowman ◽  
P Sridhar ◽  
U S Stevenson ◽  
J P Brown ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Recombinant Vaccinia Virus ◽  
Human Melanoma ◽  
Recombinant Vaccinia
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