Abstract:
:We have demonstrated that a liposomal array of well-ordered trimers enhances B cell activation, germinal center formation, and the elicitation of tier-2 autologous neutralizing antibodies. Previously, we coupled well-ordered cleavage-independent NFL trimers via their C-terminal polyhistidine tails to nickel lipids integrated into the lipid bilayer. Despite favorable in vivo effects, concern remained over the potentially longer-term in vivo instability of noncovalent linkage of the trimers to the liposomes. Accordingly, we tested both cobalt coupling and covalent linkage of the trimers to the liposomes by reengineering the polyhistidine tail to include a free cysteine on each protomer of model BG505 NFL trimers to allow covalent linkage. Both cobalt and cysteine coupling resulted in a high-density array of NFL trimers that was stable in both 20% mouse serum and 100 mM EDTA, whereas the nickel-conjugated trimers were not stable under these conditions. Binding analysis and calcium flux with anti-Env-specific B cells confirmed that the trimers maintained conformational integrity following coupling. Following immunization of mice, serologic analysis demonstrated that the covalently coupled trimers elicited Env-directed antibodies in a manner statistically significantly improved compared to soluble trimers and nickel-conjugated trimers. Importantly, the covalent coupling not only enhanced gp120-directed responses compared to soluble trimers, it also completely eliminated antibodies directed to the C-terminal His tag located at the "bottom" of the spike. In contrast, soluble and noncovalent formats efficiently elicited anti-His tag antibodies. These data indicate that covalent linkage of well-ordered trimers to liposomes in high-density array displays multiple advantages in vitro and in vivoIMPORTANCE Enveloped viruses typically encode a surface-bound glycoprotein that mediates viral entry into host cells and is a primary target for vaccine design. Liposomes with modified lipid head groups have a unique feature of capturing and displaying antigens on their surfaces, mimicking the native pathogens. Our first-generation nickel-based liposomes captured HIV-1 Env glycoprotein trimers via a noncovalent linkage with improved efficacy over soluble glycoprotein in activating germinal center B cells and eliciting tier-2 autologous neutralizing antibodies. In this study, we report the development of second-generation cobalt- and maleimide-based liposomes that have improved in vitro stability over nickel-based liposomes. In particular, the maleimide liposomes captured HIV-1 Env trimers via a more stable covalent bond, resulting in enhanced germinal center B cell responses that generated higher antibody titers than the soluble trimers and liposome-bearing trimers via noncovalent linkages. We further demonstrate that covalent coupling prevents release of the trimers prior to recognition by B cells and masks a nonneutralizing determinant located at the bottom of the trimer.
journal_name
J Viroljournal_title
Journal of virologyauthors
Bale S,Goebrecht G,Stano A,Wilson R,Ota T,Tran K,Ingale J,Zwick MB,Wyatt RTdoi
10.1128/JVI.00443-17subject
Has Abstractpub_date
2017-07-27 00:00:00issue
16eissn
0022-538Xissn
1098-5514pii
JVI.00443-17journal_volume
91pub_type
杂志文章abstract::Rice yellow mottle virus is classified in five major serotypes; the molecular diversity of the coat protein (CP) is well established, but the amino acids involved in the recognition by discriminant monoclonal antibodies (MAbs) remain unknown. Reconstruction of a phylogenetic tree and sequence alignment of the CP gene ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.79.7.4407-4414.2005
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abstract::We have investigated the antigenicity of the C- and N-terminal halves of pIX of human adenovirus types 2 and 3 (Ad2 and Ad3) as well as their orientations in virions. We found that only the C-terminal halves of Ad2 pIX and Ad3 pIX reacted in a subgenus-specific manner by enzyme-linked immunosorbent assay and immunoblo...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.73.7.6182-6187.1999
更新日期:1999-07-01 00:00:00
abstract::The activity of human immunodeficiency virus Rev as a regulator of viral mRNA expression is tightly linked to its ability to shuttle between the nucleus and cytoplasm; these properties are conferred by a leucine-rich nuclear export signal (NES) and by an arginine-rich nuclear localization signal/RNA binding domain (NL...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.24.11899-11910.2000
更新日期:2000-12-01 00:00:00
abstract::The initiation of human immunodeficiency virus type 1 (HIV-1) reverse transcription occurs at a site in the viral RNA genome which is designated the primer-binding site (PBS). The HIV-1 PBS is an 18-nucleotide sequence that is complementary to the 3'-terminal 18 nucleotides of tRNA(3Lys), which is used as the primer f...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.10.6021-6029.1995
更新日期:1995-10-01 00:00:00
abstract::Eleven highly conserved proteins comprise the poxvirus entry-fusion complex (EFC). We focused on vaccinia virus (VACV) O3, a 35-amino acid, largely hydrophobic component of unknown specific function. Experimental evolution was carried out by blindly passaging a virus that was severely impaired in entry due to deletion...
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pub_type: 杂志文章
doi:10.1128/JVI.02228-20
更新日期:2021-01-27 00:00:00
abstract::Nonstructural protein 5A (NS5A) is a membrane-associated essential component of the hepatitis C virus (HCV) replication complex. An N-terminal amphipathic alpha helix mediates in-plane membrane association of HCV NS5A and at the same time is likely involved in specific protein-protein interactions required for the ass...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01279-06
更新日期:2007-03-01 00:00:00
abstract::Following respiratory syncytial virus (RSV) challenge, mice immunized with RSV G or with formalin-inactivated RSV (FI-RSV) exhibit severe disease associated with type 2 cytokine production and pulmonary eosinophilia. This has led to the proposal that the presence of RSV G is the factor in FI-RSV that induces disease-e...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.78.11.6024-6032.2004
更新日期:2004-06-01 00:00:00
abstract::We have previously shown that two inhibitors specific for cellular cyclin-dependent kinases (cdks), Roscovitine (Rosco) and Olomoucine (Olo), block the replication of herpes simplex virus (HSV). Based on these results, we demonstrated that HSV replication requires cellular cdks that are sensitive to these drugs (L. M....
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.5.2107-2120.2000
更新日期:2000-03-01 00:00:00
abstract::HeLa cell nuclei, isolated 17 h after infection with human adenovirus type 2 (Ad2), were treated with 200 mM ammonium sulfate. The extract (S200 fraction) contained 50 to 70% of the nonintegrated Ad2 DNA, which was in the form of nucleoprotein complexes. These complexes contained native, intact Ad2 DNA (with the excep...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.19.1.61-81.1976
更新日期:1976-07-01 00:00:00
abstract::cis-Acting elements in the viral genome RNA (vRNA) are essential for the translation, replication, and/or encapsidation of RNA viruses. In this study, a novel conserved cis-acting element was identified in the capsid-coding region of mosquito-borne flavivirus. The downstream of 5' cyclization sequence (5'CS) pseudokno...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00243-13
更新日期:2013-06-01 00:00:00
abstract::DNA sequences which have structural features suggestive of their functioning as an origin of lytic-phase DNA replication were previously identified in both human herpesvirus 6B strain Z29 [HHV-6B (Z29)] and in HHV-6A (U1102). Plasmid constructs containing the putative HHV-6B (Z29) oriLyt element were replicated after ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.67.12.7680-7683.1993
更新日期:1993-12-01 00:00:00
abstract::Historically, the adenoviral E3 region was found to be nonessential for viral replication in vitro. In addition, adenoviruses whose genome was more than approximately 105% the size of the native genome were inefficiently packaged. These profound observations were used experimentally to insert transgenes into the adeno...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.79.9.5455-5465.2005
更新日期:2005-05-01 00:00:00
abstract::HTG2 hamster cells produce a defective murine sarcoma virus lacking gp70 and, consequently, viral surface projections (knobs), but the lack of knobs appears to have no effect on intramembrane particle distribution. In addition, it has been noted that the core of the virus remains in the "immature" form as a result of ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.36.3.872-877.1980
更新日期:1980-12-01 00:00:00
abstract::Herpesviruses encode an essential, maturational serine protease whose catalytic domain, assemblin (28 kDa), is released by self-cleavage from a 74-kDa precursor (pPR, pUL80a). Although there is considerable information about the structure and enzymatic characteristics of assemblin, a potential pharmacologic target, co...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02821-06
更新日期:2007-04-01 00:00:00
abstract::Thanks to new technologies which enable rapid and unbiased screening for viral nucleic acids in clinical specimens, an impressive number of previously unknown viruses have recently been discovered. Two research groups independently identified a novel negative-strand RNA virus, now designated avian bornavirus (ABV), in...
journal_title:Journal of virology
pub_type: 杂志文章,评审
doi:10.1128/JVI.02567-09
更新日期:2010-07-01 00:00:00
abstract::Herpesvirus saimiri (HVS) is the prototype gamma-2 herpesvirus, which naturally infects the squirrel monkey Saimiri sciureus, causing an asymptomatic but persistent infection. The latent phase of gamma-2 herpesviruses is characterized by their ability to persist in a dividing cell population while expressing a limited...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02171-06
更新日期:2007-04-01 00:00:00
abstract::The avian adenovirus CELO is a promising vector for gene transfer applications. In order to study this potentiality, we developed an improved method for construction of adenovirus vectors in cosmids that was used to engineer the CELO genome. For all the recombinant viruses constructed by this method, the ability to pr...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.75.11.5288-5301.2001
更新日期:2001-06-01 00:00:00
abstract::The current therapy for hepatitis B and C is based on systemic administration of recombinant human alpha interferon (r-hIFN-alpha). However, systemic delivery of r-hIFN-alpha is associated with severe side effects, but more importantly, it is effective in only a small percentage of patients. In an effort to maximize I...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.10.4816-4823.2000
更新日期:2000-05-01 00:00:00
abstract::The human adenovirus type 2 (Ad2) mutant Ad2ts111 has previously been shown to contain two mutations which result in a complex phenotype. Ad2ts111 contains a single base change in the early region 1B (E1B) 19,000-molecular-weight (19K) coding region which yields a cyt deg phenotype and another defect which maps to the...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.57.3.883-892.1986
更新日期:1986-03-01 00:00:00
abstract:UNLABELLED:Chikungunya virus (CHIKV) is a member of a globally distributed group of arthritogenic alphaviruses that cause weeks to months of debilitating polyarthritis/arthralgia, which is often poorly managed with current treatments. Arthritic disease is usually characterized by high levels of the chemokine CCL2 and a...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.03364-13
更新日期:2014-06-01 00:00:00
abstract::Some of the properties of a new human papovavirus, BK, have been examined. Host range studies of BK virus (BKV) showed human cells to be more sensitive to infection than monkey cells; human fetal brain cells appear to be highly sensitive to BKV, with the production of extensive cytopathology characterized by cytoplasm...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.12.3.625-631.1973
更新日期:1973-09-01 00:00:00
abstract::The herpes simplex virus type 1 (HSV-1) alpha protein ICP27 regulates the transition between the delayed-early and late phases of the viral infection. Previous genetic analyses have suggested that the important functional domains of ICP27 map to its carboxyl-terminal half. One striking feature of the primary sequence ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.67.4.1778-1787.1993
更新日期:1993-04-01 00:00:00
abstract::Recombinant poxvirus vectors are undergoing intensive evaluation as vaccine candidates for a variety of infectious pathogens. Avipoxviruses, such as canarypox virus, are replication deficient in mammalian cells by virtue of a poorly understood species-specific restriction. Highly attenuated vaccinia virus strains such...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02654-06
更新日期:2007-07-01 00:00:00
abstract::A strategy involving polymerase chain reaction amplification of cDNAs was designed to study the expression of the hamster polyomavirus (HaPV) early region in HaPV-transformed rat fibroblasts, productively HaPV-infected cells, and HaPV-induced lymphoma. We identified three mRNAs resulting from alternative splicing of o...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.66.4.2495-2504.1992
更新日期:1992-04-01 00:00:00
abstract::Inhibition of host-directed gene expression by the matrix (M) protein of vesicular stomatitis virus (VSV) effectively blocks host antiviral responses, promotes virus replication, and disables the host cell. However, dendritic cells (DC) have the capacity to resist these effects and remain functional during VSV infecti...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00680-13
更新日期:2013-11-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.77.20.10799-10807.2003
更新日期:2003-10-01 00:00:00
abstract:UNLABELLED:Kaposi's sarcoma-associated herpesvirus (KSHV) typically displays two different phases in its life cycle, the default latent phase and the lytic phase. There is a short period of lytic gene expression in the early stage of KSHV primary infection. The factors involved in the shutdown process of lytic gene exp...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00596-14
更新日期:2014-07-01 00:00:00
abstract::Phosphorylation of the βC1 protein encoded by the betasatellite of tomato yellow leaf curl China virus (TYLCCNB-βC1) by SNF1-related protein kinase 1 (SnRK1) plays a critical role in defense of host plants against geminivirus infection in Nicotiana benthamiana However, how phosphorylation of TYLCCNB-βC1 impacts its pa...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00300-17
更新日期:2017-07-27 00:00:00
abstract::HeLa cells infected with human rhinovirus type 2 synthesize a mixture of single-and double-stranded ribonucleic acid (RNA). The RNA synthesized by the membrane-bound RNA polymerase complex in vitro is also a mixture of single- and double-stranded RNA, whereas the deoxycholate-treated RNA polymerase complex synthesized...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.10.1.93-98.1972
更新日期:1972-07-01 00:00:00
abstract::Translation of poliovirus RNA occurs by the binding of ribosomes to an internal segment of RNA sequence within the 5' untranslated region of the viral RNA. This region is predicted to consist of six domains (I to VI) that possess complex secondary and tertiary structures. Domain IV is a large region in which alteratio...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.7.4381-4389.1995
更新日期:1995-07-01 00:00:00