Dissection of the humoral immune response toward an immunodominant epitope of HIV: a model for the analysis of antibody diversity in HIV+ individuals

Enshell-Seijffers, David; Smelyanski, Larisa; Vardinon, Nurith; Yust, Israel; Gershoni, Jonathan M.

doi:10.1096/fj.00-0898com

Cited by 46 publications

(32 citation statements)

References 53 publications

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“…Although the vectors both have a pUC origin, the presence of different antibiotic selection markers ensured both were maintained in the short term for double display under chloramphenicol and ampicillin selection. This approach avoided virus genome engineering and is presumably limited only by the number of different resistance markers available for multiplex display on the same virus construct (18,20,21).…”

Section: Methodsmentioning

confidence: 99%

Viral assembly of oriented quantum dot nanowires

Mao

Flynn

Hayhurst

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

489

387

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The highly organized structure of M13 bacteriophage was used as an evolved biological template for the nucleation and orientation of semiconductor nanowires. To create this organized template, peptides were selected by using a pIII phage display library for their ability to nucleate ZnS or CdS nanocrystals. The successful peptides were expressed as pVIII fusion proteins into the crystalline capsid of the virus. The engineered viruses were exposed to semiconductor precursor solutions, and the resultant nanocrystals that were templated along the viruses to form nanowires were extensively characterized by using high-resolution analytical electron microscopy and photoluminescence. ZnS nanocrystals were well crystallized on the viral capsid in a hexagonal wurtzite or a cubic zinc blende structure, depending on the peptide expressed on the viral capsid. Electron diffraction patterns showed single-crystal type behavior from a polynanocrystalline area of the nanowire formed, suggesting that the nanocrystals on the virus were preferentially oriented with their [001] perpendicular to the viral surface. Peptides that specifically directed CdS nanocrystal growth were also engineered into the viral capsid to create wurtzite CdS virus-based nanowires. Lastly, heterostructured nucleation was achieved with a dual-peptide virus engineered to express two distinct peptides within the same viral capsid. This work represents a genetically controlled biological synthesis route to a semiconductor nanoscale heterostructure. E xploiting the biologically evolved, self-assembling structures of viral capsids is a possible mechanism in the ordering of technologically important materials. A key characteristic of materials synthesized by natural biological systems is the hierarchical organization of structures on many length scales with controlled size, shape, alignment, and orientation (1-8). There has been much interest in using biological templates in vitro to modulate the growth of inorganic, semiconductor, magnetic, and other technologically important materials (9-14). Our previous work, along with that of others, has shown that biological combinatorial approaches can identify amino acid sequences capable of interacting with a variety of materials (11,15). Here, we present a system that exploits the self-assembling, crystalline structure of the viral coat to nucleate and template II-VI semiconductor nanocrystals into highly oriented quantum dot nanowires. To accomplish this, the pVIII major coat protein was engineered to display evolutionary-selected peptides, enabling directed nanocrystal growth into viral-semiconductor hybrid nanowires that were crystallographically oriented over the micrometer size range.The helical major coat protein, pVIII, of viruses can be engineered to express fusion proteins that readily self-assemble into a highly oriented viral coat structure. A 5-fold axis of rotation relates symmetric points on the coat (16, 17). Peptide sequences engineered into the pVIII protein for use as nanocrystal templates were select...

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Section: Methodsmentioning

confidence: 99%

Viral assembly of oriented quantum dot nanowires

Mao

Flynn

Hayhurst

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

489

387

View full text Add to dashboard Cite

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“…Random peptide phage display has been used to dissect Ab responses in the context of primate immunodeficiency virus infections, and several studies have identified HIV-1 or simian-human immunodeficiency virus (SHIV)-specific peptides (10)(11)(12)(13)(14). We developed a novel epitope selection strategy, termed "protection-linked (PL) biopanning," which was designed to differentiate between protective vaccine-induced Ab responses and those that failed to protect.…”

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confidence: 99%

Novel Biopanning Strategy To Identify Epitopes Associated with Vaccine Protection

Bachler

Humbert

Palikuqi

et al. 2013

J Virol

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cIdentifying immune correlates of protection is important to develop vaccines against infectious diseases. We designed a novel, universally applicable strategy to profile the antibody (Ab) repertoire of protected vaccine recipients, using recombinant phages encoding random peptide libraries. The new approach, termed "protection-linked (PL) biopanning," probes the Ab paratopes of protected vaccinees versus those with vaccine failure. As proof of concept, we screened plasma samples from vaccinated rhesus macaques (RMs) that had completely resisted multiple mucosal challenges with R5-tropic simian-human immunodeficiency viruses (SHIVs). The animals had been immunized with a multicomponent vaccine (multimeric HIV-1 gp160, HIV-1 Tat, and SIV Gag-Pol particles). After PL biopanning, we analyzed the phagotopes selected for amino acid homologies; in addition to the expected Env mimotopes, one recurring motif reflected the neutralizing Ab epitope at the N terminus (NT) of HIV-1 Tat. Subsequent binding and functional assays indicated that anti-Tat NT Abs were present only in completely or partially protected RMs; peak viremia of the latter was inversely correlated with anti-Tat NT Ab titers. In contrast, highly viremic, unvaccinated controls did not develop detectable Abs against the same epitope. Based upon the protective effect observed in vivo, we suggest that Tat should be included in multicomponent HIV-1 vaccines. Our data highlight the power of the new PL-biopanning strategy to identify Ab responses with significant association to vaccine protection, regardless of the mechanism(s) or targets of the protective Abs. PL biopanning is also unbiased with regard to pathogens or disease model, making it a universal tool.T he design of a safe, effective vaccine is desirable to control the AIDS pandemic. In order to improve future vaccine candidates, it is important to understand the protective mechanism(s) leading to reduced virus acquisition (1). Thus, correlates of infection risk for the recent RV144 vaccine efficacy trial (2) are the subject of intense study (3,4). However, additional information can be gained by dissecting immune responses in biologically relevant animal models where immunization induced complete protection from immunodeficiency virus challenge(s) (5, 6).Recent results of active (7,8) and passive (9) immunization trials in humans and in nonhuman primates suggested that antihuman immunodeficiency virus type 1 (HIV-1) Env antibodies (Abs) can have protective roles even in the absence of virus neutralization, implying that other Ab-mediated mechanisms may be involved in preventing mucosal virus acquisition. Thus, the ability to profile Ab epitopes associated with protection might help to identify potential targets for future HIV-1 vaccines.Random peptide phage display has been used to dissect Ab responses in the context of primate immunodeficiency virus infections, and several studies have identified HIV-1 or simian-human immunodeficiency virus (SHIV)-specific peptides (10-14). We developed a novel epitope...

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“…Furthermore, the hydrophobic CDRH3 regions recognize lipids [7,15] and at least 2F5 and 4E10 bnAbs are also cross reactive with human proteins, thus suggesting that tolerance may limit anti-MPER responses [25,26]. All these limitations seem to favor the diversion of humoral immune responses towards other gp41 regions, in particular the external loop, which has been described as an immunodominant nonneutralizing region [27].…”

Section: Discussionmentioning

confidence: 99%

Anti-MPER Antibodies with Heterogeneous Neutralization Capacity Are Detectable in Most Untreated HIV Infected Individuals

Molinos-AlbertLuis

CarrilloJorge

CurriuMarta

et al. 2014

AIDS Research and Human Retroviruses

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Background: The MPER region of the HIV-1 envelope glycoprotein gp41 is targeted by broadly neutralizing antibodies. However, the localization of this epitope in a hydrophobic environment seems to hamper the elicitation of these antibodies in HIV infected individuals. We have quantified and characterized anti-MPER antibodies by ELISA and by flow cytometry using a collection of mini gp41-derived proteins expressed on the surface of 293T cells. Longitudinal plasma samples from 35 HIV-1 infected individuals were assayed for MPER recognition and MPER-dependent neutralizing capacity using HIV-2 viruses engrafted with HIV-1 MPER sequences.

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Dissection of the humoral immune response toward an immunodominant epitope of HIV: a model for the analysis of antibody diversity in HIV+ individuals

Cited by 46 publications

References 53 publications

Viral assembly of oriented quantum dot nanowires

Viral assembly of oriented quantum dot nanowires

Novel Biopanning Strategy To Identify Epitopes Associated with Vaccine Protection

Anti-MPER Antibodies with Heterogeneous Neutralization Capacity Are Detectable in Most Untreated HIV Infected Individuals

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