Proteome‐Wide Anti–Hepatitis C Virus (HCV) and Anti‐HIV Antibody Profiling for Predicting and Monitoring the Response to HCV Therapy in HIV‐Coinfected Patients

Burbelo, Peter D.; Kovacs, Joseph A.; Ching, Kathryn H.; Issa, Alexandra T.; Iadarola, Michael J.; Murphy, Alison; Schlaak, Joerg F.; Masur, Henry; Polis, Michael A.; Kottilil, Shyam

doi:10.1086/655780

Cited by 19 publications

(25 citation statements)

References 15 publications

(18 reference statements)

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“…We chose to use serology for its advantages of tolerance for sequence divergence, a capacity to detect resolved as well as acute or active infections, and high sample throughput. We expected that like other RNA viruses, including HCV, different CHV variants would be genetically diverse, and we therefore synthesized antigen from the helicase domain of NS3, the most conserved viral protein in the genome and one that shows documented antigenicity in HCV (11).…”

Section: Discussionmentioning

confidence: 99%

“…1B). To examine possible antigenic cross-reactivity, we tested the equine seropositive and seronegative samples against helicase protein of HCV (11) and found all these samples to be nonreactive (Fig. 1C).…”

Section: Use Of the Lips To Search Chv-related Viruses In Other Animalmentioning

confidence: 99%

“…Similarly, although the demonstration of specific adaptive immune responses against viral structural and nonstructural proteins cannot in itself prove a causal relationship to disease, it does provide definitive evidence of host infection (2,7). Here, we describe the usefulness of a sensitive serological assay (7)(8)(9)(10)(11) that can be quickly established for new pathogens to screen different animal species for evidence of virus infection and thereby for identification of a novel virus's natural reservoir and host range. Our serology data indicated the presence of CHV-like viruses in horses and led to the genetic characterization of eight novel and genetically diverse nonprimate hepaciviruses (NPHVs).…”

mentioning

confidence: 99%

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Serology-Enabled Discovery of Genetically Diverse Hepaciviruses in a New Host

Burbelo

Dubovi

Simmonds

et al. 2012

J Virol

217

364

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Genetic and biological characterization of new hepaciviruses infecting animals contributes to our understanding of the ultimate origins of hepatitis C virus (HCV) infection in humans and dramatically enhances our ability to study its pathogenesis using tractable animal models. Animal homologs of HCV include a recently discovered canine hepacivirus (CHV) and GB virus B (GBV-B), both viruses with largely undetermined natural host ranges. Here we used a versatile serology-based approach to determine the natural host of the only known nonprimate hepacivirus (NPHV), CHV, which is also the closest phylogenetic relative of HCV. Recombinant protein expressed from the helicase domain of CHV NS3 was used as antigen in the luciferase immunoprecipitation system (LIPS) assay to screen several nonprimate animal species. Thirty-six samples from 103 horses were immunoreactive, and viral genomic RNA was present in 8 of the 36 seropositive animals and none of the seronegative animals. Complete genome sequences of these 8 genetically diverse NPHVs showed 14% (range, 6.4% to 17.2%) nucleotide sequence divergence, with most changes occurring at synonymous sites. RNA secondary structure prediction of the 383-base 5′ untranslated region of NPHV was refined and extended through mapping of polymorphic sites to unpaired regions or (semi)covariant pairings. Similar approaches were adopted to delineate extensive RNA secondary structures in the coding region of the genome, predicted to form 27 regularly spaced, thermodynamically stable stem-loops. Together, these findings suggest a promising new nonprimate animal model and provide a database that will aid creation of functional NPHV cDNA clones and other novel tools for hepacivirus studies.

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Section: Discussionmentioning

confidence: 99%

Section: Use Of the Lips To Search Chv-related Viruses In Other Animalmentioning

confidence: 99%

mentioning

confidence: 99%

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Serology-Enabled Discovery of Genetically Diverse Hepaciviruses in a New Host

Burbelo

Dubovi

Simmonds

et al. 2012

J Virol

217

364

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“…The wide dynamic range of antibody detection for many of these LIPS tests is advantageous for monitoring response to treatment. LIPS profiling of antibodies generated against multiple hepatitis C virus (HCV) proteins has lead to the identification of biomarkers for differentiating long-term responders for HCV treatment in HCV-HIV-coinfected individuals [12]. In these studies the combined antibody titers to three HCV proteins, core, ENV1 and NS4, had potential for identification of long term responders from relapsers at 40 weeks following treatment with interferon-α and ribavirin for HCV infection.…”

Section: Infectious Disease Diagnostics and Antigen Discovery By Lipsmentioning

confidence: 99%

Searching for biomarkers: humoral response profiling with luciferase immunoprecipitation systems

Burbelo¹,

Ching²,

Bren³

et al. 2011

Expert Review of Proteomics

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B-cell mediated humoral responses are triggered in many human diseases including autoimmune, cancer, neurologic, and infectious diseases. However, the full exploitation of the information contained within a patient's antibody repertoire, for diagnosis, monitoring and even disease prediction has been limited due to the poor diagnostic performance of many immunoassay formats. We have developed Luciferase immunoprecipitation systems (LIPS) that harnesses light emitting proteins to generate high definition antibody profiles optimal for both diagnostics and biomarker discovery. Here we describe the results and implications from a range of LIPS antibody profiling studies performed in our laboratory. These include highly sensitive diagnostics for domestic and global pathogens, insights into infection-related diseases, discovery of new biomarkers for human diseases, subcategorization of symptoms and identification of pathogenic autoantibodies against self-proteins. These investigations highlight the types of humoral response profiles associated with different diseases, provide new information related to disease pathogenesis, and provide a framework for incorporating LIPS antibody profiling into global health initiatives and disease monitoring.

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“…To date, several LIPS formats are available including single tube assays to measure one antigen in a single sample (Burbelo et al 2005), iterative testing using microtiter plates (Burbelo et al 2010b; Burbelo et al 2010c; Burbelo et al 2009d) and microwell arrays for simultaneously profiling antibody responses to panels of antigens (Burbelo et al 2011). In addition, a rapid format called QLIPS uses a relatively short incubation of 10 minutes but still shows sensitivity and specificity comparable to the standard 2 hour LIPS format (Burbelo et al 2009a; Burbelo et al 2009c; Burbelo et al 2008).…”

Section: Introductionmentioning

confidence: 99%

Microfluidic LIPS for serum antibody detection: demonstration of a rapid test for HSV-2 infection

Zubair

Burbelo

Vincent

et al. 2011

Biomed Microdevices

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There is great interest in point-of-care antibody testing for the diagnosis of infectious and autoimmune diseases. As a first step in the development of self-contained and miniaturized devices for highly quantitative antibody detection, we demonstrate the application of Luciferase Immunoprecipitation Systems (LIPS) technology in a microfluidic format. Protein A/G was immobilized on the walls of PDMS-glass microchannels of 500 nL volume. The assay proceeds with the simultaneous introduction of plasma and Renilla luciferase–tagged antigens. Following washing, coelenterazine substrate was added and bound antigen-luciferase measured by chemiluminescence. Total assay time, including rinsing and detection, is under ten minutes. Using these stable microfluidic devices, high diagnostic performance (100% sensitivity and 100% specificity) was achieved for the diagnosis of HSV-2 infection. Based on these findings, the LIPS microfluidic format should readily lend itself to automation and the transfer to portable instrumentation.

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Proteome‐Wide Anti–Hepatitis C Virus (HCV) and Anti‐HIV Antibody Profiling for Predicting and Monitoring the Response to HCV Therapy in HIV‐Coinfected Patients

Cited by 19 publications

References 15 publications

Serology-Enabled Discovery of Genetically Diverse Hepaciviruses in a New Host

Serology-Enabled Discovery of Genetically Diverse Hepaciviruses in a New Host

Searching for biomarkers: humoral response profiling with luciferase immunoprecipitation systems

Microfluidic LIPS for serum antibody detection: demonstration of a rapid test for HSV-2 infection

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