Structural Basis for Norovirus Inhibition by Human Milk Oligosaccharides

Weichert, Stefan; Koromyslova, A.D.; Singh, B.K.; Hansman, Satoko; Jennewein, Stefan; Schroten, Horst; Hansman, Grant S.

doi:10.1128/jvi.03223-15

Cited by 125 publications

(135 citation statements)

References 27 publications

Supporting

Mentioning

129

Contrasting

Unclassified

Order By: Relevance

“…In addition, FUT2 functionality has been linked to infection with rotavirus, another enteric pathogen that causes diarrheal disease in children, and that utilizes cell surface carbohydrates for attachment (114). These results provide a basis for identifying individuals resistant to infection by these viruses and suggest that the development of purified fucose-containing oligosaccharides may be effective as anti-infectives (163). …”

Section: Norovirus and Rotavirusmentioning

confidence: 93%

Human Genetic Determinants of Viral Diseases

et al. 2017

View full text Add to dashboard Cite

Much progress has been made in the identification of specific human gene variants that contribute to enhanced susceptibility or resistance to viral diseases. Herein we review multiple discoveries made with genome-wide or candidate gene approaches that have revealed significant insights into virus–host interactions. Genetic factors that have been identified include genes encoding virus receptors, receptor-modifying enzymes, and a wide variety of innate and adaptive immunity-related proteins. We discuss a range of pathogenic viruses, including influenza virus, respiratory syncytial virus, human immunodeficiency virus, human T cell leukemia virus, human papilloma virus, hepatitis B and C viruses, herpes simplex virus, norovirus, rotavirus, parvovirus, and Epstein-Barr virus. Understanding the genetic underpinnings that affect infectious disease outcomes should allow tailored treatment and prevention approaches in the future.

show abstract

Section: Norovirus and Rotavirusmentioning

confidence: 93%

Human Genetic Determinants of Viral Diseases

et al. 2017

View full text Add to dashboard Cite

show abstract

“…Several studies showed that human milk or HMOs could compete with the HBGA binding sites on the norovirus capsid (27,28). We recently showed that two HMOs, 2=FL and 3FL, blocked norovirus from binding to HBGAs in a mostly dose-dependent manner (29). HMOs can resist degradation in the gut, and the majority is excreted intact in the feces (ϳ1% is absorbed by the infant).…”

Section: Human Norovirus and Hmosmentioning

confidence: 99%

“…The HMO binding site on the GII.10 capsid was recently discovered (29). The 2=FL and 3FL bound at the equivalent HBGA pocket on the norovirus capsid (Fig.…”

Section: Structural Basis For Norovirus Binding To Hmosmentioning

confidence: 99%

Human Norovirus Interactions with Histo-Blood Group Antigens and Human Milk Oligosaccharides

Schroten

Hanisch

Hansman

2016

J Virol

Self Cite

View full text Add to dashboard Cite

Human noroviruses interact with both human histo-blood group antigens (HBGAs) and human milk oligosaccharides (HMOs). The former are believed to be important for a virus infection, while the latter might act as natural decoys in the host during an infection. However, certain noroviruses are known to bind poorly to HBGAs and yet still cause infections; some interact with numerous HBGA types but are nonprevalent; and yet others bind HBGAs and seem to be increasing in prevalence. HBGAs and HMOs can be found as soluble antigens in humans, can be structurally alike, and can interact with equivalent residues at identical binding pockets on the capsid. In this Gem, we discuss HBGA and HMO binding studies for human noroviruses, concentrating on the clinically important genogroup II noroviruses. In short, the roles of HBGA and HMO interactions in norovirus infections are still unclear. Human noroviruses are the dominant cause of outbreaks of acute gastroenteritis. These viruses are also responsible for significant numbers of sporadic infections, are often found in asymptomatic cases, can cause chronic infections, and infect all age groups. Noroviruses are transmitted via the fecal-oral route, and infections often result from consumption of contaminated foods such as bivalves (clams and oysters). Noroviruses have a single-stranded, positive-sense RNA genome of ϳ7.5 kb. The genome contains three open reading frames (ORFs): ORF1 encodes nonstructural proteins, ORF2 encodes a capsid protein (VP1), and ORF3 encodes a minor capsid protein. Human norovirus is difficult to grow in cell cultures, and the exact locations of the cells and the cell type(s) that norovirus infect remain unknown. Expression of the capsid protein in insect cells can generate selfassembled virus-like particles (VLPs) that are morphologically and antigenically similar to the native virion (1). The norovirus particle is composed of 180 copies of VP1, which forms 90 VP1 homodimers. The X-ray crystal structure of norovirus VLPs identified two main domains, the shell (S) domain and the protruding (P) domain (2). The S domain surrounds the viral RNA, whereas the P domain is further subdivided into P1 and P2 subdomains and contains determinants for cell attachment and antigenicity. HUMAN NOROVIRUS AND HISTO-BLOOD GROUP ANTIGENSHuman noroviruses are known to interact with histo-blood group antigens (HBGAs), and this interaction could be important for infection (3-7). HBGAs can be found as soluble antigens in saliva and are expressed on the mucosal epithelium of the digestive tract. Therefore, noroviruses could interact several times with HBGAs during the course of an infection. Interestingly, human noroviruses can naturally bioaccumulate in bivalves, where they are thought to bind HBGA-like carbohydrates in the gastrointestinal epithelial cells (8). However, this accumulation does not appear to cause harm to the bivalves.Multiple gene families control human HBGA biosynthesis through the action of glycosyltransferases (9). As HBGAs can be either lipid or protein ...

show abstract

“…Pig gastric mucin (PGM) and saliva blocking assays were performed as previously described [69]. Briefly, ELISA plates were coated with 10 μg/ml PGM (Sigma, Germany) or with saliva type A or B diluted in PBS 1:2000.…”

Section: Blocking Assaysmentioning

confidence: 99%

Nanobodies Targeting Norovirus Capsid Reveal Functional Epitopes and Potential Mechanisms of Neutralization

Koromyslova

Hansman

2018

Biophysical Journal

Self Cite

View full text Add to dashboard Cite

Norovirus is the leading cause of gastroenteritis worldwide. Despite recent developments in norovirus propagation in cell culture, these viruses are still challenging to grow routinely. Moreover, little is known on how norovirus infects the host cells, except that histo-blood group antigens (HBGAs) are important binding factors for infection and cell entry. Antibodies that bind at the HBGA pocket and block attachment to HBGAs are believed to neutralize the virus. However, additional neutralization epitopes elsewhere on the capsid likely exist and impeding the intrinsic structural dynamics of the capsid could be equally important. In the current study, we investigated a panel of Nanobodies in order to probe functional epitopes that could trigger capsid rearrangement and/ or interfere with HBGA binding interactions. The precise binding sites of six Nanobodies (Nano-4, Nano-14, Nano-26, Nano-27, Nano-32, and Nano-42) were identified using X-ray crystallography. We showed that these Nanobodies bound on the top, side, and bottom of the norovirus protruding domain. The impact of Nanobody binding on norovirus capsid morphology was analyzed using electron microscopy and dynamic light scattering. We discovered that distinct Nanobody epitopes were associated with varied changes in particle structural integrity and assembly. Interestingly, certain Nanobody-induced capsid morphological changes lead to the capsid protein degradation and viral RNA exposure. Moreover, Nanobodies employed multiple inhibition mechanisms to prevent norovirus attachment to HBGAs, which included steric obstruction (Nano-14), allosteric interference (Nano-32), and violation of normal capsid morphology (Nano-26 and Nano-85). Finally, we showed that two Nanobodies (Nano-26 and Nano-85) not only compromised capsid integrity and inhibited VLPs attachment to HBGAs, but also recognized a broad panel of norovirus genotypes with high affinities. Consequently, Nano-26 and Nano-85 have a great potential to function as novel therapeutic agents against human noroviruses. PLOS Author summaryWe determined the binding sites of six novel human norovirus specific Nanobodies (Nano-4, Nano-14, Nano-26, Nano-27, Nano-32, and Nano-42) using X-ray crystallography. The unique Nanobody recognition epitopes were correlated with their potential neutralizing capacities. We showed that one Nanobody (Nano-26) bound numerous genogroup II genotypes and interacted with highly conserved capsid residues. Four Nanobodies (Nano-4, Nano-26, Nano-27, and Nano-42) bound to occluded regions on the intact particles and impaired normal capsid morphology and particle integrity. One Nanobody (Nano-14) bound contiguous to the HBGA pocket and interacted with several residues involved in binding HBGAs. We found that the Nanobodies delivered multiple inhibition mechanisms, which included steric obstruction, allosteric interference, and disruption of the capsid stability. Our data suggested that the HBGA pocket might not be an ideal target for drug development, since the surrounding regio...

show abstract

Structural Basis for Norovirus Inhibition by Human Milk Oligosaccharides

Cited by 125 publications

References 27 publications

Human Genetic Determinants of Viral Diseases

Human Genetic Determinants of Viral Diseases

Human Norovirus Interactions with Histo-Blood Group Antigens and Human Milk Oligosaccharides

Nanobodies Targeting Norovirus Capsid Reveal Functional Epitopes and Potential Mechanisms of Neutralization

Contact Info

Product

Resources

About