A consensus–hemagglutinin-based DNA vaccine that protects mice against divergent H5N1 influenza viruses

Chen, Ming-Wei; Cheng, Ting-Jen Rachel; Huang, Yaoxing; Jan, Jia-Tsrong; Ma, Shiou-Hwa; Yu, Alice L.; Wong, Chi‐Huey; Ho, David D.

doi:10.1073/pnas.0806901105

Cited by 138 publications

(128 citation statements)

References 37 publications

(46 reference statements)

Supporting

Mentioning

125

Contrasting

Order By: Relevance

“…However, it is not known how changes in glycosylation affect receptor-binding specificity and affinity, especially with regard to the most pathogenic H5N1 HA. To address this question, we have developed a glycan microarray comprising extensive structural analogs of the HA-binding ligand, and several defined glycoforms of HA were prepared by using the H5 consensus sequence (21) for quantitative binding analysis. Although previous studies have used HA from insect cell expression (16), glycosylation in insect cells differs from mammalian cells, with a marked difference being that complex type N-glycans terminating in galactose and sialic acid are not produced in insect cells.…”

Section: Creating Defined Ha Glycoforms For Quantitative Glycan Micromentioning

confidence: 99%

Glycans on influenza hemagglutinin affect receptor binding and immune response

Wang

Chen

Tseng

et al. 2009

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

Self Cite

283

256

View full text Add to dashboard Cite

Recent cases of avian influenza H5N1 and the swine-origin 2009 H1N1 have caused a great concern that a global disaster like the 1918 influenza pandemic may occur again. Viral transmission begins with a critical interaction between hemagglutinin (HA) glycoprotein, which is on the viral coat of influenza, and sialic acid (SA) containing glycans, which are on the host cell surface. To elucidate the role of HA glycosylation in this important interaction, various defined HA glycoforms were prepared, and their binding affinity and specificity were studied by using a synthetic SA microarray. Truncation of the N-glycan structures on HA increased SA binding affinities while decreasing specificity toward disparate SA ligands. The contribution of each monosaccharide and sulfate group within SA ligand structures to HA binding energy was quantitatively dissected. It was found that the sulfate group adds nearly 100-fold (2.04 kcal/mol) in binding energy to fully glycosylated HA, and so does the biantennary glycan to the monoglycosylated HA glycoform. Antibodies raised against HA protein bearing only a single N-linked GlcNAc at each glycosylation site showed better binding affinity and neutralization activity against influenza subtypes than the fully glycosylated HAs elicited. Thus, removal of structurally nonessential glycans on viral surface glycoproteins may be a very effective and general approach for vaccine design against influenza and other human viruses.flu vaccine ͉ glycan binding ͉ glycosylation T he highly pathogenic H5N1 and the 2009 swine-origin influenza A (H1N1) viruses have caused global outbreaks and raised a great concern that further changes in the viruses may occur to bring about a deadly pandemic (1, 2). Important contributions to our understanding of influenza infections have come from the studies on hemagglutinin (HA), a viral coat glycoprotein that binds to specific sialylated glycan receptors in the respiratory tract, allowing the virus to enter the cell (3-6). To cross the species barrier and infect the human population, avian HA must change its receptorbinding preference from a terminally sialylated glycan that contains ␣2,3 (avian)-linked to ␣2,6 (human)-linked sialic acid motifs (7), and this switch could occur through only two mutations, as in the 1918 pandemic (8). Understanding the factors that affect influenza binding to glycan receptors is thus critical for developing methods to control any future crossover influenza strains that have pandemic potential.HA is a homotrimeric transmembrane protein with an ectodomain composed of a globular head and a stem region (3). Both regions carry N-linked oligosaccharides (9), which affect the functional properties of HA (10, 11). Among different subtypes of influenza A viruses, there is extensive variation in the glycosylation sites of the head region, whereas the stem oligosaccharides are more conserved and required for fusion activity (11). Glycans near antigenic peptide epitopes interfere with antibody recognition (12), and glycans near the proteolytic ...

show abstract

Section: Creating Defined Ha Glycoforms For Quantitative Glycan Micromentioning

confidence: 99%

Glycans on influenza hemagglutinin affect receptor binding and immune response

Wang

Chen

Tseng

et al. 2009

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

Self Cite

283

256

View full text Add to dashboard Cite

show abstract

“…Another approach involves consensus sequences that combine many H5N1 hemagglu-tinin sequences into a single gene. Of these approaches, only the approach with consensus sequences has been shown to provide partial protection against a diverse panel of H5N1 isolates (22).…”

mentioning

confidence: 99%

Broad Protection against Avian Influenza Virus by Using a Modified Vaccinia Ankara Virus Expressing a Mosaic Hemagglutinin Gene

Kamlangdee

Kingstad-Bakke

Anderson

et al. 2014

J Virol

View full text Add to dashboard Cite

A critical failure in our preparedness for an influenza pandemic is the lack of a universal vaccine. Influenza virus strains diverge by 1 to 2% per year, and commercially available vaccines often do not elicit protection from one year to the next, necessitating frequent formulation changes. This represents a major challenge to the development of a cross-protective vaccine that can protect against circulating viral antigenic diversity. We have constructed a recombinant modified vaccinia virus Ankara (MVA) that expresses an H5N1 mosaic hemagglutinin (H5M) (MVA-H5M). This mosaic was generated in silico using 2,145 field-sourced H5N1 isolates. A single dose of MVA-H5M provided 100% protection in mice against clade 0, 1, and 2 avian influenza viruses and also protected against seasonal H1N1 virus (A/Puerto Rico/8/34). It also provided short-term (10 days) and long-term (6 months) protection postvaccination. Both neutralizing antibodies and antigen-specific CD4 ؉ and CD8 ؉ T cells were still detected at 5 months postvaccination, suggesting that MVA-H5M provides long-lasting immunity. IMPORTANCEInfluenza viruses infect a billion people and cause up to 500,000 deaths every year. A major problem in combating influenza is the lack of broadly effective vaccines. One solution from the field of human immunodeficiency virus vaccinology involves a novel in silico mosaic approach that has been shown to provide broad and robust protection against highly variable viruses. Unlike a consensus algorithm which picks the most frequent residue at each position, the mosaic method chooses the most frequent Tcell epitopes and combines them to form a synthetic antigen. These studies demonstrated that a mosaic influenza virus H5 hemagglutinin expressed by a viral vector can elicit full protection against diverse H5N1 challenges as well as induce broader immunity than a wild-type hemagglutinin.

show abstract

“…Consensus HA and neuraminidase (NA) synthetic DNAs have also been explored as an approach to enhance H5N1 vaccine efficacies. With such DNA vaccines, only partial protection was observed in mice challenged with HPAI H5N1 (9)(10)(11). Although promising, the consensus DNA methods underestimate the true diversity of the circulating viruses and are based on influenza vaccine platforms not yet approved for human use.…”

mentioning

confidence: 99%

Feasibility of reconstructed ancestral H5N1 influenza viruses for cross-clade protective vaccine development

Ducatez

Bahl

Griffin

et al. 2010

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

View full text Add to dashboard Cite

Since the reemergence of highly pathogenic H5N1 influenza viruses in humans in 2003, these viruses have spread throughout avian species in Asia, Europe, and Africa. Their sustained circulation has resulted in the evolution of phylogenetically diverse lineages. Viruses from these lineages show considerable antigenic variation, which has confounded vaccine planning efforts. We reconstructed ancestral protein sequences at several nodes of the hemagglutinin (HA) and neuraminidase (NA) gene phylogenies that represent ancestors to diverse H5N1 virus clades. By using the same methods that have been used to generate currently licensed inactivated H5N1 vaccines, we were able to produce a panel of replication competent influenza viruses containing synthesized HA and NA genes representing the reconstructed ancestral proteins. We identified two of these viruses that showed promising in vitro cross-reactivity with clade 1, 2.1, 2.2, 2.3.4, and 4 viruses. To confirm that vaccine antigens derived from these viruses were able to elicit functional antibodies following immunization, we created whole-virus vaccines and compared their protective efficacy versus that of antigens from positive control, naturally occurring, and broadly reactive H5N1 viruses. The ancestral viruses’ vaccines provided robust protection against morbidity and mortality in ferrets challenged with H5N1 strains from clades 1, 2.1, and 2.2 in a manner similar to those based on the control strains. These findings provide proof of principle that viable, computationally derived vaccine seed viruses can be constructed within the context of currently licensed vaccine platforms. Such technologies should be explored to enhance the cross reactivity and availability of H5N1 influenza vaccines.

show abstract

A consensus–hemagglutinin-based DNA vaccine that protects mice against divergent H5N1 influenza viruses

Cited by 138 publications

References 37 publications

Glycans on influenza hemagglutinin affect receptor binding and immune response

Glycans on influenza hemagglutinin affect receptor binding and immune response

Broad Protection against Avian Influenza Virus by Using a Modified Vaccinia Ankara Virus Expressing a Mosaic Hemagglutinin Gene

Feasibility of reconstructed ancestral H5N1 influenza viruses for cross-clade protective vaccine development

Contact Info

Product

Resources

About