tetano
Editor, Senior Moderator
J Virol. 2016 Apr 20. pii: JVI.00452-16. [Epub ahead of print]
[h=1]Influenza Hemagglutinin Glycoproteins with Different N-glycan Patterns Activate Dendritic Cells in Vitro.[/h] Liu WC[SUP]1[/SUP], Lin YL[SUP]2[/SUP], Spearman M[SUP]3[/SUP], Cheng PY[SUP]2[/SUP], Butler M[SUP]3[/SUP], Wu SC[SUP]4[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Influenza HA N-glycans serve important regulatory roles in the control of virus virulence, antigenicity, receptor-binding specificity, and viral immune escape. Considered essential for controlling innate and adaptive immune responses against influenza virus infections, DCs trigger pro-inflammatory and adaptive immune responses in hosts. In this study, we engineered CHO cell lines expressing recombinant HA from pandemic H1, H5, and H7 influenza viruses. rH1HA, rH5HA and rH7HA proteins were obtained as wild type or in the presence of kifunensine (KIF), or further with Endo H treated (KIF+E) to generate single GlcNAc N-glycans, consisting of (i) terminally sialylated complex type N-glycans, (ii) high mannose-type N-glycans, and (iii) single-N-acetylglucosamine (GlcNAc)-type N-glycans. Our results show that high mannose-type and single GlcNAc-type N-glycans, but not complex-type N-glycans, are capable of inducing more active hIL12 p40, hIL12 p70 and hIL-10 production in human DCs. The significantly increased expression levels of the HLA-DR, CD40, CD83 and CD86, as well reduced endocytotic capacity in human DC were noted in the high mannose-type rH1HA and single GlcNAc-type rH1HA groups compared to the complex type N-glycans rH1HA group. Our data indicate that native avian rHA (H5N1 and H7N9) are more immunostimulatory than human rHA (pH1N1). The high-mannose type or single GlcNAc type N-glycans of both avian and human HA types are more stimulatory than the complex type N-glycans. The HA-stimulated DC activation was partially through mannose receptor(s). These results provide more understanding of the contribution of glycosylation of viral proteins to the immune responses and may have implications for vaccine development.
[h=4]IMPORTANCE:[/h] Influenza viruses trigger seasonal epidemics or pandemics with mild-to-severe consequences for human and poultry populations. DCs are the most potent professional antigen-presenting cells, which play a crucial role for the link of innate and adaptive immunity. In this study, we obtained stable-expression CHO cells to produce rH1HA, rH5HA and rH7HA proteins containing distinct N-glycans patterns. These rHA proteins, each with a distinct N-glycan pattern, were used to investigate interactions with mouse and human DCs. Our data indicate that native avian rHA (H5N1 and H7N9) are more immunostimulatory than human rHA (pH1N1). High mannose type- and single GlcNAc-type N-glycans were more effective than complex-type N-glycans in triggering mouse and human DC activation and maturation. We believe these results provide some useful information for influenza vaccine development regarding how influenza HA proteins with different types of N-glycans activate DCs.
Copyright ? 2016, American Society for Microbiology. All Rights Reserved.
PMID: 27099319 [PubMed - as supplied by publisher]
[h=1]Influenza Hemagglutinin Glycoproteins with Different N-glycan Patterns Activate Dendritic Cells in Vitro.[/h] Liu WC[SUP]1[/SUP], Lin YL[SUP]2[/SUP], Spearman M[SUP]3[/SUP], Cheng PY[SUP]2[/SUP], Butler M[SUP]3[/SUP], Wu SC[SUP]4[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Influenza HA N-glycans serve important regulatory roles in the control of virus virulence, antigenicity, receptor-binding specificity, and viral immune escape. Considered essential for controlling innate and adaptive immune responses against influenza virus infections, DCs trigger pro-inflammatory and adaptive immune responses in hosts. In this study, we engineered CHO cell lines expressing recombinant HA from pandemic H1, H5, and H7 influenza viruses. rH1HA, rH5HA and rH7HA proteins were obtained as wild type or in the presence of kifunensine (KIF), or further with Endo H treated (KIF+E) to generate single GlcNAc N-glycans, consisting of (i) terminally sialylated complex type N-glycans, (ii) high mannose-type N-glycans, and (iii) single-N-acetylglucosamine (GlcNAc)-type N-glycans. Our results show that high mannose-type and single GlcNAc-type N-glycans, but not complex-type N-glycans, are capable of inducing more active hIL12 p40, hIL12 p70 and hIL-10 production in human DCs. The significantly increased expression levels of the HLA-DR, CD40, CD83 and CD86, as well reduced endocytotic capacity in human DC were noted in the high mannose-type rH1HA and single GlcNAc-type rH1HA groups compared to the complex type N-glycans rH1HA group. Our data indicate that native avian rHA (H5N1 and H7N9) are more immunostimulatory than human rHA (pH1N1). The high-mannose type or single GlcNAc type N-glycans of both avian and human HA types are more stimulatory than the complex type N-glycans. The HA-stimulated DC activation was partially through mannose receptor(s). These results provide more understanding of the contribution of glycosylation of viral proteins to the immune responses and may have implications for vaccine development.
[h=4]IMPORTANCE:[/h] Influenza viruses trigger seasonal epidemics or pandemics with mild-to-severe consequences for human and poultry populations. DCs are the most potent professional antigen-presenting cells, which play a crucial role for the link of innate and adaptive immunity. In this study, we obtained stable-expression CHO cells to produce rH1HA, rH5HA and rH7HA proteins containing distinct N-glycans patterns. These rHA proteins, each with a distinct N-glycan pattern, were used to investigate interactions with mouse and human DCs. Our data indicate that native avian rHA (H5N1 and H7N9) are more immunostimulatory than human rHA (pH1N1). High mannose type- and single GlcNAc-type N-glycans were more effective than complex-type N-glycans in triggering mouse and human DC activation and maturation. We believe these results provide some useful information for influenza vaccine development regarding how influenza HA proteins with different types of N-glycans activate DCs.
Copyright ? 2016, American Society for Microbiology. All Rights Reserved.
PMID: 27099319 [PubMed - as supplied by publisher]