The H9N2 virus preferentially bound to SAα2,3Gal-resialylated CRBCs, whereas the human H1N1 and seasonal human H3N2 influenza virus preferentially bound to the SAα2,6Gal-resialylated CRBCs (Figure 3). Figure 3 Receptor specificity of virus strains. (A) Unmodified (left) and VCNA-treated CRBCs (right). (B) SAα2,6Gal-resialylated CRBCs hemagglutinate the H3N2 and pdmH1N1 viruses (left). SAα2,3Gal-resialylated CRBCs

hemagglutinate the H9N2 virus (right). Top: two hemagglutination units. Bottom: 1:2 dilution. siRNA-transduced respiratory cells were resistant to viral challenge A reduction in viral yield was seen in ST6GAL1 siRNA-transduced A549 cells challenged with the H3N2 and pdmH1N1 strains as compared with control cells (Figure 4A,B). Similar results were observed

for HBE and HEp-2 cells (Additional file 1: Figure S3). No differences Selumetinib were observed when cells were infected with the avian H9N2 virus (Figure 4C). Figure 4 ST6GAL1 siRNA-transduced respiratory cells resisted human influenza virus challenge and did not induce an interferon response. Transduced A549 cells were challenged with H3N2, pdmH1N1, or H9N2 viruses. (A) A reduction in viral yield was seen in ST6GAL1 siRNA-transduced cells infected with and pdmH1N1 (B) H3N2 influenza viruses. a P < 0.05. (C) Viral yield was not affected when cells were infected with the avian H9N2 virus. (D) Treatment with ST6GAL1 siRNAs resulted in a reduced capacity for viral replication during virus entry. a P < 0.05. (E) ELISAs were used to measure levels of IFN-β production following treatment with siRNAs. Inhibition of ST6GAL1 expression affects virus binding and www.selleckchem.com/products/CP-673451.html internalization Virus particles were abundant on the surface of A549 cells transfected with control siRNAs, and those infected with viruses (Figure 5A,B). However, there was a reduction in the number of bound

virus particles for cells treated with ST6GAL1 siRNAs (Figure 5C). The genome copy number of viruses was reduced following transfection of the various cell lines (A549, HBE, and HEp-2) with ST6GAL1 siRNAs (Figure 4D) Bumetanide prior to viral infection. Figure 5 Virus particle binding assays. Virus particles binding to the surface of untransfected cells (A, black arrow), and cells treated with control siRNAs (B, black arrow). The binding of virus particles to the cell surface was adversely affected by treating with ST6GAL1 siRNAs (C, black arrow). The tested siRNAs did not induce an interferon LY411575 in vitro response The expression of IFN-β in supernatants of siRNA-transfected cell lines (A549, HBE and HEp-2) was not detected. As a positive control, a long double-stranded RNA that is known to induce the expression of IFN-β was included (Figure 4E). Discussion In our study, we were able to demonstrate that down-regulation of the major influenza receptor, SAα2,6Gal, in respiratory epithelial cells was a promising approach to prevent viral entry and establishment of an infection.