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Entry Inhibition of Influenza Viruses with High Mannose Binding Lectin ESA-2 from the Red Alga Eucheuma serra through the Recognition of Viral Hemagglutinin.

Sato Y, Morimoto K, Kubo T, Sakaguchi T, Nishizono A, Hirayama M, Hori K - Mar Drugs (2015)

Bottom Line: Besides ESA-2, HM-binding plant lectin ConA, fucose-binding lectins such as fungal AOL from Aspergillus oryzae and AAL from Aleuria aurantia were active against H1N1-2009, but the potency of inhibition was of less magnitude compared with ESA-2.Direct interaction between ESA-2 and a viral envelope glycoprotein, hemagglutinin (HA), was demonstrated by ELISA assay.Upon treatment with ESA-2, no viral antigens were detected in the host cells, indicating that ESA-2 inhibited the initial steps of virus entry into the cells.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Pharmacy, Yasuda Women's University, 6-13-1 Yasuhigashi, Asaminami-Ku, Hiroshima 731-0153, Japan. sato-y@yasuda-u.ac.jp.

ABSTRACT
Lectin sensitivity of the recent pandemic influenza A virus (H1N1-2009) was screened for 12 lectins with various carbohydrate specificity by a neutral red dye uptake assay with MDCK cells. Among them, a high mannose (HM)-binding anti-HIV lectin, ESA-2 from the red alga Eucheuma serra, showed the highest inhibition against infection with an EC50 of 12.4 nM. Moreover, ESA-2 exhibited a wide range of antiviral spectrum against various influenza strains with EC50s of pico molar to low nanomolar levels. Besides ESA-2, HM-binding plant lectin ConA, fucose-binding lectins such as fungal AOL from Aspergillus oryzae and AAL from Aleuria aurantia were active against H1N1-2009, but the potency of inhibition was of less magnitude compared with ESA-2. Direct interaction between ESA-2 and a viral envelope glycoprotein, hemagglutinin (HA), was demonstrated by ELISA assay. This interaction was effectively suppressed by glycoproteins bearing HM-glycans, indicating that ESA-2 binds to the HA of influenza virus through HM-glycans. Upon treatment with ESA-2, no viral antigens were detected in the host cells, indicating that ESA-2 inhibited the initial steps of virus entry into the cells. ESA-2 would thus be useful as a novel microbicide to prevent penetration of viruses such as HIV and influenza viruses to the host cells.

No MeSH data available.


Related in: MedlinePlus

Effect of ESA-2 on viral infection in the presence of mucins or human saliva. H292 cells were infected with A/Udorn/72 for 24 h in the presence or absence of 200 nM ESA-2 in the DMEM containing 10 µg/mL of mucin type I (bovine submaxillary gland), mucin type III (porcine stomach), or human saliva (10× dilution). Cell viability was evaluated by amide black staining followed by quantitation by image processing. The data was shown as percentage of cell viability (mock infection; 100%, virus infection; 0%).
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marinedrugs-13-03454-f005: Effect of ESA-2 on viral infection in the presence of mucins or human saliva. H292 cells were infected with A/Udorn/72 for 24 h in the presence or absence of 200 nM ESA-2 in the DMEM containing 10 µg/mL of mucin type I (bovine submaxillary gland), mucin type III (porcine stomach), or human saliva (10× dilution). Cell viability was evaluated by amide black staining followed by quantitation by image processing. The data was shown as percentage of cell viability (mock infection; 100%, virus infection; 0%).

Mentions: Direct interaction between ESA-2 and influenza envelope HA glycoproteins was demonstrated in ELISA assay (Figure 4). HA was dose-dependently bound to immobilized ESA-2 on the plate but not to the reference glycoprotein. This interaction was competitively inhibited by yeast mannan, a selective inhibitor of ESA-2, indicating ESA-2 bound to the HA through HM glycans. It has been demonstrated that CV-N exhibited anti-influenza activity by directly binding to HA1 [18]. Anti-influenza activity of ESA-2 exemplified in this study seems to be the same mechanism as that of CV-N. However, despite partially overlapping carbohydrate specificity for HM-glycans, CV-N and other HM-binding lectins, MVL from Microcystis viridis and GNA from Galanthus nivalis inhibited HCV infection through a different and complex mode of action [27]. In this respect, interaction of ESA-2 with cellular proteins, which potentially leads to various cellular responses such as proliferation or inflammation, should be taken into account in undertaking in vivo application. To evaluate the effectiveness of ESA-2 under conditions that model natural environment, we performed in vitro infection assays in the presence of mucins or human saliva. In this test, H292 cells were infected with A/Udorn/72 for 24 h and the cell viability was determined. As shown in Figure 5, 200 nM ESA-2 efficiently inhibited viral infection in the presence of mucin type I (bovine submaxillary gland), mucin type III (porcine stomach), and human saliva, although the saliva by itself inhibited the viral infection moderately.


Entry Inhibition of Influenza Viruses with High Mannose Binding Lectin ESA-2 from the Red Alga Eucheuma serra through the Recognition of Viral Hemagglutinin.

Sato Y, Morimoto K, Kubo T, Sakaguchi T, Nishizono A, Hirayama M, Hori K - Mar Drugs (2015)

Effect of ESA-2 on viral infection in the presence of mucins or human saliva. H292 cells were infected with A/Udorn/72 for 24 h in the presence or absence of 200 nM ESA-2 in the DMEM containing 10 µg/mL of mucin type I (bovine submaxillary gland), mucin type III (porcine stomach), or human saliva (10× dilution). Cell viability was evaluated by amide black staining followed by quantitation by image processing. The data was shown as percentage of cell viability (mock infection; 100%, virus infection; 0%).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4483639&req=5

marinedrugs-13-03454-f005: Effect of ESA-2 on viral infection in the presence of mucins or human saliva. H292 cells were infected with A/Udorn/72 for 24 h in the presence or absence of 200 nM ESA-2 in the DMEM containing 10 µg/mL of mucin type I (bovine submaxillary gland), mucin type III (porcine stomach), or human saliva (10× dilution). Cell viability was evaluated by amide black staining followed by quantitation by image processing. The data was shown as percentage of cell viability (mock infection; 100%, virus infection; 0%).
Mentions: Direct interaction between ESA-2 and influenza envelope HA glycoproteins was demonstrated in ELISA assay (Figure 4). HA was dose-dependently bound to immobilized ESA-2 on the plate but not to the reference glycoprotein. This interaction was competitively inhibited by yeast mannan, a selective inhibitor of ESA-2, indicating ESA-2 bound to the HA through HM glycans. It has been demonstrated that CV-N exhibited anti-influenza activity by directly binding to HA1 [18]. Anti-influenza activity of ESA-2 exemplified in this study seems to be the same mechanism as that of CV-N. However, despite partially overlapping carbohydrate specificity for HM-glycans, CV-N and other HM-binding lectins, MVL from Microcystis viridis and GNA from Galanthus nivalis inhibited HCV infection through a different and complex mode of action [27]. In this respect, interaction of ESA-2 with cellular proteins, which potentially leads to various cellular responses such as proliferation or inflammation, should be taken into account in undertaking in vivo application. To evaluate the effectiveness of ESA-2 under conditions that model natural environment, we performed in vitro infection assays in the presence of mucins or human saliva. In this test, H292 cells were infected with A/Udorn/72 for 24 h and the cell viability was determined. As shown in Figure 5, 200 nM ESA-2 efficiently inhibited viral infection in the presence of mucin type I (bovine submaxillary gland), mucin type III (porcine stomach), and human saliva, although the saliva by itself inhibited the viral infection moderately.

Bottom Line: Besides ESA-2, HM-binding plant lectin ConA, fucose-binding lectins such as fungal AOL from Aspergillus oryzae and AAL from Aleuria aurantia were active against H1N1-2009, but the potency of inhibition was of less magnitude compared with ESA-2.Direct interaction between ESA-2 and a viral envelope glycoprotein, hemagglutinin (HA), was demonstrated by ELISA assay.Upon treatment with ESA-2, no viral antigens were detected in the host cells, indicating that ESA-2 inhibited the initial steps of virus entry into the cells.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Pharmacy, Yasuda Women's University, 6-13-1 Yasuhigashi, Asaminami-Ku, Hiroshima 731-0153, Japan. sato-y@yasuda-u.ac.jp.

ABSTRACT
Lectin sensitivity of the recent pandemic influenza A virus (H1N1-2009) was screened for 12 lectins with various carbohydrate specificity by a neutral red dye uptake assay with MDCK cells. Among them, a high mannose (HM)-binding anti-HIV lectin, ESA-2 from the red alga Eucheuma serra, showed the highest inhibition against infection with an EC50 of 12.4 nM. Moreover, ESA-2 exhibited a wide range of antiviral spectrum against various influenza strains with EC50s of pico molar to low nanomolar levels. Besides ESA-2, HM-binding plant lectin ConA, fucose-binding lectins such as fungal AOL from Aspergillus oryzae and AAL from Aleuria aurantia were active against H1N1-2009, but the potency of inhibition was of less magnitude compared with ESA-2. Direct interaction between ESA-2 and a viral envelope glycoprotein, hemagglutinin (HA), was demonstrated by ELISA assay. This interaction was effectively suppressed by glycoproteins bearing HM-glycans, indicating that ESA-2 binds to the HA of influenza virus through HM-glycans. Upon treatment with ESA-2, no viral antigens were detected in the host cells, indicating that ESA-2 inhibited the initial steps of virus entry into the cells. ESA-2 would thus be useful as a novel microbicide to prevent penetration of viruses such as HIV and influenza viruses to the host cells.

No MeSH data available.


Related in: MedlinePlus