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Blood group substances as potential therapeutic agents for the prevention and treatment of infection with noroviruses proving novel binding patterns in human tissues.

Yazawa S, Yokobori T, Ueta G, Ide M, Altan B, Thongprachum A, Nishimura T, Nakajima T, Kominato Y, Asao T, Saniabadi AR, Furukawa K, Kuwano H, Le Pendu J, Ushijima H - PLoS ONE (2014)

Bottom Line: Blood group-related glycans determining ABO and Lewis blood groups are known to function as attachment factors for most of the norovirus (NoV) strains.To identify binding specificity of each NoV, recombinant norovirus-like particles (VLPs) and human saliva samples with different ABO, Lewis phenotypes and secretor status have been commonly applied.Further, A, B and O(H) blood group substances prepared from porcine and squid tissues were found to be effective for preventing ABO blood group-specific binding of VLPs to both saliva and mucosa samples.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgical Science, Gunma University Graduate School of Medicine, Maebashi, Japan ; Tokushima Research Institute, Otsuka Pharmaceutical Co. Ltd., Tokushima, Japan.

ABSTRACT
Blood group-related glycans determining ABO and Lewis blood groups are known to function as attachment factors for most of the norovirus (NoV) strains. To identify binding specificity of each NoV, recombinant norovirus-like particles (VLPs) and human saliva samples with different ABO, Lewis phenotypes and secretor status have been commonly applied. When binding specificities of VLPs prepared from 16 different genotypes of NoVs in GI and GII genogroups were characterized in samples of human gastric mucosa compared to human saliva based on blood group phenotypes, considerable differences were observed for several strains. Novel binding specificities determined by an ELISA using preparations from human gastric mucosa were also ascertained by immunohistochemical analyses using human jejunal mucosa, widely believed to be susceptible to NoV infection. Further, A, B and O(H) blood group substances prepared from porcine and squid tissues were found to be effective for preventing ABO blood group-specific binding of VLPs to both saliva and mucosa samples. Therefore, these blood group substances might have potential for the prevention and treatment of NoV infection.

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Related in: MedlinePlus

Binding assay of 13 NoV-VLPs to human saliva samples.Binding assays with human saliva (n = 32) from individuals with different ABO and Lewis phenotypes and secretor status were conducted using three different samples each from ABO(H) and Lewis-positive (Lea+ and Leb+) individuals and a single sample each from Lewis-negative (secretor and non-secretor) ones. Experiments of each sample were performed twice in duplicate and mean±SD values per individual blood types are given on the abscissa in the graph indicating optical density at 450 nm. At least eight different binding patterns were determined. See the details in the Text. Leb, Le(a−b+)sec.; Lea, Le(a+b−)non-sec.; Le(-), Le(a−b−); Sec., secretor; Non-sec., non-secretor.
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pone-0089071-g002: Binding assay of 13 NoV-VLPs to human saliva samples.Binding assays with human saliva (n = 32) from individuals with different ABO and Lewis phenotypes and secretor status were conducted using three different samples each from ABO(H) and Lewis-positive (Lea+ and Leb+) individuals and a single sample each from Lewis-negative (secretor and non-secretor) ones. Experiments of each sample were performed twice in duplicate and mean±SD values per individual blood types are given on the abscissa in the graph indicating optical density at 450 nm. At least eight different binding patterns were determined. See the details in the Text. Leb, Le(a−b+)sec.; Lea, Le(a+b−)non-sec.; Le(-), Le(a−b−); Sec., secretor; Non-sec., non-secretor.

Mentions: Sixteen VLPs belonging to GI and GII genogroups were examined to determine their binding specificities using an ELISA plate coated with a panel of saliva samples from various ABO, Lewis blood group phenotypes and secretor status (Figure 2). Binding patterns of each VLP were deduced from the determinants (Figure 1) present in the saliva samples, and eight different binding patterns were determined resulting from assays with 12 VLPs (Table 1). Seven of the 16 VLPs showed very weak (GII.3, GII.12 and GII.15) or no clear (GI.8, GII.1, GII.5 and GII.14) binding pattern with all the saliva samples tested. Results from these VLPs except GI.8 VLP (GII.1, GII.5 and GII.14) were, therefore, not shown in Figure 2. Blood group activities of saliva samples used in this study were determined together with their ABO and Lewis blood group phenotypes and secretor status (Table 2). The VLP from GI.11 bound to all in a panel of saliva samples with different A, B, O and Lewis blood group phenotypes irrespective of their secretor statuses. No significant difference was observed between blood group phenotypes and secretor status. However, a significant decrease of binding was determined between this VLP and a saliva sample from an Oh Bombay individual (OD450 = 0.619±0.147, P<0.05) in which no A, B O(H) or Leb antigen but only a small amount of Lea antigen was secreted. More than 90% (92.857±3.487%, attenuation ratio) of the binding observed in GI.11 with saliva samples from A, B and OLe(a−b+) secretors was abolished after removal of glycans attached to their saliva samples by means of periodate oxidation (Figure 3). For further characterization of binding specificity, GI.11 VLP was examined using a series of saliva samples, following modification of their antigenic determinants by means of digestions with glycosidases. When saliva samples from A, B and OLe(a−b+) secretors were treated with α1,3N-acetylgalactosaminidase (C. teritium A), α1,3galactosidase (C. sporogenesis maebashi) and α1,2fucosidase (B. fulminans), respectively, their antigenic determinants were specifically destroyed resulting in decreases of GI.11 binding to each sample and attenuation ratios were tentatively calculated as 19.7, 12.3 and 86.6%, respectively. Likewise, treatments of saliva samples from OLe(a+b−) and OLe(a−b−) non-secretors with α1,3/4fucosidase and β1,3galactosidase, respectively, also caused diminished binding of GI.11 to them and their attenuation ratios were 28.4 and 8.5%, respectively. Therefore, binding of GI.11 to saliva involved type-specific glycans, and the data indicate that GI.11 has the broadest binding pattern of reactivity with not only di-fucosylated and mono-fucosylated A, B and O(H) but also Lea and Lec determinants as described below. VLPs from 3 NoVs were found to show rather broad binding patterns reacting with di-fucosylated and mono-fucosylated A, B and O(H) (GII.6, GII.13) and B, O(H) and Lea (GII.7) determinants, respectively. Of note, none of these strains could bind significantly to the saliva samples from non-secretor or Lewis negative individuals, indicating lack of recognition in absence of an α1, 2fucosyl residue (GII.6, GII.13) or both α1, 2 and α1, 4fucosyl residues (GII.7) on the Lec determinant (P<0.01). VLPs from the other NoVs seemed to show a gradually narrower reactivity to the same saliva samples. The VLPs from GII.4 and GII.15, and from GI.1 and GI.3 reacted significantly (P<0.01) with di-fucosylated and mono-fucosylated A, B and di-fucosylated and mono-fucosylated A, O(H) determinants over O(H) and B determinants, respectively. Further, four VLPs showed very specific binding patterns to di-fucosylated and mono-fucosylated A (GI.4), di-fucosylated and mono-fucosylated B (GII.2) (P<0.01), and di-fucosylated B (GII.3, GII.12) (P<0.05) determinants over the other ones, respectively.


Blood group substances as potential therapeutic agents for the prevention and treatment of infection with noroviruses proving novel binding patterns in human tissues.

Yazawa S, Yokobori T, Ueta G, Ide M, Altan B, Thongprachum A, Nishimura T, Nakajima T, Kominato Y, Asao T, Saniabadi AR, Furukawa K, Kuwano H, Le Pendu J, Ushijima H - PLoS ONE (2014)

Binding assay of 13 NoV-VLPs to human saliva samples.Binding assays with human saliva (n = 32) from individuals with different ABO and Lewis phenotypes and secretor status were conducted using three different samples each from ABO(H) and Lewis-positive (Lea+ and Leb+) individuals and a single sample each from Lewis-negative (secretor and non-secretor) ones. Experiments of each sample were performed twice in duplicate and mean±SD values per individual blood types are given on the abscissa in the graph indicating optical density at 450 nm. At least eight different binding patterns were determined. See the details in the Text. Leb, Le(a−b+)sec.; Lea, Le(a+b−)non-sec.; Le(-), Le(a−b−); Sec., secretor; Non-sec., non-secretor.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3928367&req=5

pone-0089071-g002: Binding assay of 13 NoV-VLPs to human saliva samples.Binding assays with human saliva (n = 32) from individuals with different ABO and Lewis phenotypes and secretor status were conducted using three different samples each from ABO(H) and Lewis-positive (Lea+ and Leb+) individuals and a single sample each from Lewis-negative (secretor and non-secretor) ones. Experiments of each sample were performed twice in duplicate and mean±SD values per individual blood types are given on the abscissa in the graph indicating optical density at 450 nm. At least eight different binding patterns were determined. See the details in the Text. Leb, Le(a−b+)sec.; Lea, Le(a+b−)non-sec.; Le(-), Le(a−b−); Sec., secretor; Non-sec., non-secretor.
Mentions: Sixteen VLPs belonging to GI and GII genogroups were examined to determine their binding specificities using an ELISA plate coated with a panel of saliva samples from various ABO, Lewis blood group phenotypes and secretor status (Figure 2). Binding patterns of each VLP were deduced from the determinants (Figure 1) present in the saliva samples, and eight different binding patterns were determined resulting from assays with 12 VLPs (Table 1). Seven of the 16 VLPs showed very weak (GII.3, GII.12 and GII.15) or no clear (GI.8, GII.1, GII.5 and GII.14) binding pattern with all the saliva samples tested. Results from these VLPs except GI.8 VLP (GII.1, GII.5 and GII.14) were, therefore, not shown in Figure 2. Blood group activities of saliva samples used in this study were determined together with their ABO and Lewis blood group phenotypes and secretor status (Table 2). The VLP from GI.11 bound to all in a panel of saliva samples with different A, B, O and Lewis blood group phenotypes irrespective of their secretor statuses. No significant difference was observed between blood group phenotypes and secretor status. However, a significant decrease of binding was determined between this VLP and a saliva sample from an Oh Bombay individual (OD450 = 0.619±0.147, P<0.05) in which no A, B O(H) or Leb antigen but only a small amount of Lea antigen was secreted. More than 90% (92.857±3.487%, attenuation ratio) of the binding observed in GI.11 with saliva samples from A, B and OLe(a−b+) secretors was abolished after removal of glycans attached to their saliva samples by means of periodate oxidation (Figure 3). For further characterization of binding specificity, GI.11 VLP was examined using a series of saliva samples, following modification of their antigenic determinants by means of digestions with glycosidases. When saliva samples from A, B and OLe(a−b+) secretors were treated with α1,3N-acetylgalactosaminidase (C. teritium A), α1,3galactosidase (C. sporogenesis maebashi) and α1,2fucosidase (B. fulminans), respectively, their antigenic determinants were specifically destroyed resulting in decreases of GI.11 binding to each sample and attenuation ratios were tentatively calculated as 19.7, 12.3 and 86.6%, respectively. Likewise, treatments of saliva samples from OLe(a+b−) and OLe(a−b−) non-secretors with α1,3/4fucosidase and β1,3galactosidase, respectively, also caused diminished binding of GI.11 to them and their attenuation ratios were 28.4 and 8.5%, respectively. Therefore, binding of GI.11 to saliva involved type-specific glycans, and the data indicate that GI.11 has the broadest binding pattern of reactivity with not only di-fucosylated and mono-fucosylated A, B and O(H) but also Lea and Lec determinants as described below. VLPs from 3 NoVs were found to show rather broad binding patterns reacting with di-fucosylated and mono-fucosylated A, B and O(H) (GII.6, GII.13) and B, O(H) and Lea (GII.7) determinants, respectively. Of note, none of these strains could bind significantly to the saliva samples from non-secretor or Lewis negative individuals, indicating lack of recognition in absence of an α1, 2fucosyl residue (GII.6, GII.13) or both α1, 2 and α1, 4fucosyl residues (GII.7) on the Lec determinant (P<0.01). VLPs from the other NoVs seemed to show a gradually narrower reactivity to the same saliva samples. The VLPs from GII.4 and GII.15, and from GI.1 and GI.3 reacted significantly (P<0.01) with di-fucosylated and mono-fucosylated A, B and di-fucosylated and mono-fucosylated A, O(H) determinants over O(H) and B determinants, respectively. Further, four VLPs showed very specific binding patterns to di-fucosylated and mono-fucosylated A (GI.4), di-fucosylated and mono-fucosylated B (GII.2) (P<0.01), and di-fucosylated B (GII.3, GII.12) (P<0.05) determinants over the other ones, respectively.

Bottom Line: Blood group-related glycans determining ABO and Lewis blood groups are known to function as attachment factors for most of the norovirus (NoV) strains.To identify binding specificity of each NoV, recombinant norovirus-like particles (VLPs) and human saliva samples with different ABO, Lewis phenotypes and secretor status have been commonly applied.Further, A, B and O(H) blood group substances prepared from porcine and squid tissues were found to be effective for preventing ABO blood group-specific binding of VLPs to both saliva and mucosa samples.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgical Science, Gunma University Graduate School of Medicine, Maebashi, Japan ; Tokushima Research Institute, Otsuka Pharmaceutical Co. Ltd., Tokushima, Japan.

ABSTRACT
Blood group-related glycans determining ABO and Lewis blood groups are known to function as attachment factors for most of the norovirus (NoV) strains. To identify binding specificity of each NoV, recombinant norovirus-like particles (VLPs) and human saliva samples with different ABO, Lewis phenotypes and secretor status have been commonly applied. When binding specificities of VLPs prepared from 16 different genotypes of NoVs in GI and GII genogroups were characterized in samples of human gastric mucosa compared to human saliva based on blood group phenotypes, considerable differences were observed for several strains. Novel binding specificities determined by an ELISA using preparations from human gastric mucosa were also ascertained by immunohistochemical analyses using human jejunal mucosa, widely believed to be susceptible to NoV infection. Further, A, B and O(H) blood group substances prepared from porcine and squid tissues were found to be effective for preventing ABO blood group-specific binding of VLPs to both saliva and mucosa samples. Therefore, these blood group substances might have potential for the prevention and treatment of NoV infection.

Show MeSH
Related in: MedlinePlus