Limits...
Enterovirus 71 virion-associated galectin-1 facilitates viral replication and stability.

Lee PH, Liu CM, Ho TS, Tsai YC, Lin CC, Wang YF, Chen YL, Yu CK, Wang SM, Liu CC, Shiau AL, Lei HY, Chang CP - PLoS ONE (2015)

Bottom Line: In this study, we found that the serum levels of galectin-1 in EV71-infected children were higher than those in non-infected people.In EV71 infected cells, galectin-1 was found to be associated with the EV71 VP1 and VP3 via carbohydrate residues and subsequently released and bound to another cell surface along with the virus.Taken together, our findings uncover a new role of galectin-1 in facilitating EV71 virus infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology & Immunology, College of Medicine, National Cheng Kung University, Tainan, 701, Taiwan.

ABSTRACT
Enterovirus 71 (EV71) infection causes a myriad of diseases from mild hand-foot-and-mouth disease or herpangina to fatal brain stem encephalitis complicated with pulmonary edema. Several severe EV71 endemics have occurred in Asia-Pacific region, including Taiwan, and have become a serious threat to children's health. EV71 infection is initiated by the attachment of the virion to the target cell surface. Although this process relies primarily upon interaction between viruses and cell surface receptors, soluble factors may also influence the binding of EV71 to host cells. Galectin-1 has been reported to participate in several virus infections, but is not addressed in EV71. In this study, we found that the serum levels of galectin-1 in EV71-infected children were higher than those in non-infected people. In EV71 infected cells, galectin-1 was found to be associated with the EV71 VP1 and VP3 via carbohydrate residues and subsequently released and bound to another cell surface along with the virus. EV71 propagated from galectin-1 knockdown SK-N-SH cells exhibited lower infectivity in cultured cells and less pathogenicity in mice than the virus propagated from parental cells. In addition, this galectin-1-free EV71 virus was sensitive to high temperature and lost its viability after long-term storage, which could be restored following supplement of recombinant galectin-1. Taken together, our findings uncover a new role of galectin-1 in facilitating EV71 virus infection.

Show MeSH

Related in: MedlinePlus

Galectin-1-/- EV71 shows less cellular binding, infectivity, mice neurological syndromes, and mortality.(A) Galectin-1-/- EV71 viruses reduce their cell binding activity. SK-N-SH cells were incubated with WT, galectin-1 -/- EV71 or galectin-1 -/- EV71 viruses with recombinant galectin-1 (25 ng/ml) at 4°C (MOI = 100) for 3 h. The surface-bound EV71 was detected by anti-EV71 antibody and hence analyzed by flow cytometry. *p<0.05 by one-way ANOVA followed by Tukey correction.(B) Galectin-1-/- EV71 viruses show lower infectivity than WT viruses. SK-N-SH cells were infected with WT, galectin-1 -/- EV71 or galectin-1 -/- EV71 viruses with recombinant galectin-1 (25 ng/ml) at 37°C (MOI = 0.5) for 12 h. The EV71-infected cells were analyzed by anti-EV71 antibody via flow cytometry. The virus-induced cytopathic effect was monitored under a microscope. *p<0.05 by one-way ANOVA followed by Tukey correction.(C) Galectin-1-/- EV71 viruses cause less neurological syndromes and mortality in mice. One week-old C56BL/6 mice were infected with 106 PFU of WT (n = 7) or galectin-1 -/- EV71 (n = 6) by intraperitoneal injection. The clinical scores and survival rate were monitored daily post-infection. ** p<0.01 by the log-rank (Mantel-Cox) test. Results are representative of two (A, B) or three (C) independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4338065&req=5

pone.0116278.g005: Galectin-1-/- EV71 shows less cellular binding, infectivity, mice neurological syndromes, and mortality.(A) Galectin-1-/- EV71 viruses reduce their cell binding activity. SK-N-SH cells were incubated with WT, galectin-1 -/- EV71 or galectin-1 -/- EV71 viruses with recombinant galectin-1 (25 ng/ml) at 4°C (MOI = 100) for 3 h. The surface-bound EV71 was detected by anti-EV71 antibody and hence analyzed by flow cytometry. *p<0.05 by one-way ANOVA followed by Tukey correction.(B) Galectin-1-/- EV71 viruses show lower infectivity than WT viruses. SK-N-SH cells were infected with WT, galectin-1 -/- EV71 or galectin-1 -/- EV71 viruses with recombinant galectin-1 (25 ng/ml) at 37°C (MOI = 0.5) for 12 h. The EV71-infected cells were analyzed by anti-EV71 antibody via flow cytometry. The virus-induced cytopathic effect was monitored under a microscope. *p<0.05 by one-way ANOVA followed by Tukey correction.(C) Galectin-1-/- EV71 viruses cause less neurological syndromes and mortality in mice. One week-old C56BL/6 mice were infected with 106 PFU of WT (n = 7) or galectin-1 -/- EV71 (n = 6) by intraperitoneal injection. The clinical scores and survival rate were monitored daily post-infection. ** p<0.01 by the log-rank (Mantel-Cox) test. Results are representative of two (A, B) or three (C) independent experiments.

Mentions: In order to determine the role of virus-associated glaectin-1 on EV71 infection, we tried to generate galectin-1-/- EV71 virus from galectin-1-deficinet cells. Using a lentiviral vector containing galectin-1 shRNA, the galectin-1 was successfully silenced in SK-N-SH cells (Fig. 4A). The EV71 viruses propagated from galectin-1 silenced SK-N-SH cells showed similar viral structure as wild type viruses by electronic microscopy observation (S2 Fig.), and were then collected to detect virus-associated galectin-1. Compared to those from wild type cells, EV71 viruses propagated from glaectin-1 silenced cells did not contain any galectin-1 protein, as confirmed by both antibody-based precipitation assay and ELISA (Fig. 4B and C). Next, we proceeded by examining the infectivity and virulence of the galectin-1-free EV71 viruses. We found that galecetin-1-/- EV 71 viruses showed a lower binding ability to host cells compared with the wild type virus (Fig. 5A). They also infected fewer SK-N-SH cells and caused a milder cytopathic effect (Fig. 5B), indicating that the galectin-/- EV71 viruses presented reduced infectivity. Such a marked decline in cell binding and infectivity of galectin-1-/- EV71 viruses was recovered by a supplement of recombinant galectin-1 (Fig. 5A and B).To evaluate the virulence of the virus, one week-old C56BL/6 mice were infected with either wild type or galectin-1-/- EV71 viruses to monitor virus-caused mice neuropathological symptoms and mortality. The mice infected with wild type EV71 viruses showed initial clinical signs of hunchback and wasting from day 3 after infection and further suffered from limb weakness and paralysis with the progressing number of days, and almost all died by around day 9. However, those mice which were infected with galectin-1-/- EV71 viruses presented mild neuropathological symptoms and a high survival rate (70%) (Fig. 5C). These results indicate that EV71 virus-coupled galectin-1 can promote viral infectivity and virulence.


Enterovirus 71 virion-associated galectin-1 facilitates viral replication and stability.

Lee PH, Liu CM, Ho TS, Tsai YC, Lin CC, Wang YF, Chen YL, Yu CK, Wang SM, Liu CC, Shiau AL, Lei HY, Chang CP - PLoS ONE (2015)

Galectin-1-/- EV71 shows less cellular binding, infectivity, mice neurological syndromes, and mortality.(A) Galectin-1-/- EV71 viruses reduce their cell binding activity. SK-N-SH cells were incubated with WT, galectin-1 -/- EV71 or galectin-1 -/- EV71 viruses with recombinant galectin-1 (25 ng/ml) at 4°C (MOI = 100) for 3 h. The surface-bound EV71 was detected by anti-EV71 antibody and hence analyzed by flow cytometry. *p<0.05 by one-way ANOVA followed by Tukey correction.(B) Galectin-1-/- EV71 viruses show lower infectivity than WT viruses. SK-N-SH cells were infected with WT, galectin-1 -/- EV71 or galectin-1 -/- EV71 viruses with recombinant galectin-1 (25 ng/ml) at 37°C (MOI = 0.5) for 12 h. The EV71-infected cells were analyzed by anti-EV71 antibody via flow cytometry. The virus-induced cytopathic effect was monitored under a microscope. *p<0.05 by one-way ANOVA followed by Tukey correction.(C) Galectin-1-/- EV71 viruses cause less neurological syndromes and mortality in mice. One week-old C56BL/6 mice were infected with 106 PFU of WT (n = 7) or galectin-1 -/- EV71 (n = 6) by intraperitoneal injection. The clinical scores and survival rate were monitored daily post-infection. ** p<0.01 by the log-rank (Mantel-Cox) test. Results are representative of two (A, B) or three (C) independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4338065&req=5

pone.0116278.g005: Galectin-1-/- EV71 shows less cellular binding, infectivity, mice neurological syndromes, and mortality.(A) Galectin-1-/- EV71 viruses reduce their cell binding activity. SK-N-SH cells were incubated with WT, galectin-1 -/- EV71 or galectin-1 -/- EV71 viruses with recombinant galectin-1 (25 ng/ml) at 4°C (MOI = 100) for 3 h. The surface-bound EV71 was detected by anti-EV71 antibody and hence analyzed by flow cytometry. *p<0.05 by one-way ANOVA followed by Tukey correction.(B) Galectin-1-/- EV71 viruses show lower infectivity than WT viruses. SK-N-SH cells were infected with WT, galectin-1 -/- EV71 or galectin-1 -/- EV71 viruses with recombinant galectin-1 (25 ng/ml) at 37°C (MOI = 0.5) for 12 h. The EV71-infected cells were analyzed by anti-EV71 antibody via flow cytometry. The virus-induced cytopathic effect was monitored under a microscope. *p<0.05 by one-way ANOVA followed by Tukey correction.(C) Galectin-1-/- EV71 viruses cause less neurological syndromes and mortality in mice. One week-old C56BL/6 mice were infected with 106 PFU of WT (n = 7) or galectin-1 -/- EV71 (n = 6) by intraperitoneal injection. The clinical scores and survival rate were monitored daily post-infection. ** p<0.01 by the log-rank (Mantel-Cox) test. Results are representative of two (A, B) or three (C) independent experiments.
Mentions: In order to determine the role of virus-associated glaectin-1 on EV71 infection, we tried to generate galectin-1-/- EV71 virus from galectin-1-deficinet cells. Using a lentiviral vector containing galectin-1 shRNA, the galectin-1 was successfully silenced in SK-N-SH cells (Fig. 4A). The EV71 viruses propagated from galectin-1 silenced SK-N-SH cells showed similar viral structure as wild type viruses by electronic microscopy observation (S2 Fig.), and were then collected to detect virus-associated galectin-1. Compared to those from wild type cells, EV71 viruses propagated from glaectin-1 silenced cells did not contain any galectin-1 protein, as confirmed by both antibody-based precipitation assay and ELISA (Fig. 4B and C). Next, we proceeded by examining the infectivity and virulence of the galectin-1-free EV71 viruses. We found that galecetin-1-/- EV 71 viruses showed a lower binding ability to host cells compared with the wild type virus (Fig. 5A). They also infected fewer SK-N-SH cells and caused a milder cytopathic effect (Fig. 5B), indicating that the galectin-/- EV71 viruses presented reduced infectivity. Such a marked decline in cell binding and infectivity of galectin-1-/- EV71 viruses was recovered by a supplement of recombinant galectin-1 (Fig. 5A and B).To evaluate the virulence of the virus, one week-old C56BL/6 mice were infected with either wild type or galectin-1-/- EV71 viruses to monitor virus-caused mice neuropathological symptoms and mortality. The mice infected with wild type EV71 viruses showed initial clinical signs of hunchback and wasting from day 3 after infection and further suffered from limb weakness and paralysis with the progressing number of days, and almost all died by around day 9. However, those mice which were infected with galectin-1-/- EV71 viruses presented mild neuropathological symptoms and a high survival rate (70%) (Fig. 5C). These results indicate that EV71 virus-coupled galectin-1 can promote viral infectivity and virulence.

Bottom Line: In this study, we found that the serum levels of galectin-1 in EV71-infected children were higher than those in non-infected people.In EV71 infected cells, galectin-1 was found to be associated with the EV71 VP1 and VP3 via carbohydrate residues and subsequently released and bound to another cell surface along with the virus.Taken together, our findings uncover a new role of galectin-1 in facilitating EV71 virus infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology & Immunology, College of Medicine, National Cheng Kung University, Tainan, 701, Taiwan.

ABSTRACT
Enterovirus 71 (EV71) infection causes a myriad of diseases from mild hand-foot-and-mouth disease or herpangina to fatal brain stem encephalitis complicated with pulmonary edema. Several severe EV71 endemics have occurred in Asia-Pacific region, including Taiwan, and have become a serious threat to children's health. EV71 infection is initiated by the attachment of the virion to the target cell surface. Although this process relies primarily upon interaction between viruses and cell surface receptors, soluble factors may also influence the binding of EV71 to host cells. Galectin-1 has been reported to participate in several virus infections, but is not addressed in EV71. In this study, we found that the serum levels of galectin-1 in EV71-infected children were higher than those in non-infected people. In EV71 infected cells, galectin-1 was found to be associated with the EV71 VP1 and VP3 via carbohydrate residues and subsequently released and bound to another cell surface along with the virus. EV71 propagated from galectin-1 knockdown SK-N-SH cells exhibited lower infectivity in cultured cells and less pathogenicity in mice than the virus propagated from parental cells. In addition, this galectin-1-free EV71 virus was sensitive to high temperature and lost its viability after long-term storage, which could be restored following supplement of recombinant galectin-1. Taken together, our findings uncover a new role of galectin-1 in facilitating EV71 virus infection.

Show MeSH
Related in: MedlinePlus