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Identification of novel viral receptors with cell line expressing viral receptor-binding protein.

Mei M, Ye J, Qin A, Wang L, Hu X, Qian K, Shao H - Sci Rep (2015)

Bottom Line: Thus, the viral cell receptor is an attractive target for anti-viral strategies, and the identification of viral cell receptor is critical for better understanding and controlling viral disease.Our data demonstrate that antibodies or siRNA to chANXA2 significantly inhibited ALV-J infection and replication, and over-expression of chANXA2 permitted the entry of ALV-J into its non-permissible cells.Our findings have not only identified chANXA2 as a novel biomarker for anti-ALV-J, but also demonstrated that cell lines with the expression of viral receptor-binding protein could be as efficient tools for isolating functional receptors to identify novel anti-viral targets.

View Article: PubMed Central - PubMed

Affiliation: 1] Ministry of Education Key Lab for Avian Preventive Medicine, Yangzhou University, No.12 East Wenhui Road, Yangzhou, Jiangsu. 225009, P. R. China [2] Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou. 225009, P. R. China.

ABSTRACT
The viral cell receptors and infection can be blocked by the expression of the viral receptor-binding protein. Thus, the viral cell receptor is an attractive target for anti-viral strategies, and the identification of viral cell receptor is critical for better understanding and controlling viral disease. As a model system for viral entry and anti-retroviral approaches, avian sarcoma/leukosis virus (ASLV, including the A-J ten subgroups) has been studied intensively and many milestone discoveries have been achieved based on work with ASLV. Here, we used a DF1 cell line expressed viral receptor-binding protein to efficiently identify chicken Annexin A2 (chANXA2) as a novel receptor for retrovirus ALV-J (avian leukosis virus subgroup J). Our data demonstrate that antibodies or siRNA to chANXA2 significantly inhibited ALV-J infection and replication, and over-expression of chANXA2 permitted the entry of ALV-J into its non-permissible cells. Our findings have not only identified chANXA2 as a novel biomarker for anti-ALV-J, but also demonstrated that cell lines with the expression of viral receptor-binding protein could be as efficient tools for isolating functional receptors to identify novel anti-viral targets.

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Related in: MedlinePlus

(Qin) siRNA against chANXA2 in DF1 cells DF1 cells were transfected with siRNA (50 pmol) against chANXA2 for 6 hr, and then the cells were infected with ALV-J at an MOI of 1 for 72 hr.(A), The siRNA effects on chANXA2 was detected with real-time PCR; (B), The infection/replication of ALV-J was analyzed with western blot. Lane 1, 2, 3 and 4, DF1 cells transfected with siRNA Mix (co-transfected with 535, 105 and 299), 535, 105 and 299 against chANXA2 respectively; lane5, DF1 cells transfected with control siRNA; lane6, DF1 cells with Mock.
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f3: (Qin) siRNA against chANXA2 in DF1 cells DF1 cells were transfected with siRNA (50 pmol) against chANXA2 for 6 hr, and then the cells were infected with ALV-J at an MOI of 1 for 72 hr.(A), The siRNA effects on chANXA2 was detected with real-time PCR; (B), The infection/replication of ALV-J was analyzed with western blot. Lane 1, 2, 3 and 4, DF1 cells transfected with siRNA Mix (co-transfected with 535, 105 and 299), 535, 105 and 299 against chANXA2 respectively; lane5, DF1 cells transfected with control siRNA; lane6, DF1 cells with Mock.

Mentions: To further evaluate whether the low expression of chANXA2 could confer resistance to ALV-J infection in its susceptible cells, we did siRNA against chANXA2 in DF1 cell, and then tested the effect on the ALV-J infection/replication. As described in Fig. 3A, real-time PCR showed that all four siRNA against chANXA2 tested could efficiently reduce chANXA2 mRNA level. And the western blot showed that all these four siRNA against chANXA2 tested could significantly decrease the infection/replication of ALV-J in DF1 cells (Fig. 3B).


Identification of novel viral receptors with cell line expressing viral receptor-binding protein.

Mei M, Ye J, Qin A, Wang L, Hu X, Qian K, Shao H - Sci Rep (2015)

(Qin) siRNA against chANXA2 in DF1 cells DF1 cells were transfected with siRNA (50 pmol) against chANXA2 for 6 hr, and then the cells were infected with ALV-J at an MOI of 1 for 72 hr.(A), The siRNA effects on chANXA2 was detected with real-time PCR; (B), The infection/replication of ALV-J was analyzed with western blot. Lane 1, 2, 3 and 4, DF1 cells transfected with siRNA Mix (co-transfected with 535, 105 and 299), 535, 105 and 299 against chANXA2 respectively; lane5, DF1 cells transfected with control siRNA; lane6, DF1 cells with Mock.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4300512&req=5

f3: (Qin) siRNA against chANXA2 in DF1 cells DF1 cells were transfected with siRNA (50 pmol) against chANXA2 for 6 hr, and then the cells were infected with ALV-J at an MOI of 1 for 72 hr.(A), The siRNA effects on chANXA2 was detected with real-time PCR; (B), The infection/replication of ALV-J was analyzed with western blot. Lane 1, 2, 3 and 4, DF1 cells transfected with siRNA Mix (co-transfected with 535, 105 and 299), 535, 105 and 299 against chANXA2 respectively; lane5, DF1 cells transfected with control siRNA; lane6, DF1 cells with Mock.
Mentions: To further evaluate whether the low expression of chANXA2 could confer resistance to ALV-J infection in its susceptible cells, we did siRNA against chANXA2 in DF1 cell, and then tested the effect on the ALV-J infection/replication. As described in Fig. 3A, real-time PCR showed that all four siRNA against chANXA2 tested could efficiently reduce chANXA2 mRNA level. And the western blot showed that all these four siRNA against chANXA2 tested could significantly decrease the infection/replication of ALV-J in DF1 cells (Fig. 3B).

Bottom Line: Thus, the viral cell receptor is an attractive target for anti-viral strategies, and the identification of viral cell receptor is critical for better understanding and controlling viral disease.Our data demonstrate that antibodies or siRNA to chANXA2 significantly inhibited ALV-J infection and replication, and over-expression of chANXA2 permitted the entry of ALV-J into its non-permissible cells.Our findings have not only identified chANXA2 as a novel biomarker for anti-ALV-J, but also demonstrated that cell lines with the expression of viral receptor-binding protein could be as efficient tools for isolating functional receptors to identify novel anti-viral targets.

View Article: PubMed Central - PubMed

Affiliation: 1] Ministry of Education Key Lab for Avian Preventive Medicine, Yangzhou University, No.12 East Wenhui Road, Yangzhou, Jiangsu. 225009, P. R. China [2] Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou. 225009, P. R. China.

ABSTRACT
The viral cell receptors and infection can be blocked by the expression of the viral receptor-binding protein. Thus, the viral cell receptor is an attractive target for anti-viral strategies, and the identification of viral cell receptor is critical for better understanding and controlling viral disease. As a model system for viral entry and anti-retroviral approaches, avian sarcoma/leukosis virus (ASLV, including the A-J ten subgroups) has been studied intensively and many milestone discoveries have been achieved based on work with ASLV. Here, we used a DF1 cell line expressed viral receptor-binding protein to efficiently identify chicken Annexin A2 (chANXA2) as a novel receptor for retrovirus ALV-J (avian leukosis virus subgroup J). Our data demonstrate that antibodies or siRNA to chANXA2 significantly inhibited ALV-J infection and replication, and over-expression of chANXA2 permitted the entry of ALV-J into its non-permissible cells. Our findings have not only identified chANXA2 as a novel biomarker for anti-ALV-J, but also demonstrated that cell lines with the expression of viral receptor-binding protein could be as efficient tools for isolating functional receptors to identify novel anti-viral targets.

Show MeSH
Related in: MedlinePlus