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The 15N and 46R Residues of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein Enhance Regulatory T Lymphocytes Proliferation.

Fan B, Liu X, Bai J, Li Y, Zhang Q, Jiang P - PLoS ONE (2015)

Bottom Line: Porcine reproductive and respiratory syndrome virus (PRRSV) negatively modulates host immune responses, resulting in persistent infection and immunosuppression.By using reverse genetic methods, it was firstly found that the 15N and 46R residues in PRRSV N protein were critical for induction of Tregs proliferation.The phenotype of induced Tregs closely resembled that of transforming-growth-factor-β-secreting T helper 3 Tregs in swine.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Animal Diseases Diagnostic and Immunology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.

ABSTRACT
Porcine reproductive and respiratory syndrome virus (PRRSV) negatively modulates host immune responses, resulting in persistent infection and immunosuppression. PRRSV infection increases the number of PRRSV-specific regulatory T lymphocytes (Tregs) in infected pigs. However, the target antigens for Tregs proliferation in PRRSV infection have not been fully understood. In this study, we demonstrated that the highly pathogenic PRRSV (HP-PRRSV) induced more CD4+CD25+Foxp3+ Tregs than classical PRRSV (C-PRRSV) strain. Of the recombinant GP5, M and N proteins of HP-PRRSV expressed in baculovirus expression systems, only N protein induced Tregs proliferation. The Tregs assays showed that three amino-acid regions, 15-21, 42-48 and 88-94, in N protein played an important role in induction of Tregs proliferation with synthetic peptides covering the whole length of N protein. By using reverse genetic methods, it was firstly found that the 15N and 46R residues in PRRSV N protein were critical for induction of Tregs proliferation. The phenotype of induced Tregs closely resembled that of transforming-growth-factor-β-secreting T helper 3 Tregs in swine. These data should be useful for understanding the mechanism of immunity to PRRSV and development of infection control strategies in the future.

No MeSH data available.


Related in: MedlinePlus

Role of baculovirus-expressed proteins GP5, M and N of HP-PRRSV in induction of Tregs proliferation.(A) Purified recombinant GP5, M and N protein expressed in baculovirus. (B) Percentage of Foxp3+ cells in the gated CD4+ CD25+ subpopulations of GP5, M and N. (C) Concentrations of TGF-β in supernatants of 3-day co-cultures. (D) Concentrations of IL-10 in supernatants of 3-day co-cultures. Data came from three independent experiments. Data analysis was done using one-way ANOVA and significant differences are shown (*P<0.05, **P<0.01 and ***P<0.001).
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pone.0138772.g003: Role of baculovirus-expressed proteins GP5, M and N of HP-PRRSV in induction of Tregs proliferation.(A) Purified recombinant GP5, M and N protein expressed in baculovirus. (B) Percentage of Foxp3+ cells in the gated CD4+ CD25+ subpopulations of GP5, M and N. (C) Concentrations of TGF-β in supernatants of 3-day co-cultures. (D) Concentrations of IL-10 in supernatants of 3-day co-cultures. Data came from three independent experiments. Data analysis was done using one-way ANOVA and significant differences are shown (*P<0.05, **P<0.01 and ***P<0.001).

Mentions: To examine the effect of PRRSV structural proteins on induction of Tregs, the GP5, M and N proteins of HP-PRRSV were expressed in baculovirus expression systems and confirmed by western blotting with His antibody (Fig 3A). Recombinant GP5, M and N proteins were purified and used to examine their effects on Tregs induction. Only N protein significantly induced proliferation of Tregs (Fig 3B). The concentrations of TGF-β and IL-10 in the supernatants were quantified with ELISA. GP5 and N proteins significantly increased expression of IL-10 (P<0.05), but only N protein significantly increased TGF-β expression (P<0.05) (Fig 3C and 3D).


The 15N and 46R Residues of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein Enhance Regulatory T Lymphocytes Proliferation.

Fan B, Liu X, Bai J, Li Y, Zhang Q, Jiang P - PLoS ONE (2015)

Role of baculovirus-expressed proteins GP5, M and N of HP-PRRSV in induction of Tregs proliferation.(A) Purified recombinant GP5, M and N protein expressed in baculovirus. (B) Percentage of Foxp3+ cells in the gated CD4+ CD25+ subpopulations of GP5, M and N. (C) Concentrations of TGF-β in supernatants of 3-day co-cultures. (D) Concentrations of IL-10 in supernatants of 3-day co-cultures. Data came from three independent experiments. Data analysis was done using one-way ANOVA and significant differences are shown (*P<0.05, **P<0.01 and ***P<0.001).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4580451&req=5

pone.0138772.g003: Role of baculovirus-expressed proteins GP5, M and N of HP-PRRSV in induction of Tregs proliferation.(A) Purified recombinant GP5, M and N protein expressed in baculovirus. (B) Percentage of Foxp3+ cells in the gated CD4+ CD25+ subpopulations of GP5, M and N. (C) Concentrations of TGF-β in supernatants of 3-day co-cultures. (D) Concentrations of IL-10 in supernatants of 3-day co-cultures. Data came from three independent experiments. Data analysis was done using one-way ANOVA and significant differences are shown (*P<0.05, **P<0.01 and ***P<0.001).
Mentions: To examine the effect of PRRSV structural proteins on induction of Tregs, the GP5, M and N proteins of HP-PRRSV were expressed in baculovirus expression systems and confirmed by western blotting with His antibody (Fig 3A). Recombinant GP5, M and N proteins were purified and used to examine their effects on Tregs induction. Only N protein significantly induced proliferation of Tregs (Fig 3B). The concentrations of TGF-β and IL-10 in the supernatants were quantified with ELISA. GP5 and N proteins significantly increased expression of IL-10 (P<0.05), but only N protein significantly increased TGF-β expression (P<0.05) (Fig 3C and 3D).

Bottom Line: Porcine reproductive and respiratory syndrome virus (PRRSV) negatively modulates host immune responses, resulting in persistent infection and immunosuppression.By using reverse genetic methods, it was firstly found that the 15N and 46R residues in PRRSV N protein were critical for induction of Tregs proliferation.The phenotype of induced Tregs closely resembled that of transforming-growth-factor-β-secreting T helper 3 Tregs in swine.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Animal Diseases Diagnostic and Immunology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.

ABSTRACT
Porcine reproductive and respiratory syndrome virus (PRRSV) negatively modulates host immune responses, resulting in persistent infection and immunosuppression. PRRSV infection increases the number of PRRSV-specific regulatory T lymphocytes (Tregs) in infected pigs. However, the target antigens for Tregs proliferation in PRRSV infection have not been fully understood. In this study, we demonstrated that the highly pathogenic PRRSV (HP-PRRSV) induced more CD4+CD25+Foxp3+ Tregs than classical PRRSV (C-PRRSV) strain. Of the recombinant GP5, M and N proteins of HP-PRRSV expressed in baculovirus expression systems, only N protein induced Tregs proliferation. The Tregs assays showed that three amino-acid regions, 15-21, 42-48 and 88-94, in N protein played an important role in induction of Tregs proliferation with synthetic peptides covering the whole length of N protein. By using reverse genetic methods, it was firstly found that the 15N and 46R residues in PRRSV N protein were critical for induction of Tregs proliferation. The phenotype of induced Tregs closely resembled that of transforming-growth-factor-β-secreting T helper 3 Tregs in swine. These data should be useful for understanding the mechanism of immunity to PRRSV and development of infection control strategies in the future.

No MeSH data available.


Related in: MedlinePlus