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- Title
Activation of NF-κB pathway is involved in secretion of IL-4 and IL-12 in PCV2-infected piglet lymphocytes in vitro.
- Authors
DUAN Dianning; GUO Hua; LI Xiaoling; CHEN Mengmeng; ZHANG Yaqun; HAN Junyuan; ZHANG Shuxia
- Abstract
Eight post-weaning piglets, which were free of porcine circorirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV), were chosen as the experimental animals. The suspensions of the single cells isolated from the spleens of these piglets were divided into four groups as κfollows: control group, nuclear factor kappa B( NF-B) inhibitor group( inhibitor group), PCV2 group,and NF-κB inhibitor-PCV2 group (inhibitor-PCV2 group). After 24 h incubation in vitro,the lymphocytes and the resulting supernatants were harvested respectively. The concentrations of interleukin 12 ( IL-12) and IL-4 secreted in the supernatants were measured by ELISA. The observation of the PCV2-infected cells and the nuclear translocation of NF-κB were conducted using indirect immunofluorescence assay. The protein concentrations of myeloiddifferentiation factor 88 ( MyD88), phosphorylated IκBα(p-IκBα),NF-κB/ p65 in cytoplasm and NF-κB/ p65 in nucleus were detected by Western blot. The NF-κB- DNA binding activity in nucleus was detected using electrophoretic mobility shift assay( EMSA). Results as follows: The IL-4 concentration in the PCV2 group was significantly lower than that in the control group as well as in the inhibitor-PCV2 group( P< 0.05), while the IL-4 concentration in the control group was similar to that in the inhibitor-PCV2 group. The IL-12 concentration in the PCV2 group was notably higher than that in the control group(P<0. 05)as well as in the inhibitor-PCV2 group(P<0. 01). After PCV2 infection the lymphocytes,the PCV2 antigens were mostly observed in the lymphocytes cytoplasm. The protein concentration of MyD88 expressed in the lymphocyte cytoplasm in the PCV2 group was markedly higher compared to that in control group(P<0. 01), but was similar to that in the inhibitor-κPCV2 group. For the PCV2 group,the protein concentrations of p-κI?B in the lymphocyte cytoplasm and NF-κB/ p65 in the nucleus,and the NF-κB-DNA binding activity were higher than those in the control group;in contrast,those in the inhibitor group as well as in the inhibitor-PCV2 group were dramatically lower than those in the control group. Conclusions:PCV2 may infect piglet lymphocytes in vitro and mediated the activation of MyD88-NF-κB signaling pathway,which is involved in secretion of IL-4 and IL-12 in the cells.
- Publication
Journal of Nanjing Agricultural University / Nanjuing Nongye Daxue Xuebao, 2014, Vol 37, Issue 3, p93
- ISSN
1000-2030
- Publication type
Article
- DOI
10.7685/j. issn.1000-2030.2014.03.010