Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Feb 24;9(3):101.
doi: 10.3390/vetsci9030101.

In Vitro Analysis of TGF-β Signaling Modulation of Porcine Alveolar Macrophages in Porcine Circovirus Type 2b Infection

Shunli Yang  1 , Muhammad Umar Zafar Khan  1   2 , Baohong Liu  1 , Muhammad Humza  3 , Shuanghui Yin  1 , Jianping Cai  1   4
Affiliations

In Vitro Analysis of TGF-β Signaling Modulation of Porcine Alveolar Macrophages in Porcine Circovirus Type 2b Infection

Shunli Yang et al. Vet Sci. .

Abstract

Porcine circovirus 2 (PCV2) has been recognized as an immunosuppressive pathogen. However, the crosstalk between this virus and its host cells in related signaling pathways remains poorly understood. In this study, the expression profiles of 84 genes involved in transforming growth factor-beta (TGF-β) signaling pathway were probed in PCV2b-infected primary porcine alveolar macrophages (PAMs) by using an RT2 profiler PCR array system. The protein expression levels of cytokines involved in the TGF-β signaling pathway were determined with a RayBiotech fluorescent Quantibody® porcine cytokine array system. Results showed that 48, 30, and 42 genes were differentially expressed at 1, 24, and 48 h after infection, respectively. A large number of genes analyzed by a co-expression network and implicated in transcriptional regulation and apoptosis were differentially expressed in PCV2b-infected PAMs. Among these genes, TGF-β, interleukin-10, CCAAT/enhancer-binding protein beta (C/EBPB), growth arrest, and DNA-damage-inducible 45 beta (GADD45B), and BCL2 were upregulated. By contrast, SMAD family member 1 (smad1) and smad3 were downregulated. These results suggested that the TGF-β signaling pathway was repressed in PAMs at the early onset of PCV2 infection. The inhibited apoptosis was indicated by the upregulated C/EBPB, GADD45B, and BCL2, and by the downregulated smad1 and smad3, which possibly increased the duration of PCV2 replication-permissive conditions and caused a persistent infection. Our study may provide insights into the underlying antiviral functional changes in the immune system of PCV2-infected pigs.

Keywords: mRNA; porcine circovirus 2; signaling pathway analysis; transforming growth factor-beta.

PubMed Disclaimer

Conflict of interest statement

None of the authors of this paper has a financial or personal relationship with other people or organizations that could inappropriately influence the content of this paper.

Figures

Figure 1
Figure 1
PCV2 infection in PAMs. PCV2 in PAMs post-infection was detected by IFA based on the ODs-psdAb probe (red), the nucleus was stained with DAPI (blue).
Figure 2
Figure 2
Gene expression patterns. Sixty-three target genes involved in the TGF-β signaling pathway were differentially expressed and were significantly different between PCV2-infected and control PAMs (A). Differentially expressed genes at each time point (B). Green: negative fold change level. Red: positive fold change level. Fold change was obtained by comparing the relative mRNA expression of target genes in PCV2-infected and uninfected control PAMs. Results are representative of three independent experiments and are presented as mean ± SE. ** represents p <0.01, * represents p < 0.05.
Figure 3
Figure 3
Expression of TGF-β at the protein level. TGF-β in the culture supernatants of control and PCV2-infected PAMs were measured by using porcine cytokine array kits. Data are expressed as pg/mL of three independent experiments. * represents p < 0.05.
Figure 4
Figure 4
Co-expression network. A co-expression network from the expression profiles of 84 TGF-β signaling pathway-related genes was constructed. Interactive modules (A), Clustering coefficients (B).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Meng X.J. Spread like a wildfire--the omnipresence of porcine circovirus type 2 (PCV2) and its ever-expanding association with diseases in pigs. Virus Res. 2012;164:1–3. doi: 10.1016/j.virusres.2011.12.005. - DOI - PubMed
    1. Ge X., Wang F., Guo X., Yang H. Porcine circovirus type 2 and its associated diseases in China. Virus Res. 2012;164:100–106. doi: 10.1016/j.virusres.2011.10.005. - DOI - PubMed
    1. Segales J., Rose C., II, Domingo M. Pathological findings associated with naturally acquired porcine circovirus type 2 associated disease. Vet. Microbiol. 2004;98:137–149. doi: 10.1016/j.vetmic.2003.10.006. - DOI - PubMed
    1. Darwich L., Segales J., Domingo M., Mateu E. Changes in CD4(+), CD8(+), CD4(+) CD8(+), and immunoglobulin M-positive peripheral blood mononuclear cells of postweaning multisystemic wasting syndrome-affected pigs and age-matched uninfected wasted and healthy pigs correlate with lesions and porcine circovirus type 2 load in lymphoid tissues. Clin. Diagn. Lab. Immunol. 2002;9:236–242. - PMC - PubMed
    1. Segales J., Domingo M., Chianini F., Majo N., Dominguez J., Darwich L., Mateu E. Immunosuppression in postweaning multisystemic wasting syndrome affected pigs. Vet. Microbiol. 2004;98:151–158. doi: 10.1016/j.vetmic.2003.10.007. - DOI - PubMed

LinkOut - more resources