Proteomic Characterization of PAMs with PRRSV-ADE Infection

Xu, Pengli; Li, Wen; Zhao, Shijie; Cui, Zhiying; Chen, Yu; Zhang, Yina; Chen, Jing; Xia, Pingan

doi:10.3390/v15010036

Cited by 5 publications

(5 citation statements)

References 45 publications

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“…All three tested negative for both PCV2 and PRRSV. PAMs were collected by bronchoalveolar lavage as previously described [ 23 ]. Before use, the cells were confirmed as negative for PCV2, PRRSV, pseudorabies virus (PRV) and classical swine fever virus (CSFV) by PCR.…”

Section: Methodsmentioning

confidence: 99%

Immune Molecules’ mRNA Expression in Porcine Alveolar Macrophages Co-Infected with Porcine Reproductive and Respiratory Syndrome Virus and Porcine Circovirus Type 2

Cui

Zhou

Hu³

et al. 2023

Viruses

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Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus 2 (PCV2) are economically important pathogens in swine, and pigs with dual infections of PCV2 and PRRSV consistently have more severe clinical symptoms and interstitial pneumonia. However, the synergistic pathogenesis mechanism induced by PRRSV and PCV2 co-infection has not yet been illuminated. Therefore, the aim of this study was to characterize the kinetic changes of immune regulatory molecules, inflammatory factors and immune checkpoint molecules in porcine alveolar macrophages (PAMs) in individuals infected or co-infected with PRRSV and/or PCV2. The experiment was divided into six groups: a negative control group (mock, no infected virus), a group infected with PCV2 alone (PCV2), a group infected with PRRSV alone (PRRSV), a PCV2–PRRSV co-infected group (PCV2–PRRSV inoculated with PCV2, followed by PRRSV 12 h later), a PRRSV–PCV2 co-infected group (PRRSV–PCV2 inoculated with PRRSV, followed by PCV2 12 h later) and a PCV2 + PRRSV co-infected group (PCV2 + PRRSV, inoculated with PCV2 and PRRSV at the same time). Then, PAM samples from the different infection groups and the mock group were collected at 6, 12, 24, 36 and 48 h post-infection (hpi) to detect the viral loads of PCV2 and PRRSV and the relative quantification of immune regulatory molecules, inflammatory factors and immune checkpoint molecules. The results indicated that PCV2 and PRRSV co-infection, regardless of the order of infection, had no effect on promoting PCV2 replication, while PRRSV and PCV2 co-infection was able to promote PRRSV replication. The immune regulatory molecules (IFN-α and IFN-γ) were significantly down-regulated, while inflammatory factors (TNF-α, IL-1β, IL-10 and TGF-β) and immune checkpoint molecules (PD-1, LAG-3, CTLA-4 and TIM-3) were significantly up-regulated in the PRRSV and PCV2 co-infection groups, especially in PAMs with PCV2 inoculation first followed by PRRSV. The dynamic changes in the aforementioned immune molecules were associated with a high viral load, immunosuppression and cell exhaustion, which may explain, at least partially, the underlying mechanism of the enhanced pulmonary lesions by dual infection with PCV2 and PRRSV in PAMs.

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Section: Methodsmentioning

confidence: 99%

Immune Molecules’ mRNA Expression in Porcine Alveolar Macrophages Co-Infected with Porcine Reproductive and Respiratory Syndrome Virus and Porcine Circovirus Type 2

Cui

Zhou

Hu³

et al. 2023

Viruses

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“…MHC-I and MHC-II expressions were suppressed in PRRSV-infected APCs ( 6 , 56 , 63 ). PRRSV-2 was reported to reduce STAT1, STAT2, and STAT6 expressions in MARC-145 cells and PAMs ( 64 , 65 ). HP-PRRSV was demonstrated to inhibit TNFα in PAMs.…”

Section: Discussionmentioning

confidence: 99%

Type I and II interferons, transcription factors and major histocompatibility complexes were enhanced by knocking down the PRRSV-induced transforming growth factor beta in monocytes co-cultured with peripheral blood lymphocytes

Fabros,

Charerntantanakul

2024

Front. Immunol.

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The innate and adaptive immune responses elicited by porcine reproductive and respiratory syndrome virus (PRRSV) infection are known to be poor. This study investigates the impact of PRRSV-induced transforming growth factor beta 1 (TGFβ1) on the expressions of type I and II interferons (IFNs), transcription factors, major histocompatibility complexes (MHC), anti-inflammatory and pro-inflammatory cytokines in PRRSV-infected co-cultures of monocytes and peripheral blood lymphocytes (PBL). Phosphorothioate-modified antisense oligodeoxynucleotide (AS ODN) specific to the AUG region of porcine TGFβ1 mRNA was synthesized and successfully knocked down TGFβ1 mRNA expression and protein translation. Monocytes transfected with TGFβAS1 ODN, then simultaneously co-cultured with PBL and inoculated with either classical PRRSV-2 (cPRRSV-2) or highly pathogenic PRRSV-2 (HP-PRRSV-2) showed a significant reduction in TGFβ1 mRNA expression and a significant increase in the mRNA expressions of IFNα, IFNγ, MHC-I, MHC-II, signal transducer and activator of transcription 1 (STAT1), and STAT2. Additionally, transfection of TGFβAS1 ODN in the monocyte and PBL co-culture inoculated with cPRRSV-2 significantly increased the mRNA expression of interleukin-12p40 (IL-12p40). PRRSV-2 RNA copy numbers were significantly reduced in monocytes and PBL co-culture transfected with TGFβAS1 ODN compared to the untransfected control. The yields of PRRSV-2 RNA copy numbers in PRRSV-2-inoculated monocytes and PBL co-culture were sustained and reduced by porcine TGFβ1 (rTGFβ1) and recombinant porcine IFNα (rIFNα), respectively. These findings highlight the strategy employed by PRRSV to suppress the innate immune response through the induction of TGFβ expression. The inclusion of TGFβ as a parameter for future PRRSV vaccine and vaccine adjuvant candidates is recommended.

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“…To explore the proteomic characterization of PRRSV infection, PAMs were infected with PRRSV at an MOI of 1 for 24 h, the cells of Mock group and PRRSV group were dissolved in lysis solution with SDT buffer (4% ( w / v ) SDS, 100 mM Tris-HCl pH 7.6, 0.1 M DTT) to extract the protein (n = 3). The TMT-based quantitative proteomic were performed at Applied Protein Technology Biotechnology Co., Ltd. (Shanghai, China) [ 4 ].…”

Section: Methodsmentioning

confidence: 99%

“…Porcine reproductive and respiratory syndrome (PRRS) is a viral disease caused by PRRS virus (PRRSV) with serious reproductive disorders in sows and respiratory symptoms in piglets, leading to significant economic losses in the pig industry worldwide. Genetic diversity, immunosuppression, and the antibody-dependent enhancement (ADE) effect of PRRSV pose additional challenges for PRRS prevention and control [ 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 ]. Although vaccination is the most effective method to prevent PRRSV infection, the efficacy of PRRSV vaccines remains unsatisfactory [ 9 , 10 , 11 ].…”

Section: Introductionmentioning

confidence: 99%

Mass Spectrometry-Based Proteomic Analysis of Potential Host Proteins Interacting with GP5 in PRRSV-Infected PAMs

Li,

Wang,

Zhang

et al. 2024

IJMS

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Porcine reproductive and respiratory syndrome virus (PRRSV) is a typical immunosuppressive virus causing a large economic impact on the swine industry. The structural protein GP5 of PRRSV plays a pivotal role in its pathogenicity and immune evasion. Virus–host interactions play a crucial part in viral replication and immune escape. Therefore, understanding the interactions between GP5 and host proteins are significant for porcine reproductive and respiratory syndrome (PRRS) control. However, the interaction network between GP5 and host proteins in primary porcine alveolar macrophages (PAMs) has not been reported. In this study, 709 GP5-interacting host proteins were identified in primary PAMs by immunoprecipitation coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Bioinformatics analysis revealed that these proteins were involved in multiple cellular processes, such as translation, protein transport, and protein stabilization. Subsequently, immunoprecipitation and immunofluorescence assay confirmed that GP5 could interact with antigen processing and presentation pathways related proteins. Finally, we found that GP5 may be a key protein that inhibits the antigen processing and presentation pathway during PRRSV infection. The novel host proteins identified in this study will be the candidates for studying the biological functions of GP5, which will provide new insights into PRRS prevention and vaccine development.

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Proteomic Characterization of PAMs with PRRSV-ADE Infection

Cited by 5 publications

References 45 publications

Immune Molecules’ mRNA Expression in Porcine Alveolar Macrophages Co-Infected with Porcine Reproductive and Respiratory Syndrome Virus and Porcine Circovirus Type 2

Immune Molecules’ mRNA Expression in Porcine Alveolar Macrophages Co-Infected with Porcine Reproductive and Respiratory Syndrome Virus and Porcine Circovirus Type 2

Type I and II interferons, transcription factors and major histocompatibility complexes were enhanced by knocking down the PRRSV-induced transforming growth factor beta in monocytes co-cultured with peripheral blood lymphocytes

Mass Spectrometry-Based Proteomic Analysis of Potential Host Proteins Interacting with GP5 in PRRSV-Infected PAMs

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