Th17 cell differentiation induced by cytopathogenic biotype BVDV-2 in bovine PBLCs

Abstract Background Bovine viral diarrhea virus (BVDV) is a major pathogen that causes bovine viral diarrhea/mucosal disease (BVD-MD), which has become a global infectious disease due to its wide spread and the lack of effective treatment. The process of BVDV infection is complex. Once infected, host immune cells are activated and modulated. As a major immune cell, peripheral blood lymphocyte cells (PBLCs) are the primary target of BVDV. In order to further understand the mechanism of BVDV- host interaction, the expression profiles of host lymphocytes mRNAs associated with BVDV infection were investigated by transcriptomic sequencing analysis. Results The transcriptomic sequencing analysis was performed on bovine PBLCs infected with CP BVDV-2 GS2018 after 12 h of infection. Gene expression profiling demonstrated that 1052 genes were differentially expressed in GS2018 infected PBLCs compared with the control group. Of these genes, 485 genes were up-regulated and 567 were down-regulated. The 19 differential expressed genes (DEGs) were selected for validation using quantitative real-time PCR and the results were consistent with the results of RNA-Seq. Gene ontology enrichment and KEGG pathway analysis showed that 1052 DEGs were significantly enriched in 16 pathways, including cytokine-cytokine receptor interaction, IL17, PI3K-Akt, MAPK and TNF signaling pathway. PPI network analysis showed that IL17A, IFN-γ and TNF-α interacted with various proteins and may play crucial roles in BVDV-2 infection. Of note, we confirmed that GS2018 induced Th17 cell differentiation in PBLCs and persistently increased the expression levels of IL17A. In turn, the replication of GS2018 was inhibited by IL17A. Conclusion In this study, the transcription changes of DEGs related to host immune responses in bovine PBLCs were caused by CP BVDV-2 infection. In particular, the effector molecules IL17A of Th17 cells were significantly up-regulated, which inhibited viral replication. These results will contribute to exploration and further understanding of the host immune response mechanism and interaction between host and BVDV-2.

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Th17 Cell Differentiation Induced by Cytopathogenic Biotype BVDV-2 in Bovine PBLCs

10.21203/rs.3.rs-422115/v1 ◽

2021 ◽

Author(s):

Yanping Li ◽

Tingli Liu ◽

Guoliang Chen ◽

Liqun Wang ◽

Aimin Guo ◽

...

Keyword(s):

Cell Differentiation ◽

Signaling Pathway ◽

Th17 Cell ◽

Bovine Viral Diarrhea ◽

Control Group ◽

Sequencing Analysis ◽

Diarrhea Virus ◽

Viral Diarrhea ◽

Th17 Cell Differentiation ◽

Transcriptomic Sequencing

Abstract Background: Bovine viral diarrhea virus (BVDV) is a major pathogen that causes bovine viral diarrhea/mucosal disease (BVD-MD), which has become a global infectious disease due to its wide spread and the lack of effective treatment. The process of BVDV infection is complex. Once infected, host immune cells are activated and modulated. As a major immune cell, peripheral blood lymphocyte cells (PBLCs) are the primary target of BVDV. In order to further understand the mechanism of BVDV- host interaction, the expression profiles of host lymphocytes mRNAs associated with BVDV infection were investigated by transcriptomic sequencing analysis. Results: The transcriptomic sequencing analysis was performed on bovine PBLCs infected with CP BVDV-2 GS2018 after 12 h of infection. Gene expression profiling demonstrated that 1,052 genes were differentially expressed in GS2018 infected PBLCs compared with the control group. Of these genes, 485 genes were up-regulated and 567 were down-regulated. 19 differential expressed genes (DEGs) were selected for validation using quantitative real-time PCR and the results were consistent with the results of RNA-Seq. Gene ontology enrichment and KEGG pathway analysis showed that 1052 DEGs were significantly enriched in 16 pathways, including cytokine-cytokine receptor interaction, IL17 signaling pathway, PI3K-Akt signaling pathway, MAPK signaling pathway and TNF signaling pathway. PPI network analysis showed that IL17A, IFN-γ and TNF-α interacted with various proteins and may play crucial roles in BVDV-2 infection. Of note, we confirmed that GS2018 induced Th17 cell differentiation in PBLCs and persistently increased the expression levels of IL17A. In turn, the replication of GS2018 was inhibited by IL17A. Conclusion: In this study, the transcription changes of DEGs related to host immune responses in bovine PBLCs were caused by CP BVDV-2 infection. In particular, the effector molecules IL17A of Th17 cells were significantly up-regulated, which inhibited viral replication. These results will contribute to exploration and further understanding of the host immune response mechanism and interaction between host and BVDV-2.

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Isolation and Genetic Analysis of Bovine Viral Diarrhea Virus from Infected Cattle in Indiana

Veterinary Medicine International ◽

10.4061/2011/925910 ◽

2011 ◽

Vol 2011 ◽

pp. 1-6 ◽

Cited By ~ 5

Author(s):

Roman M. Pogranichniy ◽

Megan E. Schnur ◽

Eran A. Raizman ◽

Duane A. Murphy ◽

Maria Negron ◽

...

Keyword(s):

Bovine Viral Diarrhea Virus ◽

Clinical Signs ◽

Bovine Viral Diarrhea ◽

Sequencing Analysis ◽

Diagnostic Laboratory ◽

Diarrhea Virus ◽

Viral Diarrhea ◽

Mucosal Disease ◽

Pcr Product ◽

Viral Diarrhea Virus

Species and biotype distribution was determined in 44 bovine viral diarrhea virus- (BVDV-) positive samples submitted to the Animal Disease Diagnostic Laboratory (ADDL) in Indiana during 2006–2008. BVDV RNA was detected in the 5′-untranslated region and Nproregion using reverse transcriptase PCR followed by sequencing analysis of the PCR product. Additionally, cases were classified into one of six categories according to history and/or lesions: acute symptomatic, hemorrhagic, respiratory distress, reproductive, persistent infection (PI), and mucosal disease (MD). Of 44 BVDV-positive samples, 33 were noncytopathic (ncp), 10 were cytopathic (cp), and one presented both ncp and cp biotypes. Sequencing analysis demonstrated that all samples belonged to BVDV-1a, BVDV-1b, or BVDV-2. The most common isolate was ncp BVDV-1b, (44%) followed by ncp BVDV-2a (24%). Among the six categories, respiratory clinical signs were the most common (36%) followed by PI (25%) and MD (16%).

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A β-Hairpin Motif in the Envelope Protein E2 Mediates Receptor Binding of Bovine Viral Diarrhea Virus

Viruses ◽

10.3390/v13061157 ◽

2021 ◽

Vol 13 (6) ◽

pp. 1157

Author(s):

Fernando Merwaiss ◽

María José Pascual ◽

María Trinidad Pomilio ◽

María Gabriela Lopez ◽

Oscar A. Taboga ◽

...

Keyword(s):

Bovine Viral Diarrhea Virus ◽

Envelope Protein ◽

Viral Population ◽

Cell Culture Supernatant ◽

Receptor Interaction ◽

Bovine Viral Diarrhea ◽

Wild Type ◽

Diarrhea Virus ◽

Viral Diarrhea ◽

Viral Diarrhea Virus

Pestivirus envelope protein E2 is crucial to virus infection and accomplishes virus-receptor interaction during entry. However, mapping of E2 residues mediating these interactions has remained unexplored. In this study, to investigate the structure-function relationship for a β-hairpin motif exposed to the solvent in the crystal structure of bovine viral diarrhea virus (BVDV) E2, we designed two amino acidic substitutions that result in a change of electrostatic potential. First, using wild type and mutant E2 expressed as soluble recombinant proteins, we found that the mutant protein had reduced binding to susceptible cells compared to wild type and diminished ability to inhibit BVDV infection, suggesting a lower affinity for BVDV receptors. We then analyzed the effect of β-hairpin mutations in the context of recombinant viral particles. Mutant viruses recovered from cell culture supernatant after transfection of recombinant RNA had almost completely inhibited ability to re-infect susceptible cells, indicating an impact of mutations on BVDV infectivity. Finally, sequential passaging of the mutant virus resulted in the selection of a viral population in which β-hairpin mutations reverted to the wild type sequence to restore infectivity. Taken together, our results show that this conserved region of the E2 protein is critical for the interaction with host cell receptors.

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A CRISPR/Cas9 Generated Bovine CD46-knockout Cell Line—A Tool to Elucidate the Adaptability of Bovine Viral Diarrhea Viruses (BVDV)

Viruses ◽

10.3390/v12080859 ◽

2020 ◽

Vol 12 (8) ◽

pp. 859 ◽

Cited By ~ 2

Author(s):

Kevin P. Szillat ◽

Susanne Koethe ◽

Kerstin Wernike ◽

Dirk Höper ◽

Martin Beer

Keyword(s):

Amino Acid ◽

Cell Line ◽

Receptor Interaction ◽

Bovine Viral Diarrhea ◽

Viral Glycoprotein ◽

Diarrhea Virus ◽

Viral Diarrhea ◽

Dependent Cell ◽

Viral Diarrhea Virus

Bovine viral diarrhea virus (BVDV) entry into a host cell is mediated by the interaction of the viral glycoprotein E2 with the cellular transmembrane CD46 receptor. In this study, we generated a stable Madin–Darby Bovine Kidney (MDBK) CD46-knockout cell line to study the ability of different pestivirus A and B species (BVDV-1 and -2) to escape CD46-dependent cell entry. Four different BVDV-1/2 isolates showed a clearly reduced infection rate after inoculation of the knockout cells. However, after further passaging starting from the remaining virus foci on the knockout cell line, all tested virus isolates were able to escape CD46-dependency and grew despite the lack of the entry receptor. Whole-genome sequencing of the escape-isolates suggests that the genetic basis for the observed shift in infectivity is an amino acid substitution of an uncharged (glycine/asparagine) for a charged amino acid (arginine/lysine) at position 479 in the ERNS in three of the four isolates tested. In the fourth isolate, the exchange of a cysteine at position 441 in the ERNS resulted in a loss of ERNS dimerization that is likely to influence viral cell-to-cell spread. In general, the CD46-knockout cell line is a useful tool to analyze the role of CD46 for pestivirus replication and the virus–receptor interaction.

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Immune Regulation of TNFAIP3 in Psoriasis through Its Association with Th1 and Th17 Cell Differentiation and p38 Activation

Journal of Immunology Research ◽

10.1155/2020/5980190 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Yanyun Jiang ◽

Wenming Wang ◽

Xiaofeng Zheng ◽

Hongzhong Jin

Keyword(s):

Cell Differentiation ◽

Mouse Model ◽

Mrna Expression ◽

Immune Regulation ◽

Th17 Cell ◽

Th17 Cells ◽

Serum Levels ◽

Control Group ◽

Expression Levels ◽

Th17 Cell Differentiation

Background. Psoriasis is an immune-mediated chronic inflammatory skin disorder in which the dysregulation of immune cells plays an important role in its development. Tumor necrosis factor- (TNF-) α antagonists affect the immune repertoire, while TNF-α-induced protein 3 (TNFAIP3) has a protective role against the deleterious effects of inflammation and participates in immune regulation. Objective. We investigated the immune regulation of TNFAIP3 in the pathogenesis of psoriasis and determined whether it is involved in the antipsoriatic effect of TNF-α antagonists. Methods. mRNA levels were evaluated in blood from patients with moderate-to-severe psoriasis. The effects of TNF-α antagonists were examined in a mouse imiquimod- (IMQ-) induced psoriasis-like dermatitis model. In the mouse model, TNFAIP3 mRNA expression was determined using RT-PCR. Serum levels of IL-17A, IL-23, IFN-γ, TNF-α, phosphorylated ERK1/2, p38, and JNK were measured using ELISA. The proportion of Th1 and Th17 cells in mouse spleens was analyzed using flow cytometry. Results. mRNA expression levels of TNFAIP3 in the blood were significantly lower in patients with moderate and severe psoriasis (mean±SD=0.44±0.25) compared with normal subjects (mean±SD=1.00±0.82) (P<0.01). In the mouse model, IMQ downregulated TNFAIP3 expression levels, which were increased after TNF-α antagonist treatment (P<0.05). Serum levels of Th17 cytokines (IL-17A and IL-23) and Th1 cytokines (IFN-γ and TNF-α) were significantly higher in the IMQ and IMQ/rat IgG1 groups compared with the control group, and the application of TNF-α antagonists significantly decreased the levels of inflammatory cytokines (P<0.01). Notably, phosphorylated p38 levels were increased in the IMQ and IMQ/rat IgG1 groups compared with the control group but were downregulated by treatment with TNF-α antagonists (P<0.05). Th1 and Th17 cells were significantly increased in the IMQ group compared with the control group (P<0.01). Conclusion. TNFAIP3 downregulation associated with Th1 and Th17 cell differentiation and p38 activation might contribute in part to the mechanism of immune dysfunction in psoriasis. TNF-α antagonists might partly exert their effects on psoriasis via this pathway.

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Actinobacteria from Termite Mounds Show Antiviral Activity against Bovine Viral Diarrhea Virus, a Surrogate Model for Hepatitis C Virus

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2015/745754 ◽

2015 ◽

Vol 2015 ◽

pp. 1-9 ◽

Cited By ~ 7

Author(s):

Marina Aiello Padilla ◽

Rodney Alexandre Ferreira Rodrigues ◽

Juliana Cristina Santiago Bastos ◽

Matheus Cavalheiro Martini ◽

Ana Caroline de Souza Barnabé ◽

...

Keyword(s):

Antiviral Activity ◽

Bovine Viral Diarrhea Virus ◽

Bovine Viral Diarrhea ◽

Rrna Gene ◽

Sequencing Analysis ◽

Termite Mounds ◽

Methanol Fraction ◽

Diarrhea Virus ◽

Viral Diarrhea ◽

Viral Diarrhea Virus

Extracts from termite-associated bacteria were evaluated forin vitroantiviral activity against bovine viral diarrhea virus (BVDV). Two bacterial strains were identified as active, with percentages of inhibition (IP) equal to 98%. Both strains were subjected to functional analysis via the addition of virus and extract at different time points in cell culture; the results showed that they were effective as posttreatments. Moreover, we performed MTT colorimetric assays to identify the CC50, IC50, and SI values of these strains, and strain CDPA27 was considered the most promising. In parallel, the isolates were identified asStreptomycesthrough 16S rRNA gene sequencing analysis. Specifically, CDPA27 was identified asS. chartreusis. The CDPA27 extract was fractionated on a C18-E SPE cartridge, and the fractions were reevaluated. A 100% methanol fraction was identified to contain the compound(s) responsible for antiviral activity, which had an SI of 262.41. GC-MS analysis showed that this activity was likely associated with the compound(s) that had a peak retention time of 5 min. Taken together, the results of the present study provide new information for antiviral research using natural sources, demonstrate the antiviral potential ofStreptomyces chartreusiscompounds isolated from termite mounds against BVDV, and lay the foundation for further studies on the treatment of HCV infection.

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