Deoxynivalenol Affects Proliferation and Expression of Activation-Related Molecules in Major Porcine T-Cell Subsets
Abstract
:1. Introduction
2. Results
2.1. Cell Viability and Lymphocyte Proliferation
2.2. Effect of DON on the Expression of the Activation Marker CD8α and the Costimulatory Molecules CD27 and CD28 within the Major T-cell Subsets
2.2.1. CD8α Expression in the Presence of Different DON Concentrations
2.2.2. CD28 Expression in the Presence of Different DON Concentrations
2.2.3. CD27 Expression in the Presence of Different DON Concentrations
2.3. Proliferation of CD27-Defined CD4+ and CD8+ T-Cell Subsets in the Presence of DON
2.4. Phenotypic Changes in CD8α and CD27 Expression in Proliferating Sorted and Unsorted Naïve CD4+ T Cells in the Presence of DON
3. Discussion
4. Materials and Methods
4.1. Animals and Cell Isolation
4.2. Proliferation Assays
4.3. Flow Cytometry Staining and Antibodies
4.4. Fluorescence-Activated Cell Sorting (FACS) and Antibodies
4.5. Proliferation Assay and In Vitro Cultivation of Sorted T Cells
4.6. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Antigen | Clone | Isotype | Fluorochrome | Labeling Strategy | Source of Primary Ab |
---|---|---|---|---|---|
Analysis of co-Stimulatory Molecules | |||||
CD3 | BB23-8E6-8C8 | IgG2a | PerCP-Cy5.5 | Directly conjugated | BD Biosciences |
CD4 | 74-12-4 | IgG2b | FITC | Directly conjugated | BD Biosciences |
CD8α | 11/295/33 | IgG2a | Alexa647 | Directly conjugated | In house |
TCR-γδ | PPT16 | IgG2b | BV605 | Biotin-Streptavidin 3 | In house |
CD28 1 | 3D11 2 | IgG1 | PE | Secondary antibody 4 | In house 2 |
CD27 1 | b30c7 | IgG1 | PE | Secondary antibody 4 | In house |
Analysis of Perforin, CD27 and CD8α Expression | |||||
CD27 | b30c7 | IgG1 | Alexa647 | Secondary antibody 5 | In house |
CD4 | 74-12-4 | IgG2b | FITC | Secondary antibody 6 | In house |
CD8α 1 | 11/295/33 | IgG2α | PE-Cy7 | Secondary antibody 7 | In house |
CD8β 1 | PG164a | IgG2a | PE-Cy7 | Secondary antibody 7 | In house |
Perforin | δ-G9 | IgG2b | PerCP-eFluor710 | Directly conjugated | eBioscience |
Antigen | Clone | Isotype | Fluorochrome | Labeling Strategy | Source of Primary Ab |
---|---|---|---|---|---|
Staining for Cell Sorting | |||||
CD4 | 74-12-4 | IgG2b | FITC | Directly conjugated | BD Biosciences |
CD27 | b30c7 | IgG1 | PE | Secondary antibody 1 | In house |
CD8α | 11/295/33 | IgG2a | Alexa647 | Secondary antibody 2 | In house |
CD172a | 77-22-15A | IgG2b | eFluor450 | Biotin-Streptavidin 3 | In house |
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Vatzia, E.; Pierron, A.; Saalmüller, A.; Mayer, E.; Gerner, W. Deoxynivalenol Affects Proliferation and Expression of Activation-Related Molecules in Major Porcine T-Cell Subsets. Toxins 2019, 11, 644. https://doi.org/10.3390/toxins11110644
Vatzia E, Pierron A, Saalmüller A, Mayer E, Gerner W. Deoxynivalenol Affects Proliferation and Expression of Activation-Related Molecules in Major Porcine T-Cell Subsets. Toxins. 2019; 11(11):644. https://doi.org/10.3390/toxins11110644
Chicago/Turabian StyleVatzia, Eleni, Alix Pierron, Armin Saalmüller, Elisabeth Mayer, and Wilhelm Gerner. 2019. "Deoxynivalenol Affects Proliferation and Expression of Activation-Related Molecules in Major Porcine T-Cell Subsets" Toxins 11, no. 11: 644. https://doi.org/10.3390/toxins11110644