Journal: mBio

Article Title: PCV2 infection induces the differentiation of Treg cells via the TGF-β/Smad3 pathway

doi: 10.1128/mbio.01366-25

Figure Lengend Snippet: ( A ) CD4 + T cell immune balance detection. Th1 and Th2 cells were detected simultaneously by flow cytometry. Th1 cells were labeled with CD4 + IFN-γ + (PerCP-CyTM5.5 mouse anti-pig CD4α, Alexa Fluor 647 mouse anti-pig IFN-γ), and Th2 cells were labeled with CD4 + GATA3 + (PerCP-CyTM5.5 mouse anti-pig CD4α, Gata-3 monoclonal antibody [TWAJ], PE). ( B ) Th17 cells were detected by flow cytometry. They were labeled with CD4 + IL-17A + (PerCP-CyTM5.5 mouse anti-pig CD4α, PE mouse anti-human IL-17A). ( C ) Treg cells were detected by flow cytometry. They were labeled with CD4 + CD25 + Foxp3 + (PerCP-CyTM5.5 mouse anti-pig CD4α, Alexa Fluor 647 mouse anti-pig CD25, FOXP3 monoclonal antibody (FJK-16s), PE). Dynamic changes in Th1 ( D ) and Th2 ( E ) cell frequencies were quantitatively analyzed at various time points following PCV2 infection. ( F ) Th1/Th2 immune balance analysis. The frequencies of Th17 ( G ) and Treg ( H ) cells in piglets were measured at different time points during PCV2 infection. ( I ) Th17/Treg immune balance analysis.

Article Snippet: Alexa Flour 647 mouse anti-pig CD25 (MCA1736A647) was purchased from Bio-Rad.

Techniques: Flow Cytometry, Labeling, Infection

Journal: Advanced healthcare materials

Article Title: Biocompatibility of Water-Dispersible Pristine Graphene and Graphene Oxide Using a Close-to-Human Animal Model: A Pilot Study on Swine.

doi: 10.1002/adhm.202401783

Figure Lengend Snippet: Figure 6. GR or GO administration did not affect cytokine levels or early activation marker (CD25) expression on PBMCs. Pigs were injected intra- peritoneal with 15 mg of GR or GO alongside controls. Pre-injection (0) and after 1, 7, 14, and 21 days, EDTA blood was collected, PBMCs were purified, and plasma was collected. A) Plasma levels of IL-1𝛽, IL-2, IL-4, IL-6, IL-10, and IL-12 were monitored over time using a multiplex ELISA. Data from three controls or four treated different pigs are presented as mean + SD. B) CD25 (activation marker) expression on monocytes, B cells, NK cells, and T cells,

Article Snippet: The expression of CD25 (activation marker) was assessed using first mouse anti-pig CD25 (K231.3B2, Bio-rad) and subsequent staining with BV421 rat anti-mouse IgG1 (A85-1, BD Horizon).

Techniques: Activation Assay, Marker, Expressing, Injection, Clinical Proteomics, Multiplex Assay, Enzyme-linked Immunosorbent Assay

Journal: Advanced Healthcare Materials

Article Title: Biocompatibility of Water‐Dispersible Pristine Graphene and Graphene Oxide Using a Close‐to‐Human Animal Model: A Pilot Study on Swine

doi: 10.1002/adhm.202401783

Figure Lengend Snippet: GR or GO administration did not affect cytokine levels or early activation marker (CD25) expression on PBMCs. Pigs were injected intra‐peritoneal with 15 mg of GR or GO alongside controls. Pre‐injection (0) and after 1, 7, 14, and 21 days, EDTA blood was collected, PBMCs were purified, and plasma was collected. A) Plasma levels of IL‐1β, IL‐2, IL‐4, IL‐6, IL‐10, and IL‐12 were monitored over time using a multiplex ELISA. Data from three controls or four treated different pigs are presented as mean + SD. B) CD25 (activation marker) expression on monocytes, B cells, NK cells, and T cells, then subdivided into cytotoxic (CD8 high CD4 − ), memory T helper (CD8 + CD4 + ), naïve T helper (CD8 − CD4 + ) was determined using flow cytometry. Data from three (controls) or five (treated) different pigs are presented as mean + SD. C) Representative dot plots are displayed. At each time post‐injection, values of cytokines or activation markers were compared using a one‐way ANOVA followed by Bonferroni's multiple comparison test.

Article Snippet: The expression of CD25 (activation marker) was assessed using first mouse anti‐pig CD25 (K231.3B2, Bio‐rad) and subsequent staining with BV421 rat anti‐mouse IgG1 (A85‐1, BD Horizon).

Techniques: Activation Assay, Marker, Expressing, Injection, Purification, Clinical Proteomics, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Comparison

Journal: Frontiers in Veterinary Science

Article Title: Investigation of activation-induced markers (AIM) in porcine T cells by flow cytometry

doi: 10.3389/fvets.2024.1390486

Figure Lengend Snippet: Expression of AIMs in blood-derived CD3 + T cells after stimulation with PMA/ionomycin. (A) Representative FCM plots depicting expression of CD25, CD69, CD40L and TNF-α in CD3 + T cells when PBMC samples were unstimulated (Medium, top row) or stimulated with PMA/ionomycin (PMA, bottom row) for 18 h. Surface staining was performed for antibodies against CD3, CD4, CD8α, CD25 and CD69. Intracellular staining was performed for CD40L and TNF-α. Gates shown are representative of gating for total CD25 + , total CD69 + , total CD40L + and total TNF-α + T cells applied to PMA-stimulated samples and used in Boolean gating to create doughnut charts. (B) Doughnut charts of AIM phenotypes in PMA-stimulated samples generated by Boolean gating. Each doughnut represents the PBMC sample of one pig. Different phenotypes are indicated by different colors with all AIM phenotypes co-expressing TNF-α summarized in gray. CD25 − CD40L − CD69 − TNF-α − T cells are not shown. (C) Live CD3 + T cells from unstimulated (Medium), SEB-stimulated and PMA-stimulated cultures were clustered using the t-SNE algorithm with generated clusters shown in a colored overlay (left side). Relative expression levels of CD69, CD25 and CD40L within clusters (right side) are colored from high (red) to low (blue).

Article Snippet: Cells were surface-stained with primary monoclonal antibodies (mAbs) directed against CD3 (PerCP-Cy5.5-conjugated, mouse IgG2a anti-pig, clone: BB23-8E6-8C8, BD Biosciences), CD4 (FITC-conjugated, mouse IgG1 anti-pig, clone: b38c6c7, Bio-Rad Laboratories Ltd., Hercules, CA, United States), CD8α (biotinylated, mouse IgG2a anti-pig, clone: 76–2-11, Southern Biotech, Birmingham, AL, United States), CD25 (AlexaFluor 647-conjugated, mouse IgG1 anti-pig, clone: K231.3B2, Bio-Rad) and CD69 [unconjugated, mouse IgG2b anti-pig, clone: 01-14-22-51 ( )].

Techniques: Expressing, Derivative Assay, Staining, Generated

Journal: Frontiers in Veterinary Science

Article Title: Investigation of activation-induced markers (AIM) in porcine T cells by flow cytometry

doi: 10.3389/fvets.2024.1390486

Figure Lengend Snippet: Expression of AIMs in blood-derived CD4/CD8α-defined T cell subsets after stimulation with SEB. (A) Representative FCM plots depicting expression of CD25, CD69, CD40L and TNF-α in CD4 − CD8α + , CD4 + CD8α + and CD4 + CD8α − T cells. PBMC samples were unstimulated (Medium, left columns) or stimulated with SEB (SEB, right columns) for 18 h. Cells were pre-gated in the following order: live, single and CD3 + . Gates shown indicate gating applied to calculate frequencies of AIM + and TNF-α + cells. (B) Frequencies of CD25 + , CD69 + , CD40L + and TNF-α + cells within CD4/CD8α-defined T cell subsets in unstimulated (Medium) and SEB-stimulated samples. Each dot represents data from one animal ( n = 5). Asterisks indicate significant differences between groups ( * p ≤ 0.05, ** p ≤ 0.01).

Article Snippet: Cells were surface-stained with primary monoclonal antibodies (mAbs) directed against CD3 (PerCP-Cy5.5-conjugated, mouse IgG2a anti-pig, clone: BB23-8E6-8C8, BD Biosciences), CD4 (FITC-conjugated, mouse IgG1 anti-pig, clone: b38c6c7, Bio-Rad Laboratories Ltd., Hercules, CA, United States), CD8α (biotinylated, mouse IgG2a anti-pig, clone: 76–2-11, Southern Biotech, Birmingham, AL, United States), CD25 (AlexaFluor 647-conjugated, mouse IgG1 anti-pig, clone: K231.3B2, Bio-Rad) and CD69 [unconjugated, mouse IgG2b anti-pig, clone: 01-14-22-51 ( )].

Techniques: Expressing, Derivative Assay

Journal: Frontiers in Veterinary Science

Article Title: Investigation of activation-induced markers (AIM) in porcine T cells by flow cytometry

doi: 10.3389/fvets.2024.1390486

Figure Lengend Snippet: Expression of AIMs in blood-derived CD4/CD8α-defined T cell subsets after stimulation with ASFV Estonia 2014. (A) Representative FCM plots depicting expression of CD25, CD69, CD40L, ICOS and TNF-α in CD4 − CD8α + , CD4 + CD8α + and CD4 + CD8α − T cells. PBMC samples were incubated with mock inoculum (Mock, left columns) or ASFV Estonia 2014 (ASFV, right columns) for 18 h. Cells were pre-gated in the following order: live, single and CD3 + . Gates shown indicate gating applied to calculate frequencies of AIM + and TNF-α + cells. Dot size was enlarged to improve visibility of AIM + and TNF-α + cells. (B) Frequencies of CD25 + , CD69 + , CD40L + , ICOS + and TNF-α + cells within CD4/CD8α-defined T cell subsets in mock-inoculated (Mock) and ASFV Estonia 2014 (ASFV)-stimulated samples. Each dot represents data from one animal (n = 5). Asterisks indicate significant differences between groups ( * p ≤ 0.05, ** p ≤ 0.01).

Article Snippet: Cells were surface-stained with primary monoclonal antibodies (mAbs) directed against CD3 (PerCP-Cy5.5-conjugated, mouse IgG2a anti-pig, clone: BB23-8E6-8C8, BD Biosciences), CD4 (FITC-conjugated, mouse IgG1 anti-pig, clone: b38c6c7, Bio-Rad Laboratories Ltd., Hercules, CA, United States), CD8α (biotinylated, mouse IgG2a anti-pig, clone: 76–2-11, Southern Biotech, Birmingham, AL, United States), CD25 (AlexaFluor 647-conjugated, mouse IgG1 anti-pig, clone: K231.3B2, Bio-Rad) and CD69 [unconjugated, mouse IgG2b anti-pig, clone: 01-14-22-51 ( )].

Techniques: Expressing, Derivative Assay, Incubation

Journal: Frontiers in Veterinary Science

Article Title: Investigation of activation-induced markers (AIM) in porcine T cells by flow cytometry

doi: 10.3389/fvets.2024.1390486

Figure Lengend Snippet: Relative distribution of AIM phenotypes within CD4/CD8α-defined T cell subsets in SEB-stimulated vs. ASFV-stimulated PBMC cultures. (A) Doughnut charts of AIM phenotypes in SEB-stimulated samples generated by Boolean gating in CD4 − CD8α + (top row), CD4 + CD8α + (middle row) and CD4 + CD8α − (bottom row) T cells. Each doughnut represents the PBMC sample of one pig. Different phenotypes are indicated by different colors with all AIM phenotypes co-expressing TNF-α summarized in gray. CD25 − CD40L − CD69 − TNF-α − and CD25 single + T cell phenotypes are not shown. (B) Doughnut charts of AIM phenotypes in ASFV Estonia 2014-stimulated samples generated by Boolean gating in CD4 − CD8α + (top row), CD4 + CD8α + (middle row) and CD4 + CD8α − (bottom row) T cells. Each doughnut represents the PBMC sample of one pig. Different phenotypes are indicated by different colors with all AIM phenotypes co-expressing TNF-α summarized in gray. CD25 − CD40L − CD69 − TNF-α − and CD25 single + T cell phenotypes are not shown.

Article Snippet: Cells were surface-stained with primary monoclonal antibodies (mAbs) directed against CD3 (PerCP-Cy5.5-conjugated, mouse IgG2a anti-pig, clone: BB23-8E6-8C8, BD Biosciences), CD4 (FITC-conjugated, mouse IgG1 anti-pig, clone: b38c6c7, Bio-Rad Laboratories Ltd., Hercules, CA, United States), CD8α (biotinylated, mouse IgG2a anti-pig, clone: 76–2-11, Southern Biotech, Birmingham, AL, United States), CD25 (AlexaFluor 647-conjugated, mouse IgG1 anti-pig, clone: K231.3B2, Bio-Rad) and CD69 [unconjugated, mouse IgG2b anti-pig, clone: 01-14-22-51 ( )].

Techniques: Generated, Expressing