Journal: The veterinary quarterly

Article Title: Holstein × Montbéliarde-sired F1 generation crossbred female calves have an increased cellular immune response potential compared with purebred Holsteins.

doi: 10.1080/01652176.2024.2435982

Figure Lengend Snippet: Figure 2. Gating strategies to define bovine T cell subsets and their IFN-γ expression. Mononuclear cells were isolated from blood of 3 months-old female calf and surface stained with antibody cocktails (CD4, CD8, CD44, IFN-γ) after PMA/ionomycin mixture stimulation. The lymphocytes are chosen with FSC versus SSC (A) and then singlets are gated using FSC-a and FSC-H (B). By outputting CD4 versus CD8, CD4+ CD8-, CD8+ CD4- are identified, respectively (C). The CD4+ CD8- T cells expressing IFN-γ (D). The CD8+ CD4- T cells expressing IFN-γ (G). CD4+ CD8- T cells and CD4+ CD8- T cells can be further subdivided into CD44+ and CD44- subsets (E and F), respectively. The lymphocytes expressing IFN-γ (H).

Article Snippet: Monoclonal antibodies (mAb) specific for bovine TCRγδ (WC1), CD8, CD4 and IFN-γ with different fluorochrome conjugates were purchased from BIO-RAD (BIO-RAD, Shanghai, China) and mAb to human CD27, mouse CD44 were purchased from Biolegend (Biolegend, San Diego, USA) (Table 1).

Techniques: Expressing, Isolation, Staining

Journal: The veterinary quarterly

Article Title: Holstein × Montbéliarde-sired F1 generation crossbred female calves have an increased cellular immune response potential compared with purebred Holsteins.

doi: 10.1080/01652176.2024.2435982

Figure Lengend Snippet: Figure 5. Differences of IFN-γ expression in lymphocytes and their T cells of MH and HO calves. Representative dot-plots depict IFN-γ+ T cell subsets in CD4+ (a), CD8+ (B) T cells and total lymphocytes (C). Representative histograms showed the expressions of IFN-γ in lymphocytes (D), CD4+ (E) and CD8+ (F) T cells in the whole blood of MH and HO calves. Data were shown in LSM ± SEM (n = 25). *p < 0.05, **p < 0.01.

Article Snippet: Monoclonal antibodies (mAb) specific for bovine TCRγδ (WC1), CD8, CD4 and IFN-γ with different fluorochrome conjugates were purchased from BIO-RAD (BIO-RAD, Shanghai, China) and mAb to human CD27, mouse CD44 were purchased from Biolegend (Biolegend, San Diego, USA) (Table 1).

Techniques: Expressing

Journal: The veterinary quarterly

Article Title: Holstein × Montbéliarde-sired F1 generation crossbred female calves have an increased cellular immune response potential compared with purebred Holsteins.

doi: 10.1080/01652176.2024.2435982

Figure Lengend Snippet: Figure 6. The expression of IFN-γ was derived from subsets of CD4+ and CD8− T cells, characterized by CD44+ cells rather than CD44− cells. Representative dot-plots depict CD4+ (A) and CD8- (B) T cells subsets of IFN-γ and CD44 from MH (left panel) and HO (right panel) calves. Numbers represent the percentages of cells in each quadrant. Bar graphs showed the expressions of IFN-γ by CD44+ or CD44− subsets in CD4+ (C) and CD8+ (D) T cells of MH and HO calves. Data were shown in LSM ± SEM (n = 25). Labels (a, b, c and d) of different letters indicated a significant difference between any two different sets of data (p < 0.01).

Article Snippet: Monoclonal antibodies (mAb) specific for bovine TCRγδ (WC1), CD8, CD4 and IFN-γ with different fluorochrome conjugates were purchased from BIO-RAD (BIO-RAD, Shanghai, China) and mAb to human CD27, mouse CD44 were purchased from Biolegend (Biolegend, San Diego, USA) (Table 1).

Techniques: Expressing, Derivative Assay

Journal: Journal of Veterinary Science

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine

doi: 10.4142/jvs.2014.15.1.99

Figure Lengend Snippet: Humoral immune responses in vaccinated piglets from the different groups detected with a commercial ELISA kit.

Article Snippet: To further evaluate cellular immune responses, IFN-γ levels in peripheral blood isolated from the vaccinated piglets at 35 dpi were measured using a commercial ELISA kit (Pig Interferon-γ, IFN-γ ELISA Kit; CUSABIO, USA) according to the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Theranostics

Article Title: Nintedanib enhances the efficacy of PD-L1 blockade by upregulating MHC-I and PD-L1 expression in tumor cells

doi: 10.7150/thno.65828

Figure Lengend Snippet: Analysis of mechanisms related to immune activation. (A) Heat map of the expression of signature genes of interferon-gamma (IFN-γ) response (B) quantified by signature score. (C) Heat map of the expression of signature genes of IFN-α response (D) quantified by signature score. Gene Set Enrichment Analysis (GSEA) plot and gene sets for (E) RESPONSE TO INTERFERON-GAMMA and (F) ANTIGEN PROCESSING AND PRESENTATION. Flow cytometry analysis of (G) major histocompatibility complex class I (MHC-I) and (H) PD-L1 expression after different treatments in MC38 cells. The MHC-I expression levels were evaluated by MFI in (I) MC38, (J) LLC, (K) 4T1, and (O) B16F10 cells. The PD-L1 expression levels were evaluated by MFI in (L) MC38, (M) LLC, (N) 4T1, and (P) B16F10 cells. (Q) Representative western blot images of p-STAT3/STAT3, transporter associated with antigen processing 1 (TAP1), PD-L1 and beta-2-microglobulin (β2M).

Article Snippet: Recombinant mouse interferon-gamma (rMuIFN-γ; HY-P7071) and recombinant human interferon-gamma (rHuIFN-γ; HY-P7025) were purchased from MedChem Express (Monmouth Junction, NJ, USA).

Techniques: Activation Assay, Expressing, Flow Cytometry, Western Blot

Journal: Frontiers in Immunology

Article Title: IL-15 Harnesses Pro-inflammatory Function of TEMRA CD8 in Kidney-Transplant Recipients

doi: 10.3389/fimmu.2017.00778

Figure Lengend Snippet: IL-15-stimulated TEMRA CD8 from Kidney-Transplant (KT) Recipients induces the activation of endothelium through IFN-γ and TNF-α secretion. (A) Expression of CD25 or CD69 by CD8 T cell subsets purified from KT recipients stimulated for 24 h with plate-bound anti-CD3 (2 µg/mL) and IL-15 (10 ng/mL). Representative flow data are shown, and bars indicate mean ± SEM of pooled data ( n = 10–12). (B) Schematic overview of the strategy to assess the inflammation of human umbilical vein endothelial cell (HUVEC) by IL-15-activated CD8 T cell subsets from KT. (C) Expression of CX3CL1 by HUVEC after 6 h of culture in the presence of medium control or selective inhibitors [anti-IFN-γ (10 µg/mL) or anti-TNF-γ (10 µg/mL) mAb] and with supernatant (dilution of supernatant of 1/6) from CD8 T cell subsets from KT stimulated overnight with plate-bound anti-CD3 (2 µg/mL) and IL-15 (10 ng/mL). Bars indicate mean ± SEM of pooled data ( n = 4–6). Mann–Whitney test (A) or Kruskal–Wallis test followed by a Dunn’s Multiple Comparison Test (C) was performed (* p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: Antibodies against the following proteins were purchased from BD Biosciences: CD3 (HIT3a and UCHT1), CD69 (SK7), CD25 (M-A251), CD28 (CD28.2), pSTAT5 pY694 (clone 47), p38MAPK pT180/pY182 (38/p38), TNFα (Mab11), IFN-γ (B27), and Annexin V. Antibodies against the following proteins were purchased from Miltenyi Biotech: CD8 (BW135/80), CD45RA (T6D11), CCR7 (REA108), pAKT pS473 (REA359), and pERK1/2 pT202/pY204 (REA152).

Techniques: Activation Assay, Expressing, Purification, Control, MANN-WHITNEY, Comparison