Journal: Brain, Behavior, & Immunity - Health

Article Title: Stress hormones are associated with inflammatory cytokines and attenuation of T-cell function in the ascites from patients with high grade serous ovarian cancer

doi: 10.1016/j.bbih.2022.100558

Figure Lengend Snippet: Ascites-derived T-cells exhibit a heterogeneous activation and inhibitory receptors pattern, while epinephrine reduces the expression of markers associated with T-cell function. (A) Protein-protein interaction network between cytokines positively correlated with stress hormones/metabolites. (B) Bioinformatic analyses utilizing the GO biological processes based on the protein-protein interaction of stress hormone correlated cytokines. (C) Cellular expression of activation and immune checkpoint markers on T-cells isolated from ovarian ascites. 14-color polychromatic flow cytometry analysis was performed on ascites mononuclear cells. Gated CD4 and CD8 T-cells are shown. (D) Ascites derived T-cells from patients with ovarian cancer (n = 4) stimulated with αCD3 and αCD28 for seven days in the presence and absence of epinephrine. Flow cytometry analysis of Granzyme B and CD38 co-expression. Quantification from individual patients showing the frequency of Granzyme B and CD38 co-expression ( P = 0.0042). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Primary antibodies for CD4 (552051, BD Biosciences), CD8 (MCD0801, Bio-Analytical), and PD-1 (75781-958, VWR) were added for an hour.

Techniques: Derivative Assay, Activation Assay, Expressing, Cell Function Assay, Isolation, Flow Cytometry

Journal: Brain, Behavior, & Immunity - Health

Article Title: Stress hormones are associated with inflammatory cytokines and attenuation of T-cell function in the ascites from patients with high grade serous ovarian cancer

doi: 10.1016/j.bbih.2022.100558

Figure Lengend Snippet: Daily restraint stress leads to increased CD4 + PD-1 + /CD8 + PD-1 + T-cell ratio in the ovarian tumor microenvironment. (A) Density plot for lymphocyte size selection. (B) Histograms of CD4 + , CD8 + and PD-1 + staining distribution. (C) . Cellular expression of CD4 + PD-1 + and CD8 + PD-1 + cells measured by flow cytometry analysis on ID8 tumor samples from mice subjected to daily and controls. (D) Quantification of CD4 + PD-1 + and CD8 + PD-1 + cellular expression. (E) Quantification of CD4 + PD-1 + /CD8 + PD-1 + T-cell ratio indicates a significant increase ( p = 0.0360) in the daily restraint stress group.

Article Snippet: Primary antibodies for CD4 (552051, BD Biosciences), CD8 (MCD0801, Bio-Analytical), and PD-1 (75781-958, VWR) were added for an hour.

Techniques: Selection, Staining, Expressing, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: The Inhibitory T Cell Receptors PD1 and 2B4 Are Differentially Regulated on CD4 and CD8 T Cells in a Mouse Model of Non-alcoholic Steatohepatitis

doi: 10.3389/fphar.2019.00244

Figure Lengend Snippet: T cell characterization by flow cytometry in the blood and livers of chow and WD fed mice. (A) T cells (CD45+Hoechst-CD3+NK1.1-) in the blood and liver after SC feeding. ( n = 5, ∗∗ p < 0.01) (B) Basic PD1 and 2B4 expression in blood and liver CD45+CD4+ and CD45+CD8+ T cells under SC. ( n = 5, ∗∗ p < 0.01) (C) Absolute leukocyte cell numbers in blood and liver after SC and WD feeding. ( n = 5). (D) Absolute T cell numbers in blood and liver after feeding a SC or WD for 24 weeks. ( n = 5, ∗ p < 0.05) (E) Absolute CD4+ and CD8+ T cell numbers after SC feeding and after a metabolic challenge by a 24 week fed WD. ( n = 5, ∗ p < 0.05 and ∗∗ p < 0.01) (F) Percentage of CD107a positive CD8+ T cells in blood and liver tissue after administration of a SC or WD for 24 weeks. ( n = 5, ∗∗∗ p < 0.001).

Article Snippet: Afterward the sections were incubated with the primary antibody against CD4 (Biozol, cat. no.: bs-0647R) for 1 h in a humidified chamber at room temperature, followed by detection using the anti-rabbit (Cell Signaling Technology, cat. no.: 7074S).

Techniques: Flow Cytometry, Expressing

Journal: Frontiers in Pharmacology

Article Title: The Inhibitory T Cell Receptors PD1 and 2B4 Are Differentially Regulated on CD4 and CD8 T Cells in a Mouse Model of Non-alcoholic Steatohepatitis

doi: 10.3389/fphar.2019.00244

Figure Lengend Snippet: Expression of inhibitory T cell receptors PD1 and 2B4 increases on hepatic T cells under WD. (A) Blood and intrahepatic PD1 expressing CD8+ cells were analyzed by Flow Cytometry after SC and WD feeding for 24 weeks. ( n = 5, ∗ p < 0.05 and ∗∗∗ p < 0.001). (B) Percentage of intrahepatic degranulating CD107a+ CD8 T cells of PD1+ and PD1- cells after SC or WD treatment for 24 weeks. ( n = 5, ∗ p < 0.05). (C) Upregulation of PD1 on intrahepatic CD8+ CD62L+ (Naïve + central memory CD8+ T cells) and CD8+ CD62L- (effector memory CD8+ T cells) T cells after SC or WD. ( n = 5, ∗ p < 0.05). (D) 2B4 expression on CD8+ T cells in blood and liver after SC or WD feeding for 24 weeks. ( n = 5, ∗ p < 0.05). (E) PD1 expression on blood and hepatic CD4+ T cells under SC or WD for 24 weeks. ( n = 5). (F) Inhibitory receptor 2B4 expression blood and intrahepatic CD4 T cells under SC or WD. ( n = 5, ∗ p < 0.05).

Article Snippet: Afterward the sections were incubated with the primary antibody against CD4 (Biozol, cat. no.: bs-0647R) for 1 h in a humidified chamber at room temperature, followed by detection using the anti-rabbit (Cell Signaling Technology, cat. no.: 7074S).

Techniques: Expressing, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: The Inhibitory T Cell Receptors PD1 and 2B4 Are Differentially Regulated on CD4 and CD8 T Cells in a Mouse Model of Non-alcoholic Steatohepatitis

doi: 10.3389/fphar.2019.00244

Figure Lengend Snippet: Activation markers CD69 and CD44 are increased under metabolic stress on hepatic CD4 T cells, but not CD8 T cells, while this activation is inhibited in PD1+ CD4 T cells. (A) CD25 expression on blood and liver CD8 T cells under SC or WD for 24 weeks. ( n = 5). (B) CD69 expression on blood and on hepatic CD8 T cells after 24 weeks treatment with SC or WD. ( n = 5), ( ∗∗∗ p < 0.001). (C) CD69 expression on blood and hepatic CD4 T cells after 24 weeks SC and WD feeding. ( n = 5, ∗ p < 0.05). (D) CD44 expression on blood and intrahepatic CD4 T cells under SC or WD treatment for 24 weeks. ( n = 5, ∗∗ p < 0.01 and ∗∗∗ p < 0.001). (E) CD25 expression on blood and hepatic CD4 T cells under SC or WD. ( n = 5), ( ∗ p < 0.05 and ∗∗∗ p < 0.001). (F) CD44 and CD69 expression on PD1+ or PD1- CD4 T cells after SC or WD feeding. ( n = 5), ( ∗ p < 0.05; ∗∗ p < 0.01; and ∗∗∗ p < 0.001).

Article Snippet: Afterward the sections were incubated with the primary antibody against CD4 (Biozol, cat. no.: bs-0647R) for 1 h in a humidified chamber at room temperature, followed by detection using the anti-rabbit (Cell Signaling Technology, cat. no.: 7074S).

Techniques: Activation Assay, Expressing

Journal: Frontiers in Pharmacology

Article Title: The Inhibitory T Cell Receptors PD1 and 2B4 Are Differentially Regulated on CD4 and CD8 T Cells in a Mouse Model of Non-alcoholic Steatohepatitis

doi: 10.3389/fphar.2019.00244

Figure Lengend Snippet: PD1+ and 2B4+ infiltrating cells are increased in NASH patients. (A) Intrahepatic PD1+ CD4 or CD8+ T cells and (B) 2B4+ CD4 or CD8+ T cells were analyzed by flow cytometry. Analysis include 5 control and 5 NASH patients. Intrahepatic cells were gated FSC/SSC, duplets were excluded, living cells, CD3+, PD1+/CD4+, PD1+/CD8+, 2B4+/CD4+, 2B4+/CD8+. Displayed is the percentage of recorded cells in liver biopsy samples. (C) Intrahepatic PD1+/CD14+/CD68+ and (D) 2B4+/CD14+/CD68+ monocytes were analyzed by flow cytometry. Analysis include 5 control and 5 NASH patients. Intrahepatic cells were gated FSC/SSC, duplets were excluded, living cells, CD14+/CD68+, PD1+ or 2B4+. Displayed is the percentage of recorded cells in liver biopsy samples. ( ∗ p < 0.05; ∗∗ p < 0.01; and ∗∗∗ p < 0.001).

Article Snippet: Afterward the sections were incubated with the primary antibody against CD4 (Biozol, cat. no.: bs-0647R) for 1 h in a humidified chamber at room temperature, followed by detection using the anti-rabbit (Cell Signaling Technology, cat. no.: 7074S).

Techniques: Flow Cytometry, Control