Journal: Scientific Reports

Article Title: Enhancing cancer immunotherapy using cordycepin and C ordyceps militaris extract to sensitize cancer cells and modulate immune responses

doi: 10.1038/s41598-024-72833-x

Figure Lengend Snippet: Changes in cytokines, cytotoxic mediators, and subpopulations of effector immune cells following treatment with cordycepin (CD) and Cm-EE. Non-adherent cells were exposed to 100 µM of cordycepin (CD) or 100 µg/mL Cm-EE for 24 h. Culture supernatants were collected for determination of cytokine and cytotoxic mediator production by using cytokine bead array. The fold changes in cytokine ( a ) and cytotoxic mediator ( b ) production from the cordycepin or Cm-EE treatments were compared to the appropriate control. Subpopulations of treated immune cells ( c ), including CD4 (CD3 + and CD4+), CD8 (CD3 + and CD8+), NK cells (CD3- and CD56+), and B cells (CD3- and CD19+) were characterized using specific fluorescence dye-conjugated monoclonal antibodies. Data are presented as the mean ± SEM. Statistically significant values (* p < 0.05) were determined by Student’s t-test ( N = 3).

Article Snippet: The non-adherent cells were seeded at a density of 1 × 10 6 cells/well in 12-well plate and treated with either 100 µM cordycepin or 100 µg/mL Cm-EE for 24 h. Following treatment, cells were harvested and stained with monoclonal fluorescent-conjugated antibodies, including anti-CD3 FITC (Clone: UCHT-1), anti-CD4 APC (Clone: OKT-4), anti-CD8 APC (Clone: UCHT-4), anti-CD16 APC (Clone: LNK16), anti-CD19 APC (Clone: LT19) (all sourced from Immunotools, Friesoythe, Germany), and anti-CD56 PE (Clone: HCD56; BioLegend).

Techniques: Control, Fluorescence

Journal: AIDS Research and Human Retroviruses

Article Title: T Regulatory Cell Induced Foxp3 Binds the IL2, IFN γ, and TNF α Promoters in Virus-Specific CD8 + T Cells from Feline Immunodeficiency Virus Infected Cats

doi: 10.1089/aid.2017.0187

Figure Lengend Snippet: IL2, IFNγ, and TNFα mRNA from virus-specific CD8+ T cells following autologous CD4+CD25+ Treg cell coculture. Virus-nonspecific and virus-specific CD8+ T cells from FIV+ cats were sorted as described in Figure 1. Virus-nonspecific CD8+ T cells (black bars, CFSEhi) from FIV+ cats were cultured with autologous Treg cells (striped bars, CFSEhi+Treg) and virus-specific CD8+ T cells from FIV+ cats (white bars, CFSElow) were sorted with autologous Treg cells (gray bars, CFSElow+Treg). Cells were isolated and cultured at 1 week (left column), 4 weeks (middle column), and 8 weeks (right column) postinfection. (A) Following Treg cell coculture, IL2 mRNA is decreased in virus-specific CD8+ T cells at 4 and 8 weeks postinfection. (B) Following Treg cell coculture, IFNγ mRNA is decreased in virus-specific CD8+ T cells at 4 and 8 weeks postinfection. (C) TNFα is decreased in virus-specific CD8+ T cells at 4 weeks postinfection following Treg cell coculture (n = 3–7 cats for each time point). Bars represent the mean + SEM. Data were analyzed using an unpaired t-test with significance set at p < .05, asterisks. SEM, standard error of the mean.

Article Snippet: 23 Single cells from LNs were suspended at 1 × 10 8 cells/ml in HBSS with 2% FBS and stained with anti-feline CD8 PE antibody (clone 3.357) at 4°C for 30 min. EasySep ® PE Selection Cocktail was added at 100 μL/ml of cell suspension at room temperature for 15 min, and EasySep Magnetic Nanoparticles were added at 50 μL/ml at RT for 10 min. CD8 + PE + cells were separated by using the magnet provided in the kit (STEMCELL, Vancouver, BC, Canada).

Techniques: Virus, Cell Culture, Isolation

Journal: AIDS Research and Human Retroviruses

Article Title: T Regulatory Cell Induced Foxp3 Binds the IL2, IFN γ, and TNF α Promoters in Virus-Specific CD8 + T Cells from Feline Immunodeficiency Virus Infected Cats

doi: 10.1089/aid.2017.0187

Figure Lengend Snippet: Isolation of virus-specific CD8+ T cells from FIV+ cats. CD8+ T cells were isolated from peripheral LNs of FIV- control cats (A) and FIV+ cats (B). The cells were CFSE stained and returned to CD4+CD25+ depleted LN cultures. The cultures were stimulated with UV inactivated FIV for 72 h and resorted. CD8+ T cells from FIV- control cats exhibited some CFSE dilution in culture (circled, A). Dead cells were gated by DAPI exclusion (not shown). Virus-specific CD8+ T cells from FIV+ cats were identified by CFSE dilution and increased forward scatter (single circle, B) as compared with virus-nonspecific CD8+ T cells that did not divide in response to FIV (double circle, B). CFSE, carboxyfluorescein succinimidyl ester; LN, lymph node; FIV, feline immunodeficiency virus.

Article Snippet: 23 Single cells from LNs were suspended at 1 × 10 8 cells/ml in HBSS with 2% FBS and stained with anti-feline CD8 PE antibody (clone 3.357) at 4°C for 30 min. EasySep ® PE Selection Cocktail was added at 100 μL/ml of cell suspension at room temperature for 15 min, and EasySep Magnetic Nanoparticles were added at 50 μL/ml at RT for 10 min. CD8 + PE + cells were separated by using the magnet provided in the kit (STEMCELL, Vancouver, BC, Canada).

Techniques: Isolation, Virus, Control, Staining

Journal: AIDS Research and Human Retroviruses

Article Title: T Regulatory Cell Induced Foxp3 Binds the IL2, IFN γ, and TNF α Promoters in Virus-Specific CD8 + T Cells from Feline Immunodeficiency Virus Infected Cats

doi: 10.1089/aid.2017.0187

Figure Lengend Snippet: Foxp3 mRNA expression is increased in virus-specific CD8+ T cells following coculture with lentivirus-activated Treg cells. Virus-nonspecific and virus-specific CD8+ T cells from FIV+ cats were sorted as described in Figure 1. Virus-nonspecific CD8+ T cells (black bars, CFSEhi) from FIV+ cats were cultured with autologous Treg cells (striped bars, CFSEhi+Treg) and virus-specific CD8+ T cells from FIV+ cats (white bars, CFSElow) were sorted with autologous Treg cells (gray bars, CFSElow+Treg). Cells were isolated and cultured at 1 week (left), 4 weeks (middle), and 8 weeks (right) postinfection. Following Treg coculture, Foxp3 is increased in virusspecific CD8+ T cells at all time points and in nonspecific CD8+ T cells at week 1 postinfection (n = 4–6 cats for each time point). Bars represent the mean + SEM. Data were analyzed using an unpaired t-test with significance set at p < .05, asterisks.

Article Snippet: 23 Single cells from LNs were suspended at 1 × 10 8 cells/ml in HBSS with 2% FBS and stained with anti-feline CD8 PE antibody (clone 3.357) at 4°C for 30 min. EasySep ® PE Selection Cocktail was added at 100 μL/ml of cell suspension at room temperature for 15 min, and EasySep Magnetic Nanoparticles were added at 50 μL/ml at RT for 10 min. CD8 + PE + cells were separated by using the magnet provided in the kit (STEMCELL, Vancouver, BC, Canada).

Techniques: Expressing, Virus, Cell Culture, Isolation

Journal: AIDS Research and Human Retroviruses

Article Title: T Regulatory Cell Induced Foxp3 Binds the IL2, IFN γ, and TNF α Promoters in Virus-Specific CD8 + T Cells from Feline Immunodeficiency Virus Infected Cats

doi: 10.1089/aid.2017.0187

Figure Lengend Snippet: Foxp3 binds the IL2, IFNγ, and TNFα promoters in virus-specific CD8+ T cells. CD8+ T cells from FIV- cats were cocultured with autologous CD4+CD25+ Treg cells (left column). Virus-specific CD8+ T cells from FIV+ cats 8 weeks (center column) and 6 months (right column) were either untreated (white bars, CFSElow) or cocultured with autologous CD4+CD25+ Treg cells (CFSElow+Treg). After 24 h, Foxp3 ChIP followed by quantitative polymerase chain reaction for IL2, IFNγ, and TNFα promoters were performed. Results show increased Foxp3 binding to the IL2, IFNγ, and TNFα promoters and (A–C) after coculture with autologous T reg cells at both 8 weeks and 6 months postinfection for FIV+ cats (n = 5–7 cats). FIV negative cats exhibited almost no Foxp3 binding to the various promoters following autologous Treg cell coculture. Bars represent the mean + SEM. Data were analyzed using an unpaired t-test with significance set at p < .05, asterisks. ChIP, chromatin immunoprecipitation.

Article Snippet: 23 Single cells from LNs were suspended at 1 × 10 8 cells/ml in HBSS with 2% FBS and stained with anti-feline CD8 PE antibody (clone 3.357) at 4°C for 30 min. EasySep ® PE Selection Cocktail was added at 100 μL/ml of cell suspension at room temperature for 15 min, and EasySep Magnetic Nanoparticles were added at 50 μL/ml at RT for 10 min. CD8 + PE + cells were separated by using the magnet provided in the kit (STEMCELL, Vancouver, BC, Canada).

Techniques: Virus, Real-time Polymerase Chain Reaction, Binding Assay, Chromatin Immunoprecipitation

Journal: The Journal of Infectious Diseases

Article Title: Serology of Severe Acute Respiratory Syndrome: Implications for Surveillance and Outcome

doi: 10.1086/380397

Figure Lengend Snippet: Relationship between CD3 + CD4 + and CD3 + CD8 + T cell percentages and presence of IgG and IgM antibodies, in patients with probable severe acute respiratory syndrome coronavirus (SARS CoV) infection.

Article Snippet: In brief, samples of whole blood were collected in EDTA and were immunolabeled with monoclonal antibodies CD4-FITC, CD8- PE, and CD3-CY5 (Immunotech) at room temperature; mouse IgG1-FITC, mouse IgG1-PE, and mouse IgG1-CY5 (Immunotech) were incubated as isotype controls, to allow for subtraction of nonspecific staining.

Techniques: Infection

Journal: PLoS ONE

Article Title: Effect of age on chronic inflammation and responsiveness to bacterial and viral challenges

doi: 10.1371/journal.pone.0188881

Figure Lengend Snippet: (A) Differences in the proportions of monocytes, T cells and NK cells with age. (B) Differences with age in the % of T cells that are CD4+ and CD8+, in the CD4/CD8 ratio with age, in the proportion of T cells that are Tregs, and in the proportion of CD4 and CD8 cells that are CD28+ and -. (C) Anti-CD3 + anti-CD28 stimulated proliferation (left panel) and IFNγ production (right panel). Results are expressed as the mean ± SEM. * indicates a statistically significant difference (P <0.05) between the young and old cohorts.

Article Snippet: The antibodies used were: CD14-PE (clone MØp9), CD8-PE (clone SK1) and CD3-FITC (clone SK7) from StemCell Technologies, Vancouver, BC; CD45-FITC (Hle1), CD28-APC (clone CD28.2), CD4-PE-Cy7 (clone SK3), CD25-BB515 (clone 2A3), CD127-AF647 (clone HIL-7R-M21) and FoxP3-PE (clone 236A/E7) from BD Biosciences, Mississauga, ON; CD56-APC (clone CMSSB) from eBioscience, San Diego, CA.

Techniques:

Journal: Cell reports

Article Title: Single-Cell Mapping of Progressive Fetal-to-Adult Transition in Human Naive T Cells

doi: 10.1016/j.celrep.2020.108573

Figure Lengend Snippet: Fetal splenic, full-term UCB, and adult PB naive CD4 (n = 15) and CD8 (n = 13) T cell samples were profiled by single-cell RNA sequencing (scRNA-seq). See also and . (A) UMAP plot showing distribution of cells of each age and CD4-versus-CD8 lineages. Separate plots along the bottom and right show cells of individual identities. (B) UMAP plot showing Louvain clustering of single-cell profiles. (C and D) Quantification of CD4 (C) and CD8 (D) T cell clustering where each column represents a different sample. A, adult; F, fetal; U, UCB. (E) Overview of single-cell developmental stage score model generation. (F and G) Developmental stage scoring of individual CD4 (F) and CD8 (G) naive T cells for each sample. Points are individual cells.

Article Snippet: mouse anti-human CD8-PE monoclonal antibody (clone SK1) , BD Biosciences , CT#345773; RRID: AB_2868801.

Techniques: RNA Sequencing Assay

Journal: Cell reports

Article Title: Single-Cell Mapping of Progressive Fetal-to-Adult Transition in Human Naive T Cells

doi: 10.1016/j.celrep.2020.108573

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: mouse anti-human CD8-PE monoclonal antibody (clone SK1) , BD Biosciences , CT#345773; RRID: AB_2868801.

Techniques: Recombinant, Isolation, Selection, Cell Isolation, Amplification, Binding Assay, Microarray, Software, Real-time Polymerase Chain Reaction