Journal: Cell reports

Article Title: Consumption of fish oil high-fat diet induces murine hair loss via epidermal fatty acid binding protein in skin macrophages

doi: 10.1016/j.celrep.2022.111804

Figure Lengend Snippet: (A) Representative hematoxylin and eosin staining of skin tissues from mice fed the LFD, fish oil HFD, or cocoa butter diet at different magnifications (red arrows: infiltrating inflammatory cells). Scale bars, 100 μm. (B and C) Flow cytometric analysis of immune cell phenotype in the dermis of mice on different diets for 3 months. A cell population with strong autofluorescence (red trapezoid gate) was specifically accumulated in the dermis of mice fed the fish oil HFD (B). Multichannel signals of the autofluorescent cells were analyzed using a BD Fortessa flow cytometer (C). (D–F) Autofluorescent cells in the dermis of fish oil HFD-fed mice were purified using a BD FACSAria II flow sorter and stained with a panel of metal-tagged CyTOF antibodies. Uniform manifold approximation and projection (UMAP) was used to visualize and identify immune cell populations in unsorted dermal cells (D) and sorted autofluorescent dermal cells (E). Individual surface marker signatures in the CyTOF panel are shown in (F). (G and H) Representative IHC images of F4/80 + macrophages (brown staining, G) and CD8 + T cells (brown staining, H) in the skin of mice fed the LFD, fish oil HFD, or cocoa butter HFD. Scale bars, 100 μm. See also . These in vitro experiments were repeated with at least three biological replicates.

Article Snippet: Rat monoclonal anti-mouse CD8a - 168Er , Fluidigm , Cat#3168003B; RRID:AB_2811241.

Techniques: Staining, Flow Cytometry, Purification, Marker, In Vitro

Journal: Cell reports

Article Title: Consumption of fish oil high-fat diet induces murine hair loss via epidermal fatty acid binding protein in skin macrophages

doi: 10.1016/j.celrep.2022.111804

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rat monoclonal anti-mouse CD8a - 168Er , Fluidigm , Cat#3168003B; RRID:AB_2811241.

Techniques: Purification, Recombinant, Activation Assay, SYBR Green Assay, Reverse Transcription, Detection Assay, Enzyme-linked Immunosorbent Assay, Selection, Software

Journal: Translational Oncology

Article Title: RNA demethylase ALKBH5 suppresses tumorigenesis via inhibiting proliferation and invasion and promoting CD8 + T cell infiltration in colorectal cancer

doi: 10.1016/j.tranon.2023.101683

Figure Lengend Snippet: ALKBH5 expression promotes CD8 + T cells infiltration in human CRC tissues. a Infiltration level of 22 immune cells in the ALKBH5 high and low expression groups were examined using CIBERSORT. b Scatterplots show the correlation among activated memory CD4 + T cells, CD8 + T cells, Tregs and the ALKBH5 expression level. c-d Correlation and quantitative analyses of ALKBH5 expression with CD4, CD8 and Foxp3 in human CRC tissues using immunohistochemistry, the data represented mean ± SD (n=56).

Article Snippet: The antibodies used in this study included CD8 (1:1000, ab209775) (Abcam, Cambridge, UK), CD4 (1:1000, ab133636) (Cell Signaling Technology, MA, USA), Foxp3 (1:1000, ab20034) (Abcam, Cambridge, UK).

Techniques: Expressing, Immunohistochemistry

Journal: Translational Oncology

Article Title: RNA demethylase ALKBH5 suppresses tumorigenesis via inhibiting proliferation and invasion and promoting CD8 + T cell infiltration in colorectal cancer

doi: 10.1016/j.tranon.2023.101683

Figure Lengend Snippet: Analysis of the correlations between chemokines and CD8A expression level. a–h CCL5, XCL2, CXCL9, CCL4, CCL3, CXCL10, CCL21 and CXCL12, respectively.

Article Snippet: The antibodies used in this study included CD8 (1:1000, ab209775) (Abcam, Cambridge, UK), CD4 (1:1000, ab133636) (Cell Signaling Technology, MA, USA), Foxp3 (1:1000, ab20034) (Abcam, Cambridge, UK).

Techniques: Expressing

Journal: Translational Oncology

Article Title: RNA demethylase ALKBH5 suppresses tumorigenesis via inhibiting proliferation and invasion and promoting CD8 + T cell infiltration in colorectal cancer

doi: 10.1016/j.tranon.2023.101683

Figure Lengend Snippet: ALKBH5 disrupting NF-κB-CCL5 signaling. a-b Relative expression of CCL5 in ALKBH 5-overexpressed HCT116 and SW620 cells. c Immunohistochemistry was used to detect the expression of ALKBH5, p65, CCL5, and CD8 in human colorectal cancer tissues, the data represented mean ± SD (n=56). d CCL5 expression and the total and phosphorylation level of p65 and IκB-α after ALKBH5 overexpression were examined by western blot analysis. e SW620 were pretreated with 100 and 200 ng/ml of specific NF-κB activator PMA, and western blot analysis was to assess the expression of CCL5 and p-p65.

Article Snippet: The antibodies used in this study included CD8 (1:1000, ab209775) (Abcam, Cambridge, UK), CD4 (1:1000, ab133636) (Cell Signaling Technology, MA, USA), Foxp3 (1:1000, ab20034) (Abcam, Cambridge, UK).

Techniques: Expressing, Immunohistochemistry, Over Expression, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Not all SCID pigs are created equally: Two independent mutations in the Artemis gene cause Severe Combined Immunodeficiency (SCID) in pigs

doi: 10.4049/jimmunol.1501132

Figure Lengend Snippet: The number in each diagram is the percentage of each cell type in thymocyte populations of a SCID pig or a non-SCID carrier littermate (panels A and B). CD8a (x-axis) and CD4 (y-axis; panel A) and γδ (panel B) are T-cell surface markers. The least square means of percentages of each thymocyte sub-population are shown in panel C. Error bars represent the standard error of the estimates. Bars with different letters (a and b) indicate statistically significant (p<0.01) differences between SCID and non-SCID expression within cellular subset.

Article Snippet: Cells were washed in PBS, counted, and 1,000,000 cells were stained using the following fluorescently conjugated antibodies: Alexa Fluor647 conjugated anti-pig CD8a (clone 76-2-11; BD Biosciences, San Diego, California), FITC conjugated anti-pig CD4 (clone 74-12-4; BD Biosciences), PE-conjugated anti-pig γδ (clone MAC320; BD Biosciences), or the appropriate isotype controls (BD Biosciences), or a with FITC-conjugated mouse anti-pig CD3ε (clone PPT3; Southern Biotech, Birmingham, Alabama).

Techniques: Expressing

Journal:

Article Title: Maternal Microchimerism Leads to the Presence of Interleukin-2 in Interleukin-2 Knock Out Mice: Implications for the Role of Interleukin-2 in Thymic Function

doi: 10.1016/j.cellimm.2007.04.002

Figure Lengend Snippet: Serial assessment of thymocyte populations in RAG-1−/− mice reconstituted with either DO11.10/IL-2 WT or DO11.10/IL-2 KO bone marrow. Thymocytes, prepared from mice 6 - 17 weeks post-transplantation, were analyzed by flow cytometry for the presence of (A) CD4 and CD8, or (B) KJ1-26 and CD4. The numbers above the dot plots in (A) represent the number of cells in the thymus from which the dot plot originated. Data shown are representative of 3 or more experiments, with 3 animals per group. Data from the 17-week group are representative of 1 experiment, with 6 animals per group.

Article Snippet: Anti-CD4 (RM4-5), CD8 (Ly-2), CD16/32 (2.4G2), and CD25 (PC61) were purchased from BD Pharmingen (San Diego, CA).

Techniques: Transplantation Assay, Flow Cytometry

Journal:

Article Title: Maternal Microchimerism Leads to the Presence of Interleukin-2 in Interleukin-2 Knock Out Mice: Implications for the Role of Interleukin-2 in Thymic Function

doi: 10.1016/j.cellimm.2007.04.002

Figure Lengend Snippet: Change in frequency of CD4+CD8+ thymocytes, with age, in RAG-KO/DO11.10/IL-2 KO mice. Thymocytes, prepared from 5 or 20 week-old RAG-KO/DO11.10/IL-2 WT or RAG-KO/DO11.10/IL-2 KO mice, were analyzed by flow cytometry for the presence of CD4 and CD8. Data shown are representative of 3 pairs of mice of similar ages.

Article Snippet: Anti-CD4 (RM4-5), CD8 (Ly-2), CD16/32 (2.4G2), and CD25 (PC61) were purchased from BD Pharmingen (San Diego, CA).

Techniques: Flow Cytometry

Journal:

Article Title: Maternal Microchimerism Leads to the Presence of Interleukin-2 in Interleukin-2 Knock Out Mice: Implications for the Role of Interleukin-2 in Thymic Function

doi: 10.1016/j.cellimm.2007.04.002

Figure Lengend Snippet: Phenotype of lymphocyte populations in the spleen and lymph nodes of RAG-1−/− mice reconstituted with either DO11.10/IL-2 WT or DO11.10/IL-2 KO bone marrow. Lymphocytes, prepared from the above organs of mice 12 weeks post-transplantation, were analyzed by flow cytometry for the frequency of CD4+, CD8+ or CD4+CD8+ cells within the KJ1-26+ population. Data shown are representative of 3 or more experiments, with 3 animals per group.

Article Snippet: Anti-CD4 (RM4-5), CD8 (Ly-2), CD16/32 (2.4G2), and CD25 (PC61) were purchased from BD Pharmingen (San Diego, CA).

Techniques: Transplantation Assay, Flow Cytometry

Journal:

Article Title: Maternal Microchimerism Leads to the Presence of Interleukin-2 in Interleukin-2 Knock Out Mice: Implications for the Role of Interleukin-2 in Thymic Function

doi: 10.1016/j.cellimm.2007.04.002

Figure Lengend Snippet: Generation of CD8+KJ1-26+ T cells in thymuses of RAG-1−/− mice reconstituted with either DO11.10/IL-2 WT or DO11.10/IL-2 KO bone marrow. Thymocytes, prepared from chimeras 6, 10, and 17 weeks post-transplantation, were assessed by flow cytometry for the presence of CD8+KJ1-26+ T cells. The dot plots shown represent the distribution of CD4+, CD8+, and CD4+CD8+ thymocytes within the KJ1-26+ thymocyte population. Data shown are representative of 3 or more experiments, with 3 animals per group.

Article Snippet: Anti-CD4 (RM4-5), CD8 (Ly-2), CD16/32 (2.4G2), and CD25 (PC61) were purchased from BD Pharmingen (San Diego, CA).

Techniques: Transplantation Assay, Flow Cytometry

Journal: PLoS ONE

Article Title: Deficient EBV-Specific B- and T-Cell Response in Patients with Chronic Fatigue Syndrome

doi: 10.1371/journal.pone.0085387

Figure Lengend Snippet: (A) Comparison of cytokine producing CD4 + (upper panels) and CD8 + T cells (lower panels) of CFS patients and healthy controls after 10 days of stimulation with EBNA-1 (left panel, Control n = 17, CFS n = 23). Boolean gating strategy was applied to analyze IFN-γ/TNF-α/IL-2 triple, IFN-γ/TNF-α double, and IFN-γ and TNF-α single cytokine producing T cells after intracellular staining of isolated PBMCs incubated with Brefeldin A for 16 h. Stimulation with CMV pp65 (right panel, Control n = 7, CFS n = 5) is shown for IFN-γ/TNF-α/IL-2 triple, and IFN-γ single cytokine producing T cells. (B) Frequencies of PD-1 expression were analyzed for IFN-γ/TNF-α double producing CD4 + and CD8 + T cells after 10 days of stimulation with EBNA-1 or pp65 (n = 8). Statistical analysis was performed using the two-tailed Mann-Whitney-U test with ** p<0.01.

Article Snippet: Live/dead cells were discriminated using an amine reactive dye (Invitrogen, Life Technologies, Darmstadt, Germany) and stained with fluorescence conjugated monoclonal antibodies against CD3, CD4, CD8, PD-1, IFN-y, TNF-α and IL-2 (BD Biosciences, NJ, USA).

Techniques: Staining, Isolation, Incubation, Expressing, Two Tailed Test, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Analysis of the In Vivo Turnover of CD4+ T-Cell Subsets in Chronically SIV-Infected Sooty Mangabeys

doi: 10.1371/journal.pone.0156352

Figure Lengend Snippet: The mean (±SEM) percent Ki67+ (A+B) and percent BrdU+ (C+D) of peripheral blood CD4+ (A+C) and CD8+ (B+D) T-cells were assessed longitudinally by flow cytometry in both uninfected (black squares) and infected (red circles) animals. p = NS between infected and uninfected controls at all timepoints for all populations (Mann-Whitney U). Shaded area represents BrdU administration period. Data points are shown only for animals above which 100 events were collected for the parent population.

Article Snippet: Mouse-derived monoclonal antibodies directed against the following antigens were used at titration-assessed volumes as follows: BrdU (clone 3D4, 20uL), CCR5 (3A9, 10uL), CD3 (SP34-2, 5uL), CD8 (SK1, 5uL), CD62L (SK11, 10uL), Ki67 (B56, 20uL) from BD Biosciences Pharmingen, CD4 (OKT4, 5uL) from BioLegend, CD28 (CD28.2, 10uL) from Beckman-Coulter, and CD95 (DX2, 10uL) from eBioscience.

Techniques: Flow Cytometry, Infection, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Analysis of the In Vivo Turnover of CD4+ T-Cell Subsets in Chronically SIV-Infected Sooty Mangabeys

doi: 10.1371/journal.pone.0156352

Figure Lengend Snippet: A-B. (±SEM) percent BrdU expression was assessed longitudinally by flow cytometry among CD4+ (A) and CD8+ (B) T N , T CM , T TM , T EM , and CCR5+ TM in uninfected (left) and SIV-infected (right) animals. C. Mean (±SEM) longitudinal Ki67 expression of CD4+ T N , T CM , T TM , T EM , and CCR5+ TM in uninfected (left) and SIV-infected (right) animals. p = NS between infected and uninfected controls at all timepoints for all populations (Mann-Whitney U). Shaded area represents BrdU administration period. Data points are shown only for animals above which 100 events were collected for the parent population.

Article Snippet: Mouse-derived monoclonal antibodies directed against the following antigens were used at titration-assessed volumes as follows: BrdU (clone 3D4, 20uL), CCR5 (3A9, 10uL), CD3 (SP34-2, 5uL), CD8 (SK1, 5uL), CD62L (SK11, 10uL), Ki67 (B56, 20uL) from BD Biosciences Pharmingen, CD4 (OKT4, 5uL) from BioLegend, CD28 (CD28.2, 10uL) from Beckman-Coulter, and CD95 (DX2, 10uL) from eBioscience.

Techniques: Expressing, Flow Cytometry, Infection, MANN-WHITNEY