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