Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Persistent Herpes Simplex Virus Type 1 Infection of Enteric Neurons Triggers CD8 + T Cell Response and Gastrointestinal Neuromuscular Dysfunction

doi: 10.3389/fcimb.2021.615350

Figure Lengend Snippet: Antibodies used in the study.

Article Snippet: CD4 (rabbit) Cy7 conjugate , 50134-R001 , Sino Biological Inc , FC.

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Persistent Herpes Simplex Virus Type 1 Infection of Enteric Neurons Triggers CD8 + T Cell Response and Gastrointestinal Neuromuscular Dysfunction

doi: 10.3389/fcimb.2021.615350

Figure Lengend Snippet: CD3 + cells infiltrate the LMMP following HSV-1 infection. (A) LMMP preparations were obtained from the ileum of sham- and HSV-1-infected mice as described in Methods. The samples were enzymatically digested, and the resulting cell suspensions were labeled with anti-CD11c and anti-F4/80 antibodies and analyzed using flow cytometry. Cells were first selected on a forward scatter and side scatter dot plot and then double positive cells were recorded. The data are reported as the number of CD11c + F4/80 + cells detected in 10 5 events. The experiments were repeated 2 times; n=4 mice per group. (B) Cell suspension obtained as described in (A) were labeled with anti-CD3 antibody and analyzed by flow cytometry. Data are reported as the percentage of CD3 + cells in 10 5 events on side scatter dot plot. The experiments were repeated 2 times; n=4 mice per group. * denotes p < 0.05 vs sham infected mice. (C) Sections of ileum obtained from the sham or HSV-1 infected mice were subjected to immunohistochemistry using anti-CD3 antibody. Scale bars: 50 μm. Representative images of four separate experiments; n=3 mice per group. (D) Cell suspensions obtained from the LMMP, as described in (A) , were labeled with anti-CD3 and anti-CD4 antibodies (E) or with anti-CD3 and anti-CD8 antibodies (F) or with anti-CD3 and anti-CD69 antibodies. The samples were analyzed using flow cytometry. Cells were first selected on a forward scatter and side scatter dot plot and then double positive cells were recorded in 10,000 events. Data are reported as the percentage of double positive cells. Experiments were repeated 3 times n=4 mice per group. * denotes p < 0.05 vs sham infected mice.

Article Snippet: CD4 (rabbit) Cy7 conjugate , 50134-R001 , Sino Biological Inc , FC.

Techniques: Infection, Labeling, Flow Cytometry, Immunohistochemistry

Journal: American Journal of Physiology - Renal Physiology

Article Title: IL-17 mediates neutrophil infiltration and renal fibrosis following recovery from ischemia reperfusion: compensatory role of natural killer cells in athymic rats

doi: 10.1152/ajprenal.00462.2016

Figure Lengend Snippet: Phenotypic analysis of infiltrating mononuclear cells in immune-deficient athymic rats and immune-competent euthymic rats following I/R and exposure to high-salt diet. Lymphocytes were obtained from postischemic rats fed high-salt diet (4.0% NaCl) with or without MMF as labeled. A: gating strategy for FACS analysis. Lymphocytes were gated based on forward and side scatter plot. These were further gated based on the surface expression of CD4, CD8, and CD161. With the use of a histogram, CD4+ cells were further analyzed for expression of IL-17. B and C: no. of CD4+ (B) and CD8+ (C) cells isolated from kidneys. D: no. of activated T cells (CD4+CD25+) isolated from post-I/R or sham-operated rats fed high-salt diet with or without MMF. E–G: no. of CD4+IL-17+ (E), CD8+ natural killer T (NKT) cells (CD8+CD161+) (F), and CD4+NKT (CD4+CD161+) (G) cells isolated from athymic and euthymic rats treated with or without MMF. Data are means ± SE. P < 0.05, injury vs. sham group (*), MMF vs. vehicle ($), and athymic vs. euthymic (#) using ANOVA and Student-Neuman-Keuls post hoc test (n = 5–8 animals/group).

Article Snippet: To evaluate T lymphocytes, the cells were stained with antibodies against rat CD4 (PE-Cy7; BD Biolgend, San Diego, CA), CD8a (Alexa 647; BD Biolgend), CD25 (FITC; BD Biolgend), CD3 (PE; BD Biolegend), and CD161(PerCp; eBiosciences, San Diego, CA).

Techniques: Labeling, Expressing, Isolation

Journal: American Journal of Physiology - Renal Physiology

Article Title: IL-17 mediates neutrophil infiltration and renal fibrosis following recovery from ischemia reperfusion: compensatory role of natural killer cells in athymic rats

doi: 10.1152/ajprenal.00462.2016

Figure Lengend Snippet: Compensatory role of NKT cells in IL-17 production in T cell-deficient postischemic athymic rats fed high-salt diet. Lymphocytes were isolated from postischemic euthymic and athymic rats 2 days post-I/R. The FACS gating strategy is shown in A. In brief, lymphocytes are gated based of their forward and side scatter. These cells are then further subdivided into CD4+ and CD8+ fractions. CD4+ cells were then separated into CD161+ and CD161− cells using a histogram plot. B: schematic diagram for different IL-17-secreting cells by flow cytometry. C: no. of total IL-17+ cells isolated from sham-operated and postischemic kidneys. D and E: total no. of CD4+IL-17+ cells (D) and CD8+IL-17+ cells (E) isolated from postischemic and sham-operated rats. F and G: portion of CD4+IL-17+ cells that are either CD161− (F) or CD161+ (G). H: no. of total IL-17-secreting NKT cells (CD161+) that comprise both CD4+NKT and CD8+NKT. Data are means ± SE. P < 0.05, injury vs. sham group (*) and athymic vs. euthymic ($) using ANOVA and Student-Neuman-Keuls post hoc test (n = 5–6 animals/group).

Article Snippet: To evaluate T lymphocytes, the cells were stained with antibodies against rat CD4 (PE-Cy7; BD Biolgend, San Diego, CA), CD8a (Alexa 647; BD Biolgend), CD25 (FITC; BD Biolgend), CD3 (PE; BD Biolegend), and CD161(PerCp; eBiosciences, San Diego, CA).

Techniques: Isolation, Flow Cytometry

Journal: American Journal of Physiology - Renal Physiology

Article Title: IL-17 mediates neutrophil infiltration and renal fibrosis following recovery from ischemia reperfusion: compensatory role of natural killer cells in athymic rats

doi: 10.1152/ajprenal.00462.2016

Figure Lengend Snippet: Number of IL-17 + cells from different sources isolated postischemic rats

Article Snippet: To evaluate T lymphocytes, the cells were stained with antibodies against rat CD4 (PE-Cy7; BD Biolgend, San Diego, CA), CD8a (Alexa 647; BD Biolgend), CD25 (FITC; BD Biolgend), CD3 (PE; BD Biolegend), and CD161(PerCp; eBiosciences, San Diego, CA).

Techniques: Isolation

Journal: American Journal of Physiology - Renal Physiology

Article Title: IL-17 mediates neutrophil infiltration and renal fibrosis following recovery from ischemia reperfusion: compensatory role of natural killer cells in athymic rats

doi: 10.1152/ajprenal.00462.2016

Figure Lengend Snippet: Effect of systemic IL-17 blockade on infiltrating immune cells in postischemic rats fed high-salt diet. A–D: no. of total CD4 (A), CD8 (B), B cells (Rt1B+) (C), and DC/Mac (CD11b/c+) (D) isolated from post-AKI kidney of rats treated with IL-17Rc antagonist or vehicle. E: representative images of neutrophil staining in kidneys from vehicle or IL-17Rc-treated rats. Magnification is shown. F: quantification of neutrophil infiltration (%positive area) in renal medulla. Data are means ±SE. *P < 0.05, IL-17Rc vs. vehicle using Student’s t-test (n = 3-5 animals/group).

Article Snippet: To evaluate T lymphocytes, the cells were stained with antibodies against rat CD4 (PE-Cy7; BD Biolgend, San Diego, CA), CD8a (Alexa 647; BD Biolgend), CD25 (FITC; BD Biolgend), CD3 (PE; BD Biolegend), and CD161(PerCp; eBiosciences, San Diego, CA).

Techniques: Isolation, Staining

Journal: American Journal of Physiology - Renal Physiology

Article Title: IL-17 mediates neutrophil infiltration and renal fibrosis following recovery from ischemia reperfusion: compensatory role of natural killer cells in athymic rats

doi: 10.1152/ajprenal.00462.2016

Figure Lengend Snippet: Quantification of IL-17+ cells in response to IL-17 antagonism. Lymphocytes were obtained from postischemic euthymic and athymic rats fed high-salt diet (4.0% NaCl) and treated with vehicle or IL-17Rc antagonist. A: no. of total IL-17-producing cells isolated from rats treated with and without IL-17Rc. B–D: different sources of IL-17-producing cells: CD4+IL-17+ (B), CD8+IL-17+ (C), and CD4+CD161+IL-17+ (D). Data are means ±SE. *P < 0.05, IL-17Rc vs. vehicle using Student’s t-test (n = 3-5 animals/group).

Article Snippet: To evaluate T lymphocytes, the cells were stained with antibodies against rat CD4 (PE-Cy7; BD Biolgend, San Diego, CA), CD8a (Alexa 647; BD Biolgend), CD25 (FITC; BD Biolgend), CD3 (PE; BD Biolegend), and CD161(PerCp; eBiosciences, San Diego, CA).

Techniques: Isolation

Journal: Cell reports

Article Title: TCF1 and LEF1 Control Treg Competitive Survival and Tfr Development to Prevent Autoimmune Diseases

doi: 10.1016/j.celrep.2019.05.061

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rat monoclonal anti-CD4 (GK1.5) PE/Cy7 , TONBO , Cat#60-0041-U100; RRID:AB_2621828.

Techniques: Virus, Plasmid Preparation, Recombinant, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Diagnostic Assay, Software, Extraction, Gene Expression

Journal: Acta Biomaterialia

Article Title: Multipolymer microsphere delivery of SARS-CoV-2 antigens

doi: 10.1016/j.actbio.2022.12.043

Figure Lengend Snippet: Cellular immune response to MS-Ag administration in SD rats. SD rats (average body weight = 270 g) were each injected with a single intramuscular dose of 6 µg/g of body weight of chemically inactivated SARS-CoV-2 loaded MS (MS-Ag). From SD rats injected with MS-Ag, empty MS or saline, T-cell subsets of splenocytes were analyzed at 7 and 28 dpi by flow cytometry. (a-b) Percentage population of activated T-cells (CD4+CD69+ and CD8+CD69+), 28 dpi. (c-d) Percentage population of TCM - cells (CD4+CD62L+CD44+ and CD8+CD62L+CD44+), 28 dpi. Intracellular cytokine staining and flow cytometric analysis of CD4+ T-cell subset expressing (e) IFNγ at 7 dpi, (f) IFNγ at 28 dpi, (g) IL4 at 28 dpi, and (h) IL17 at 28 dpi. Statistical analysis was done by One-way ANOVA followed by Newman/Keul's post-hoc analysis for multiple comparisons, (*P < 0.05; **P < 0.01; ***P < 0.001). Abbreviations: MS, multipolymer microsphere; Ag, antigen; IFNγ, Interferon-gamma; IL, Interleukin; %, percentage of the parent population.

Article Snippet: For extracellular staining, 2 × 10 6 of spleen cells were first stained with LIVE/DEADTM Fixable Blue Dead Cell Stain Kit in PBS for 30 min. After saline washes, cells were then stained with CD3-PE (201412, Biolegend), CD4-BUV737 (741770, BD Biosciences), CD8a-BV650 (740514, BD Biosciences), CD62L-eFluor660 (50-0623-82), CD44-APC Cy7 (NB600-1317APCCY7, Novus Biologicals), CD69-AlexaFluor488 (bs-2499R-A488) antibodies to detect surface antigens.

Techniques: Injection, Tandem Mass Spectroscopy, Saline, Flow Cytometry, Staining, Expressing

Journal: Cancer Cell

Article Title: Cooperative Enhancer Activation by TLX1 and STAT5 Drives Development of NUP214-ABL1/TLX1-Positive T Cell Acute Lymphoblastic Leukemia

doi: 10.1016/j.ccell.2018.07.007

Figure Lengend Snippet:

Article Snippet: Rat monoclonal anti-CD4 (clone GK1.5) (PE Cy7) , Thermo Fischer Scientific , Cat# 25-0041-81, RRID: AB_469575.

Techniques: Recombinant, Infection, Transfection, Protease Inhibitor, Magnetic Beads, Lysis, Reverse Transcription, Purification, Cell Isolation, Western Blot, Polymer, Negative Control, Software