Journal:

Article Title: Induction of Specific Cytotoxic Lymphocytes in Mice Vaccinated with Brucella abortus RB51

doi: 10.1128/IAI.69.9.5502-5508.2001

Figure Lengend Snippet: Different roles of CD4+ and CD8+ T cells in the cytotoxic lysis and release of IFN-γ. The two T-cell populations were isolated by the MACS magnetic kit and showed more than 92% purity by flow cytometry. The individual T-cell populations were cocultured at various concentrations with either strain RB51-infected or normal J774.A1 cells. The cytotoxic activity (A) and the amount of IFN-γ released into the supernatants (B) were measured. The data are means for triplicate estimations, and standard deviations did not exceed 20% of the means. Cocultures of CD4+ T cells with noninfected target macrophages (solid squares) and RB51-pulsed target macrophages (solid triangles) and CD8+ T cells with noninfected target cells (solid diamonds) and RB51-pulsed target cells (solid circles) were tested.

Article Snippet: Briefly, live T cells isolated by Histopaque column purification were incubated with MACS magnetic MicroBeads to which monoclonal antibodies against CD4 molecule (clone GK1.5; isotype, rat IgG2b) or CD8 molecule (clone 53-6.7; isotype, rat IgG2a) had been coupled (Miltenyi Biotec, Auburn, Calif.) at the concentration of 10 μl of MicroBeads per 10 7 total cells for 15 min in a refrigerator at 4°C (see Fig. ).

Techniques: Lysis, Isolation, Flow Cytometry, Infection, Activity Assay

Journal:

Article Title: Induction of Specific Cytotoxic Lymphocytes in Mice Vaccinated with Brucella abortus RB51

doi: 10.1128/IAI.69.9.5502-5508.2001

Figure Lengend Snippet: Specific phenotype analysis of effector cells by flow cytometry a

Article Snippet: Briefly, live T cells isolated by Histopaque column purification were incubated with MACS magnetic MicroBeads to which monoclonal antibodies against CD4 molecule (clone GK1.5; isotype, rat IgG2b) or CD8 molecule (clone 53-6.7; isotype, rat IgG2a) had been coupled (Miltenyi Biotec, Auburn, Calif.) at the concentration of 10 μl of MicroBeads per 10 7 total cells for 15 min in a refrigerator at 4°C (see Fig. ).

Techniques: Flow Cytometry

Journal: Nature Communications

Article Title: Fasting mimicking diet in mice delays cancer growth and reduces immunotherapy-associated cardiovascular and systemic side effects

doi: 10.1038/s41467-023-41066-3

Figure Lengend Snippet:

Article Snippet: Anti-mouse CD4, VioBright FITC (REA604) , Miltenyi Biotec , 130-118-692.

Techniques: In Vivo

Journal: Cell reports

Article Title: PARP-inhibition reprograms macrophages toward an anti-tumor phenotype

doi: 10.1016/j.celrep.2022.111462

Figure Lengend Snippet:

Article Snippet: Anti-Mouse CD4 (RM4-5)-145Nd , Fluidigm , Cat# 3145002B; RRID:AB_2687832.

Techniques: Control, Recombinant, Liposomes, Labeling, Quantitation Assay, Staining, RNA Sequencing, Mass Spectrometry, Knock-Out, Software

Journal:

Article Title: Neuroprotective autoimmunity: Naturally occurring CD4 + CD25 + regulatory T cells suppress the ability to withstand injury to the central nervous system

doi: 10.1073/pnas.232565399

Figure Lengend Snippet: Depletion of naturally occurring regulatory CD4+CD25+ T cells in BALB/c mice improves neuronal survival after optic nerve crush injury. (A) BALB/c mice were thymectomized (ThyX) 3 days after birth to deplete their regulatory T cells and were subjected as adults to severe unilateral crush injury inflicted on the intraorbital portion of the optic nerve. Surviving neurons were labeled by the application, 3 days before injury, of the neurotracer dye FluoroGold. Significantly more neurons survived in the thymectomized mice than in control (nonthymectomized) age-matched mice (n = 7–8 in each group; P < 0.001; Student's t test). (B) Neuronal survival after optic nerve crush injury in BALB/c nu/nu mice (devoid of T cells) was worse than in wild-type mice of matched background. Endogenous neuroprotection in the nude mice was restored by injection of 5 × 107 wild-type splenocytes. Injection of splenocytes depleted of regulatory CD4+CD25+ T cells increased neuronal survival in these mice beyond even that seen in the wild type (n = 5–6 in each group; P values between different groups, obtained by two-tailed Student's t test, are indicated by asterisks above the graph bars; *, P < 0.05; **, P < 0.01; ***, P < 0.001).

Article Snippet: Mouse anti-rat CD4 antibodies conjugated to phycoerythrin (PE) and mouse anti-rat CD25 antibodies conjugated to fluorescein isothiocyanate were purchased from Serotec.

Techniques: Labeling, Injection, Two Tailed Test

Journal:

Article Title: Neuroprotective autoimmunity: Naturally occurring CD4 + CD25 + regulatory T cells suppress the ability to withstand injury to the central nervous system

doi: 10.1073/pnas.232565399

Figure Lengend Snippet: Naturally occurring regulatory CD4+CD25+ T cells diminish spontaneous neuroprotection. BALB/c mice, endowed with the spontaneous ability to evoke a protective beneficial autoimmune response, were injected with 2 × 106 purified activated regulatory CD4+CD25+ T cells. Control mice were injected with 2 × 106 purified activated effector cells (CD4+CD25−) or with PBS. Injection of regulatory T cells had an adverse effect on neuronal survival (more neurons underwent secondary degeneration). The results shown are of one representative experiment of three independent experiments; n = 5–6 mice in each group (P < 0.01 and 0.05; respectively).

Article Snippet: Mouse anti-rat CD4 antibodies conjugated to phycoerythrin (PE) and mouse anti-rat CD25 antibodies conjugated to fluorescein isothiocyanate were purchased from Serotec.

Techniques: Injection, Purification

Journal: Animals : an Open Access Journal from MDPI

Article Title: Evaluation of a Novel Precision Biotic on Enterohepatic Health Markers and Growth Performance of Broiler Chickens under Enteric Challenge

doi: 10.3390/ani12192502

Figure Lengend Snippet: Target phenotypes used in analysis of intestinal T cells.

Article Snippet: CD4 , T-helper cells , MCA2164F Biorad.

Techniques: Marker

Journal: Frontiers in Immunology

Article Title: Characterisation of the bovine C-type lectin receptor Mincle and potential evidence for an endogenous ligand

doi: 10.3389/fimmu.2023.1189587

Figure Lengend Snippet: Identification of a potential boMincle ligand expressed on bovine CD4 + T cells. To identify potential self-derived boMincle ligands, CFSE-labelled CD4 + T cells were incubated with pUNO1-boMincle-transfected CHO cells. CHO cells were either untreated or pre-incubated with anti-boMincle antibody AbD31662. CHO cells transfected with empty pUNO1 and pUNO1-boDC-SIGN plasmid DNA served as negative and positive control, respectively. The number of CHO cell-bound CD4 + T cells was analysed by fluorescence microscopy. The results of two individual experiments, for which six randomly selected fields of view per condition were counted, are summarised as truncated violin plot. The median for each condition is shown as continuous line and the 75% and 25% quartiles by dotted lines (p = 0.0332 (*), p = 0.0021 (**), p = 0.0002 (***)).

Article Snippet: Bovine CD4 + T cells were isolated from PBMCs by magnetic-activated cell sorting (MACS) using a mouse anti-bovine CD4 antibody (clone CC8, Bio-Rad), anti-mouse IgG MicroBeads (Miltenyi Biotec), and LS columns (Miltenyi Biotec), according to the manufacturer’s instructions.

Techniques: Derivative Assay, Incubation, Transfection, Plasmid Preparation, Positive Control, Fluorescence, Microscopy