T Cell Depletion and Flow Cytometry

At 3 days and 1 day before viral challenge, all cohorts of mice were treated intraperitoneally with 500 μg each of both anti-CD4 (clone GK1.5) and anti-CD8a (YTS 169.4) (BioXcell)52 (link)53 (link). Blood samples from each mouse were then assessed for the presence of CD4 and CD8 T cells by flow cytometry. Cells were fluorescently labelled with anti-CD4 (clone RM4-5, ebioscience) and anti-CD8 (clone 53-6.7, BD) at a dilution of 1:200 in FACS buffer (1% fetal bovine serum in PBS). Cells were then analysed using the BD FACSCalibur and flow plots created in FlowJo 8.7.

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Bohannon C., Powers R., Satyabhama L., Cui A., Tipton C., Michaeli M., Skountzou I., Mittler R.S., Kleinstein S.H., Mehr R., Lee F.E., Sanz I, & Jacob J. (2016). Long-lived antigen-induced IgM plasma cells demonstrate somatic mutations and contribute to long-term protection. Nature Communications, 7, 11826.

Publication 2016

anti-CD4 (clone GK1.5) anti-CD8a (YTS 169.4) anti-CD4 (clone RM4-5, anti-CD8 (clone 53-6.7, BD FACSCalibur

Blood Buffer Cd8 t cells Cells Clone Dilution Fetal bovine serum Flow cytometry Mouse

Corresponding Organization :

Other organizations : Emory University, Yale University, Bar-Ilan University

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Variable analysis

independent variables

Treatment with anti-CD4 (clone GK1.5) and anti-CD8a (YTS 169.4) antibodies

dependent variables

Presence of CD4 and CD8 T cells in blood samples

control variables

Timing of antibody treatment (3 days and 1 day before viral challenge)
Dose of antibodies (500 μg each)
Fluorescent labeling of cells (anti-CD4 and anti-CD8 at 1:200 dilution in FACS buffer)
Flow cytometry analysis (BD FACSCalibur)

controls

No positive control is explicitly mentioned.
No negative control is explicitly mentioned.

Annotations

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