Splenocyte Immunophenotyping by Flow Cytometry

After isolation and re-stimulation, the splenocytes were collected, washed and immunophenotyped by using the following anti-mouse mAbs: CD3-eFluor 450, CD4-FITC (Thermo Fisher Scientific), and CD8a-APC-H7 (BD Biosciences). Appropriate isotype controls were used for comparison. After staining, the splenocytes were assessed by BD FACSAria™ II Flow Cytometer (BD Biosciences, San Jose, CA, USA). Dead cells were excluded by using LIVE/DEAD Fixable Dead Cell Stain (Invitrogen). Antigen specific CD4⁺ or CD8⁺ T cells were analyzed with FlowJo software (Tree Star Inc., Ashland, OR) [43 (link)]. All experiments were performed in triplicate.

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Bai X., Zhou Y., Yokota Y., Matsumoto Y., Zhai B., Maarouf N., Hayashi H., Carlson R., Zhang S., Sousa A., Sun B., Ghanbari H., Dong X, & Wands J.R. (2022). Adaptive antitumor immune response stimulated by bio-nanoparticle based vaccine and checkpoint blockade. Journal of Experimental & Clinical Cancer Research : CR, 41, 132.

Publication 2022

CD3-eFluor 450 CD4-FITC CD8a-APC-H7 BD FACSAria™ II Flow Cytometer LIVE/DEAD Fixable Dead Cell Stain

Antigen Cd8 t cells Cell Fitc Isolation Isotype Mabs Mouse Stain Tree

Corresponding Organization :

Other organizations : Brown University, Providence College, Rhode Island Hospital, First Affiliated Hospital of Harbin Medical University, Harbin Medical University

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Variable analysis

independent variables

Isolation and re-stimulation of splenocytes

dependent variables

Immunophenotype of splenocytes
Antigen-specific CD4+ or CD8+ T cells

control variables

Appropriate isotype controls

controls

Positive control: None specified
Negative control: Appropriate isotype controls

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