Journal: STAR Protocols

Article Title: High-dimensional functional phenotyping of preclinical human CAR T cells using mass cytometry

doi: 10.1016/j.xpro.2022.101174

Figure Lengend Snippet: Surface antibody mix

Article Snippet: Anti-Human CD25 (2A3)-169Tm (wd 1.00:50.00) , Fluidigm , Cat#3169003B.

Techniques:

Journal: STAR Protocols

Article Title: High-dimensional functional phenotyping of preclinical human CAR T cells using mass cytometry

doi: 10.1016/j.xpro.2022.101174

Figure Lengend Snippet: CAR T cell activation Representative histograms showing CD25 expression in non-CAR control and FMC63 CAR T cells in unstimulated (dark gray) and stimulated (light gray) conditions. Non-CAR control and CAR T cells were gated on live CD3+ and CD3+mCherry+ cells, respectively.

Article Snippet: Anti-Human CD25 (2A3)-169Tm (wd 1.00:50.00) , Fluidigm , Cat#3169003B.

Techniques: Activation Assay, Expressing, Control

Journal: STAR Protocols

Article Title: High-dimensional functional phenotyping of preclinical human CAR T cells using mass cytometry

doi: 10.1016/j.xpro.2022.101174

Figure Lengend Snippet:

Article Snippet: Anti-Human CD25 (2A3)-169Tm (wd 1.00:50.00) , Fluidigm , Cat#3169003B.

Techniques: Recombinant, Staining, Blocking Assay, Software, Cell Culture, Cell Counting, Cytometry, Irradiation, Microscopy

Journal: Frontiers in Immunology

Article Title: Human CD4 T Cells From Thymus and Cord Blood Are Convertible Into CD8 T Cells by IL-4

doi: 10.3389/fimmu.2022.834033

Figure Lengend Snippet: IL-4 induces the upregulation of CD8αβ on human CD4SP thymocytes and in peripheral CD4 + T cells. (A) CD4SP thymocytes, sorted as CD3 high CD4 + CD8 neg CD25 neg TCRγδ neg , were incubated with IL-2, IL-4 or IL-7 for 7 days and CD8α expression was assessed at the end of the culture. CD3 expression is presented as an heatmap overlay on initial populations. (B) Frequency of cytokine-induced double-positive (iDP) and CD8 single-positive (iCD8SP) thymocytes after 7 days of culture with IL-2, IL-4 or IL-7. Each dot represents a thymus. Statistics was only performed within the same subset (iDP or iCD8SP). (C) CD4 and CD8A mRNA expression in CD4SP and CD8SP thymocytes ex vivo and in sorted populations 7 days after IL-4 culture. tCD4SP: cytokine-treated CD4 single-positive thymocytes. (D) Expression of CD8β on tCD4SP, iDP and iCD8SP generated from incubation of CD4SP thymocytes with IL-4 for 7 days. (E) Expression of CD8α and CD8β on sorted naïve CD4 + T cells from cord, pediatric or adult blood and pediatric tonsil after a 7-day culture with IL-4 (left dot plots) and (F) quantification of iDP and iCD8SP cells generated from naïve and memory T cells (right panel; n=2/3). Results in graphs are presented as mean ± SD. *p < 0.05, ***p < 0.001, ****p < 0.0001.

Article Snippet: Anti-human monoclonal antibodies (mAbs) used were: CD3 (UCHT1), CD4 (RPA-T4), CD8α (RPA-T8), CD25 (2A3) and TCRγδ (B1.1) from eBioscience/Thermo Fisher Scientific or BD Biosciences.

Techniques: Incubation, Expressing, Ex Vivo, Generated

Journal: Frontiers in Immunology

Article Title: Human CD4 T Cells From Thymus and Cord Blood Are Convertible Into CD8 T Cells by IL-4

doi: 10.3389/fimmu.2022.834033

Figure Lengend Snippet: iCD8SP cells generated from CD4SP thymocytes in the presence of IL-4 are stable and diverse. (A) Proliferation of CD4SP thymocytes, sorted as CD3 high CD4 + CD8 neg CD25 neg TCRγδ neg cells, in response to IL-2, IL-4 and IL-7, as measured by Cell Trace Violet (CTV) dilution. Frequency of CTV neg cells is presented in each population, and data are representative of 3 experiments. (B) Proliferation of tCD4SP, iDP and iCD8SP thymocytes in response to IL-4, as measured by CTV dilution or Ki67 frequency (graph: n=5). (C) Stability of IL-4-induced CD8 expression, as assessed by sorting iDP and iCD8SP populations on day 7 of IL-4 culture and either maintaining cells in IL-4 or switching them to IL-2 for 7 more days. (D) CD8 induction in CD4 + CD8 neg (tCD4SP) cells sorted 7 days after IL-4 culture of CD4SP thymocytes and cultured in the presence of IL-4 for 7 additional days. (E) Bcl-2 median fluorescence intensity (MFI) of different populations before and after exposure to IL-4 or IL-7. (F) TCR Vβ repertoire of CD8SP, CD4SP and IL-4-induced iCD8SP thymocytes (n=3). Results in graphs are presented as mean ± SD.

Article Snippet: Anti-human monoclonal antibodies (mAbs) used were: CD3 (UCHT1), CD4 (RPA-T4), CD8α (RPA-T8), CD25 (2A3) and TCRγδ (B1.1) from eBioscience/Thermo Fisher Scientific or BD Biosciences.

Techniques: Generated, Expressing, Cell Culture, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Bone Marrow Mast Cell Antibody-Targetable Cell Surface Protein Expression Profiles in Systemic Mastocytosis

doi: 10.3390/ijms20030552

Figure Lengend Snippet: List of fluorochrome-conjugated monoclonal antibodies used in this study to characterize BMMCs from SM patients and normal/reactive BM.

Article Snippet: CD25 , 2A3 , PE , BD Biosciences * , Interleukin-2 receptor, subunit α.

Techniques: