Liver Leukocyte Isolation and Flow Cytometry

Leukocytes were isolated from the liver in the way as described previously^{40 (link)}. For flow cytometry, cells were stained for surface antigens with anti-NK1.1, anti-CD3, anti-CD4, anti-CD8α, anti-CD19, anti-CD44, anti-CD62L (BioLegend, San Diego, CA), or/and for intracellular antigens with anti-IFN-γ, anti-IL-4, anti-IL-17A, anti-TNF-α and anti-IL-10 (BioLegend). IgG isotype controls (Biolegend) were used in parallel. To detect Treg cells, cells were stained with anti-NK1.1, anti-CD3, anti-CD4, anti-CD25 at 4 °C for 30 minutes; and incubated with anti-mouse Foxp3 antibodies according to the manufacturer’s instructions (eBioscience). Methods were the sameas those described previously^{9 (link)}.

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Li Z., Zhang C., Li L., Bi X., Li L., Yang S., Zhang N., Wang H., Yang N., Abulizi A., Aini A., Lin R., Vuitton D.A, & Wen H. (2019). The local immune response during Echinococcus granulosus growth in a quantitative hepatic experimental model. Scientific Reports, 9, 19612.

Publication 2019

anti-NK1.1 anti-CD3 anti-CD4 anti-CD8α anti-CD19 anti-CD44 anti-CD62L anti-IFN-γ anti-IL-4 anti-IL-17A anti-TNF-α anti-IL-10 IgG isotype controls anti-CD25 anti-mouse Foxp3 antibodies

Anti antibodies Anti cd3 Antigens Cd25 Cd44 Cd62l Cells Flow cytometry Ifn γ Il 10 Il 17a Intracellular Isotype Leukocytes Liver Mouse Surface antigens Tnf α Treg cells

Corresponding Organization :

Other organizations : Xinjiang Medical University, First Affiliated Hospital of Xinjiang Medical University

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Variable analysis

independent variables

Isolation method for leukocytes from liver

dependent variables

Surface antigen expression (NK1.1, CD3, CD4, CD8α, CD19, CD44, CD62L)
Intracellular antigen expression (IFN-γ, IL-4, IL-17A, TNF-α, IL-10)
Regulatory T cell (Treg) identification (CD3, CD4, CD25, FoxP3)

control variables

IgG isotype controls for surface and intracellular antigen staining

controls

Positive control: Not specified
Negative control: IgG isotype controls for surface and intracellular antigen staining

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