Immune Cell Profiling in Lungs

To analyze immune lung infiltration, lung suspensions were stained as previously described [20 (link)] using the following directly conjugated antibodies: CD3e FITC (Miltenyi, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany, clone 145-2C11); CD11b PE (BD Biosciences, San Jose, CA USA, clone M1/70); CD11c PECY7 (Thermo Fisher Scientific-eBiosciences, clone N418); CD45 APCeFluor780 (Thermo Fisher Scientific-eBioscience, clone 30-F11); CD49b PE (Miltenyi, clone DX5); CD69 APC (Miltenyi, clone H1.2F3); CD103 APC (Miltenyi, clone REA789); B220 PERCPVio700 (Miltenyi, clone RA3-6B2); and mPDCA-1 PE (Miltenyi, clone JF05-1C2.4.1). A purified rat anti-mouse CD16/CD32 MAb (Thermo Fisher Scientific-eBioscience, clone 93) was used to block nonspecific binding to mouse Fc receptors. The cells were analyzed using a FACSCanto flow cytometer (BD Biosciences) and FlowJo software (TreeStar). All analyses were performed using gating on CD45+ live cells after doublet exclusion.

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Le Noci V., Sommariva M., Bianchi F., Triulzi T., Tagliabue E., Balsari A, & Sfondrini L. (2019). Local Administration of Caloric Restriction Mimetics to Promote the Immune Control of Lung Metastases. Journal of Immunology Research, 2019, 2015892.

Publication 2019

CD3e FITC CD11b PE CD49b PE CD69 APC FACSCanto

Antibodies Block Cd103 Cd11b Cells Clone Fc receptors Fitc Lung Mouse

Corresponding Organization : University of Milan

Other organizations : Fondazione IRCCS Istituto Nazionale dei Tumori

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Variable analysis

independent variables

Directly conjugated antibodies used for staining: CD3e FITC, CD11b PE, CD11c PECY7, CD45 APCeFluor780, CD49b PE, CD69 APC, CD103 APC, B220 PERCPVio700, and mPDCA-1 PE

dependent variables

Immune lung infiltration

control variables

Purified rat anti-mouse CD16/CD32 MAb was used to block nonspecific binding to mouse Fc receptors
All analyses were performed using gating on CD45+ live cells after doublet exclusion

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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