4.4 Article

Analysis of Granulocyte-Macrophage Colony-Stimulating Factor-Producing T Helper Cells in a Mouse Model of Contact Hypersensitivity

Journal

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 181, Pages -

Publisher

JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/63755

Keywords

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Funding

  1. National Natural Science Foundation of China [81602763, 81803142, 82003347]
  2. Excellent Researcher Program of China Postdoctoral Science Foundation [2017T100700]
  3. Regular Researcher Program of China Postdoctoral Science Foundation [2016M592673]

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This study induced a novel T cell subset called Th-GM cells in a mouse CHS model and found that they were mainly expanded at the site of injury and draining lymph nodes. This method can be used to further investigate the biology of Th-GM cells and explore therapeutic strategies centered on GM-CSF in various conditions.
Parallel to traditional Th1/Th2/Th17/Treg lineages, granulocyte-macrophage colony-stimulating factor-producing T helper (Th-GM) cells have been identified as a distinct subset of T helper cells (GM-CSF+ IFN-gamma(-) IL-17A(-) IL-22(-) effector CD4(+) T cells) in human and mice. Contact hypersensitivity (CHS) is considered an excellent animal model for allergic contact dermatitis (ACD) in human, manifesting an intact T cell-mediated immune response. To provide a standardized and comprehensive assay to analyze the Th-GM cell subset in the T cell-dependent immune response in vivo, a murine CHS model was induced by sensitization/challenge with a reactive, low-molecular-weight, organic hapten, 2,4-dinitrofluorobenzene (DNFB). The Th-GM subset in effector CD4(+) T cells generated upon immunization with the hapten was analyzed by flow cytometry. We found that Th-GM was mainly expanded in lesions and draining lymph nodes in the DNFB-induced CHS mouse model. This method can be applied to further study the biology of Th-GM cells and pharmacological research of therapeutic strategies centered on GM-CSF in various conditions, such as ACD.

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