title: Isolation and Activation of Murine Lymphocytes.
abstract: B and T cells, with their extremely diverse antigen-receptor repertoires, have the ability to mount specific immune responses against almost any invading pathogen1,2. Understandably, such intricate abilities are controlled by a large number of molecules involved in various cellular processes to ensure timely and spatially regulated immune responses3. Here, we describe experimental procedures that allow rapid isolation of highly purified murine lymphocytes using magnetic cell sorting technology. The resulting purified lymphocytes can then be subjected to various in vitro or in vivo functional assays, such as the determination of lymphocyte signaling capacity upon stimulation by immunoblotting4and the investigation of proliferative abilities by3H-thymidine incorporation or carboxyfluorescein diacetate succinimidyl ester (CFSE) labeling5-7. In addition to comparing the functional capacities of control and genetically modified lymphocytes, we can also determine the T cell stimulatory capacity of antigen-presenting cells (APCs) in vivo, as shown in our representative results using transplanted CFSE-labeled OT-I T cells.
authors.affiliation: School of Biological Sciences, College of Science, Nanyang Technological University.
authors.forename: Jun Feng
authors.initials: JF
authors.lastname: Lim
authors.affiliation: School of Biological Sciences, College of Science, Nanyang Technological University.
authors.forename: Heidi
authors.initials: H
authors.lastname: Berger
authors.affiliation: School of Biological Sciences, College of Science, Nanyang Technological University; ihsu@ntu.edu.sg.
authors.forename: I-Hsin
authors.initials: IH
authors.lastname: Su
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry; D005452:Fluoresceins; D008213:Lymphocyte Activation; D008214:Lymphocytes; D008280:Magnetics; D051379:Mice; D013388:Succinimides; D013601:T-Lymphocytes
chemical_list: D005452:Fluoresceins; D013388:Succinimides