Activation of mast cells by streptolysin O and lipopolysaccharide.
This chapter provides protocols to measure the reversible permeabilization of mast cells by streptolysin O (SLO) and to follow SLO-induced activation of mast cells by monitoring degranulation, activation of mitogen-activated protein kinases, and production of tumor necrosis factor-alpha. A method that uses SLO to deliver molecules into the cytosol of living cells also is described. Furthermore, we outline a procedure to measure the activation of nuclear factor-kappaB by lipopolysaccharide and ionomycin using transfection of mast cells with reporter genes by electroporation. These protocols should be widely applicable in mast cell research.
Stassen
Michael
M
Institute of Immunology, Johannes Gutenberg-University, Mainz, Germany.
Valeva
Angela
A
Walev
Iwan
I
Schmitt
Edgar
E
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins / Q000276:immunology; D001854:Bone Marrow Cells / Q000276:immunology; D002463:Cell Membrane Permeability; D018274:Electroporation / Q000379:methods; D004789:Enzyme Activation; D015759:Ionomycin / Q000378:metabolism; D007476:Ionophores / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology*; D008407:Mast Cells / Q000276:immunology*; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D013301:Streptolysins / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
D001426:Bacterial Proteins; D007476:Ionophores; D008070:Lipopolysaccharides; D016328:NF-kappa B; D013301:Streptolysins; D014409:Tumor Necrosis Factor-alpha; C025964:streptolysin O; D015759:Ionomycin; D000255:Adenosine Triphosphate; D020928:Mitogen-Activated Protein Kinases; D001619:beta-N-Acetylhexosaminidases