_Dr. Miller's lecture_chemokine_lecture_2

_Dr. Miller's lecture_chemokine_lecture_2 - Experimental...

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Pertussis toxin specifically inhibits signal transduction mediated through G α i (G protein)-linked receptors Experimental evidence for the role of chemokines in leukocyte trafficking
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Evidence for chemokine-mediated Evidence for chemokine-mediated activation of leukocyte integrins ( activation of leukocyte integrins ( in vivo) in vivo)
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Label lymphocytes with fluorochrome and treat with pertussis toxin Inject lymphocytes into mouse tail vein Isolate lymphocytes from mouse lymph nodes Treatment of leukocytes with pertussis toxin inhibits recruitment to sites of inflammation in vivo (Bargatze and Butcher, 1993)
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Isolate and exteriorize segments of small intestine and Peyer’s patches Visualize and count labeled lymphocytes within high endothelial venules via fluorescence microscopy
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In situ video microscopic analysis of lymphocyte-high endothelial cell interactions
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Evidence for chemokine-mediated Evidence for chemokine-mediated activation of leukocyte integrins ( activation of leukocyte integrins ( in vitro) in vitro)
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ICAM Static adhesion assay Treatment of leukocytes with pertussis toxin in vitro inhibits integrin-dependent triggering and arrest (Campbell, et. al. 1998)
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Pertussis toxin inhibits chemokine-triggered lymphocyte adhesion to ICAM-1
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Adhesion of CD4+ T cells to ICAM-1 in response to chemokines
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Adhesion of memory and naive CD4+ T cell subsets to ICAM-1 in response to chemokines
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Flow chamber assay
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chemokines
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_Dr. Miller's lecture_chemokine_lecture_2 - Experimental...

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