Augmentation of Staphylococcal α-Toxin Signaling by the Epidermal Platelet-Activating Factor Receptor Jeffrey B. Travers, Donald Y.M. Leung, Christopher.

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Presentation transcript:

Augmentation of Staphylococcal α-Toxin Signaling by the Epidermal Platelet-Activating Factor Receptor Jeffrey B. Travers, Donald Y.M. Leung, Christopher Johnson, Patrick Schlievert, Mariangela Marques, Jason Cosgrove, Keith L. Clay Journal of Investigative Dermatology Volume 120, Issue 5, Pages 789-794 (May 2003) DOI: 10.1046/j.1523-1747.2003.12149.x Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 The effect of staphylococcal α-toxin on PAF production in HaCaT keratinocytes. (A) HaCaT cells were incubated with 2.5 μg α-toxin per ml, and the 1-hexadecyl and 1-palmitoyl sn-2 acetyl GPC species were measured at various times as described in Materials and Methods. (B) HaCaT cells were incubated with the indicated dosages of α-toxin for 5 min, and the 1-hexadecyl and 1-palmitoyl sn-2 acetyl GPC species were measured. The values are expressed as mean±SEM of at least three to four separate experiments using duplicate samples. Journal of Investigative Dermatology 2003 120, 789-794DOI: (10.1046/j.1523-1747.2003.12149.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 The effect of staphylococcal α-toxin on arachidonic acid release in HaCaT keratinocytes. HaCaT cells were incubated with 2.5 μg α-toxin per ml, and supernatant-associated arachidonic acid was measured at various times as described in Materials and Methods. Each value is the mean±SEM of at least three separate experiments using duplicate samples. Journal of Investigative Dermatology 2003 120, 789-794DOI: (10.1046/j.1523-1747.2003.12149.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 The effect of PAF-R antagonists on α-toxin-induced arachidonic acid release in HaCaT keratinocytes. HaCaT cells were preincubated with either 10 μM or 25 μM of the PAF-R antagonists WEB 2086 or A-85789 for 30 min before addition of 2.5 μg α-toxin per ml, and supernatant-associated arachidonic acid was measured after 60 min The values are mean±SD of duplicate experiments from a single experiment from three experiments with similar results. *Pretreatment of HaCaT cells with 25 μM WEB 2086 and 10 μM and 25 μM A-85789 resulted in a statistically significant (p<0.05) decrease in α-toxin-induced arachidonic acid release. Journal of Investigative Dermatology 2003 120, 789-794DOI: (10.1046/j.1523-1747.2003.12149.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Effect of overexpression of PAF-AHII on α-toxin-induced arachidonic acid release in HaCaT keratinocytes. HaCaT cells transduced with PAF-AHII (HaCaTMPAF-AHII) or control retrovirus (HaCaTM) were treated with 2.5 μg α-toxin per ml, and supernatant-associated arachidonic acid was measured after 60 min. The values are mean±SD of duplicate samples from a single experiment from three experiments with similar results. *Treatment of HaCaTMPAF-AHII cells with α-toxin resulted in a statistically significant (p<0.05) decrease in arachidonic acid release over HaCaTM cells. Journal of Investigative Dermatology 2003 120, 789-794DOI: (10.1046/j.1523-1747.2003.12149.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Effect of PAF-R expression on a-toxin-induced arachidonic acid release in KB cells. (A) KB cells transduced with PAF-R (KBP) or control retrovirus (KBM) were treated with 2.5 μg α-toxin per ml for various times, and supernatant-associated arachidonic acid was measured at various times. (B) KBM or KBP cells were preincubated with 25 μM of PAF-R antagonists WEB 2086 or A-85789 for 30 min before treatment with 2.5 μg α-toxin per ml, and supernatant-associated arachidonic acid was measured after 60 min. The values are mean±SD of duplicate samples from a single experiment from three experiments with similar results. Journal of Investigative Dermatology 2003 120, 789-794DOI: (10.1046/j.1523-1747.2003.12149.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Effect of α-toxin on intracellular calcium levels in KB cells. KBM or KBP cells were loaded with the calcium-sensitive dye Indo-1 and treated with 100 nM PAF, 1 μM endothelin-1 (ET-1), or 2.5 μg α-toxin per ml (α-T). Intracellular Ca2+ levels were calculated at the various times from fluorescent values as previously described (Pei et al, 1998). Journal of Investigative Dermatology 2003 120, 789-794DOI: (10.1046/j.1523-1747.2003.12149.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions