Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Cutting edge: T cells trigger CD40-dependent platelet activation and granular RANTES release: a novel pathway for immune response amplification.

doi: 10.4049/jimmunol.172.4.2011

Figure Lengend Snippet: FIGURE 3. CD40-dependent RANTES release by platelets. RANTES re- lease by soluble and membrane-bound CD40-stimulated platelets. Platelets were stimulated either with sCD40L or paraformaldehyde-fixed D1.1 cells (1: 10), and RANTES content in the supernatants was measured by ELISA. Data are mean SEM of three separate experiments. , p 0.01 for sCD40L- and D1.1 cell-stimulated platelets vs blocking Ab treatment.

Article Snippet: RANTES ELISA kits and human recombinant RANTES were purchased from R&D Systems (Minneapolis, MN) and phospho-p38 and phospho-extracellular signalregulated kinase (ERK) Abs from Cell Signaling (Beverly, MA).

Techniques: Membrane, Enzyme-linked Immunosorbent Assay, Blocking Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Cutting edge: T cells trigger CD40-dependent platelet activation and granular RANTES release: a novel pathway for immune response amplification.

doi: 10.4049/jimmunol.172.4.2011

Figure Lengend Snippet: FIGURE 4. Involvement of p38 MAP kinase in platelet CD40 signaling. A, Immunoblotting of CD40-induced p38 phosphorylation in sCD40L-stimulated platelets. Platelets were stimulated with sCD40L and then lysed at the indicated time points. Immunoblotting was performed with Abs specific for phosphorylated p38 and ERK. This figure is representative of three separate experiments. B, Inhi- bition of CD40-dependent platelet activation by SB203580. Platelets were acti- vated with sCD40L with or without pretreatment with the p38 MAP kinase inhib- itor SB203580 and then submitted to flow cytometric analysis for P-selectin expression. The black curve represents the background signal from the isotype con- trol. The presence of DMSO, used for SB203580 dilution, had no effect on P- selectin expression. Data represent the mean SEM of three separate experiments. , p 0.01 for inhibitor-treated compared with sCD40L-stimulated platelets. C, Inhibition of CD40-dependent RANTES release by SB203580. Platelets were stimulated with sCD40L in the presence and absence of SB203580, and RANTES content in the supernatants was measured by ELISA. Data are mean SEM of three separate experiments. , p 0.01 for inhibitor-treated compared with sCD40L-stimulated platelets.

Article Snippet: RANTES ELISA kits and human recombinant RANTES were purchased from R&D Systems (Minneapolis, MN) and phospho-p38 and phospho-extracellular signalregulated kinase (ERK) Abs from Cell Signaling (Beverly, MA).

Techniques: Western Blot, Phospho-proteomics, Activation Assay, Inhibition, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Cutting edge: T cells trigger CD40-dependent platelet activation and granular RANTES release: a novel pathway for immune response amplification.

doi: 10.4049/jimmunol.172.4.2011

Figure Lengend Snippet: FIGURE 5. Platelet-derived RANTES mediates increased T cell recruitment. A, Detection of platelet-derived and recombinant RANTES on the surface of HIMEC by confocal microscopy. Confluent HIMEC monolayers were exposed to degranulated platelet-derived or recombinant (r) RANTES and fluorescently labeled for detection of RANTES (Alexa 488 secondary Ab, green) and nuclei (DAPI, blue). This figure is representative of four separate experiments. B, Platelet-derived RANTES-mediated T cell adhesion to HIMEC and inhibition by met-RANTES. HIMEC monolayers were left untreated (baseline) or exposed to degranulated platelet-derived or recombinant RANTES. MOLT4 cells, preincubated or not with met-RANTES, were added to the HIMEC monolayers. The number of adherent cells in each experimental condition was expressed as mean SEM of four separate experiments. , p 0.05 for met-RANTES-pretreated compared with untreated MOLT4 cells.

Article Snippet: RANTES ELISA kits and human recombinant RANTES were purchased from R&D Systems (Minneapolis, MN) and phospho-p38 and phospho-extracellular signalregulated kinase (ERK) Abs from Cell Signaling (Beverly, MA).

Techniques: Derivative Assay, Recombinant, Confocal Microscopy, Labeling, Inhibition