FIGURE 6.

hCLEC9A can induce pro-inflammatory cytokine production and can recruit and signal via Syk kinase. A, quantitation of zymosan (zym) induced TNF production by transduced RAW264.7 macrophages, in the presence or absence of soluble β-glucan (β-glu), as indicated, demonstrating the ability of the chimeric receptor to induce proinflammatory cytokine production. Shown are the mean ± S.D. of one representative experiment of three. B, Western blotting of immunoprecipitates using phosphorylated (pY) and unphosphorylated peptides (Y) corresponding to the cytoplasmic tail of hCLEC9A or Dectin-1 from RAW264.7 cell lysates. Blots were probed with anti-phosphotyrosine (αPY), anti-Syk, and anti-Lyn, as indicated. C, IL-2 production following zymosan stimulation of Syk-deficient (Syk-) and Syk-sufficient (Syk+) cells, transduced with the chimeric receptor or vector-only control, as indicated, showing that cytokine production induced by the chimeric receptor in response to zymosan requires Syk. The data shown are the mean ± S.D. and are representative of three independent experiments. ^*, p < 0.05 versus control (Student's t test).