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. Author manuscript; available in PMC: 2008 Apr 1.
Published in final edited form as: Immunity. 2007 Mar 29;26(4):503–517. doi: 10.1016/j.immuni.2007.03.006

Figure 4. Local priming of NK cells requires interaction with DC and requires the recruitment of NK cells to lymph nodes.

Figure 4

(A and B) One day after DT injection, B6 mice (solid symbols) or CD11c DTR tg mice (open symbols) were injected subcutaneously (s.c.) into one footpad with the indicated TLR ligand. NK cells were purified from the indicated organs and IFN-γ production (A) and cytotoxicity (B) was determined in response to RMA-S/H60 cells 18-24h later. Similar results were obtained with YAC-1 and RMA-S target cells (data not shown). Error bars display s.d. (n=3-4) and results are representative of three separate experiments.

(C and D) Blocking entry of NK cells into LN results in dramatically decreased numbers of effector NK cells in peripheral tissues. B6 mice were injected with control Ig (Ctrl. Ig; solid symbols) or an anti-CD62L antibody (open symbols) blocking entry of NK cells into the LN. One day later, mice were injected with the indicated TLR ligands. Cytotoxicity (C) and IFN-γ production (D) of NK cells purified from the indicated tissues in response to RMA-S/H60 cells was determined 18-24h after local TLR stimulation. Similar results were obtained with RMA-S and YAC-1 targets (data not shown). Error bars display s.d. (n=3-4) and results are representative of three separate experiments.

(E and F) Circulating naïve NK cells need to enter the draining LN for priming. B6 mice (CD45.2+) were injected i.p. with control Ig (solid symbols) or an anti-CD62L antibody (open symbols). Two hours later, 3×106 purified NK cells (CD45.1+) were injected i.v. followed by TLR stimulation 12h later. IFN-γ production (E) and granzyme B expression (F) of transferred CD45.1+ NK cells isolated from the indicated tissues in response to RMA-S/H60 cells was determined by intracellular cytokine staining and electronic gating on NK cells 18-24h after local TLR stimulation. Similar results were obtained with RMA-S and YAC-1 targets (data not shown). Error bars display s.d. (n=3-4) and results are representative of two separate experiments. nd, not done.