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. 2016 Dec 12;213(13):2871–2883. doi: 10.1084/jem.20160597

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© 2016 Kretzer et al.

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Figure 4. — CD8α⁺ DCs from Rab43^Δ/Δ mice are defective in cross-presentation of cell-associated and soluble antigens. (A and B) Sorted CD8α⁺ (A) or CD8α⁻ (B) DCs from WT (black) or Rab43^Δ/Δ (red) B6 mice were cultured for 3 d with CFSE-labeled OT-I T cells and different doses of HKLM-OVA and assayed for OT-I proliferation and activation (CFSE⁻CD44⁺). (C and D) Sorted CD8α⁺ (C) or CD8α⁻ (D) DCs from WT (black) or Rab43^Δ/Δ (red) mice were cultured and analyzed as in A with various doses of soluble OVA as antigen. (E) Sorted CD8α⁺ (continuous lines) or CD8α⁻ (dashed lines) DCs from WT (black) or Rab43^Δ/Δ (red) mice were cultured and analyzed as in A with the indicated amounts of SIINFEKL peptide as antigen. (F) Rab43^f/f-CD11c^Cre− (black) or Rab43^f/f-CD11c^Cre+ (red) mice were injected with the indicated numbers of PBS (OVA⁻)- or OVA (OVA⁺)-loaded irradiated MHCI TKO splenocytes and analyzed 8 d later for the number of K^b SIINFEKL tetramer⁺ (tet⁺) T cells that were CD44⁺CD62L⁻. Each dot represents cells obtained from one mouse. (G) Sorted CD8α⁺ (solid) or CD8α⁻ (dashed) DCs from WT (black) or Rab43^Δ/Δ (red) mice were osmotically loaded with OVA, cultured, and analyzed as in A. (H) Sorted CD8α⁺ (solid) or CD8α⁻ (dashed) DCs from WT (black) or Rab43^Δ/Δ (red) mice were cultured for 3 d with CFSE-labeled OT-II T cells and irradiated splenocytes osmotically loaded with OVA. T cells were assayed for proliferation and activation (CFSE⁻CD44⁺). All data are displayed as mean ± SEM from at least two independent experiments. Statistics were analyzed using two-way ANOVA. ***, P < 0.001; ****, P < 0.0001.