FIGURE 4.

JAK2 V617F^hom DC migrate toward CCL19 chemokine and induce polarization of CD8⁺ T cells toward CD8⁺ CD103⁺ tissue resident T cells. (A) DC obtained from JAK2 V617F^hom iPS cells (patient PV2) and the respective iPS cells without mutation (JAK2) migrate toward CCL19 chemokine. CD1c⁺ HLA-DR⁺ cells of day 8–9 of DC differentiation were treated with LPS (1 μg/ml, 24 h) or left untreated and subjected to CCL19 chemotaxis assay. Migrated cells in response to CCL19 were measured by flow cytometry and the percentage of migrated cells is shown. n = 3, line represents mean; *p < 0.05, **p < 0.005, two-way ANOVA with uncorrected fisher’s LSD test. (B) DC obtained from patient PV2 iPS cells as in (A) and from healthy donor iPS cells (healthy control) induce CD4⁺ and CD8⁺ T cell proliferation in MLR assays. CD1c⁺ HLA-DR⁺ cells on day 8–9 of DC differentiation were stimulated with LPS (1 μg/ml, 24 h) and co-cultured with CFSE labeled T cells for 5 days and CD4⁺ and CD8⁺ T cell proliferation was determined by flow cytometry. Representative flow cytometry analysis and quantification of percentage and number of dividing CD4⁺, CD8⁺, and/or CD103⁺ T cells are shown. ConA-stimulated and unstimulated T cells, positive and negative control, respectively. n = 2–3, independent experiments and independent healthy donors; line represents mean; *p < 0.05, **p < 0.005, ***p < 0.001, one-way ANOVA with Tukey’s multiple comparisons test vs. unstimulated T cells or as indicated.