Fig. 8.

IFNγ-stimulated lung endothelial cells (LEC) cross-present in vitro PbAluc antigen to reporter cells. a LEC primary culture protocol. Mouse lungs were excised and cut into 1 × 1 × 1 mm³ pieces and placed in a 6-well plate for 60 h. The histograms are representative of two independent culture experiments and show characterization of the endothelial cells by flow cytometry (staining for CD45, CD31, VE-Cadherin (CD144), CD34 and CD62L (L-selectin)). Almost 80% of the cultured cells were CD45⁻CD31⁺ endothelial cells. b LEC seeded in triplicate wells of a 48-well plate were stimulated or not with 10 ng/ml IFN-γ for 24 h and then incubated or not with 5 × 10⁵ thawed PbAluc-matured iRBCs for a further 24 h. The wells were washed and co-cultured with LR-BSL8.4a reporter cells overnight stained for β-gal. The data represent the mean ± SD; **p < 0.01, by ANOVA with Bonferroni’s post-test. ^Statistically significant compared to the first well (negative control) (p < 0.01)