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. Author manuscript; available in PMC: 2012 Dec 23.
Published in final edited form as: Immunity. 2011 Dec 23;35(6):997–1009. doi: 10.1016/j.immuni.2011.10.018

Figure 5.

Figure 5

Cross-serotype protection in heterologous challenge is mediated by Th17 not Th1. (A) C57BL/6 and Ifng−/− mice were immunized intranasally with 20 µg heat-killed K. Pneumoniae-43816 (ATCC) (serotype K2) at Day 0 and Day 7. 28 days after the 2nd immunization, immunized mice and naïve-unimmunized mice were infected with 104 K. pneumoanie-396 (serotype K1) and sacrificed 24h after. Lung burden and systemic dissemination were determined by lung (left panel) and spleen (right panel) CFUs. Each group has 4–5 mice. (B) IL-17F-Thy1.1 reporter mice were immunized with heat-killed K. Pneumoniae-43816 (ATCC) (serotype K2) and were sacrificed one week after. Lung single cells were prepared and stained with CD4 and Thy1.1. flow cytometry sorted 105 Th17 (CD4+Thy1.1+) per mouse or 105 Th-non17F (CD4+Thy1.1−) per mouse were intravenously transferred to Rag2−/−Il2rg−/− mice. Four week after transfer, mice were infected with 104 K. Pneumoniae-396 (serotype K1) and sacrificed at 24h. Lung burdens were assessed by CFU. Each group has 3–4 mice. (C) IL-17F-Thy1.1 reporter mice were immunized with heat-killed K. Pneumoniae-43816 (ATCC) (serotype K2) and were sacrificed one week after. Lung single cells were prepared and stained with CD4 and Thy1.1. flow cytometry sorted 105 Th17 (CD4+Thy1.1+) per mouse were intravenously transferred to Rag2−/− mice. Four week after transfer, mice were infected with 104 K. Pneumoniae-396 (serotype K1). Control IgG, α-IL17RC or α-IFN-γ were given i.t. 12h after infection and mice were sacrificed at 24h after infection. Lung and spleen burdens were assessed by CFU. Each group has 4 mice. (D) Il17ra−/− mice with B-cell depletion (0.1mg anti-CD20 i.p. weekly) mice were immunized intranasally with 20µg heat-killed K. Pneumoniae-43816 (ATCC) (serotype K2) at Day 0 and challenge with 104 K. Pneumoniae-396 (serotype K1) on Day28. Control IgG or α-IFN-γ were given i.t. 12h after infection and mice were sacrificed at 24h after infection. Lung bacterial burdens were assessed by CFU. Each group has 4–5 mice.