Vaccine driven lung TRM cells provide immunity against Klebsiella via fibroblast IL-17R signaling [scRNA-Seq]

Subunit vaccine elicits lung tissue resident memmory CD4+ T-cells and confers lung protection against heterologous strain Overall design: I. We immunized WT C57BL/6 mice oropharyngealy with 1ug OmpX+10ug LTA1 or 40ug heat-killed Klebsiella pneumoniae (ATCC 43816) twice 3 weeks apart, and 1 more week later we euthanized and removed the lungs for single cell suspensions as well as spleen cells from naïve WT C57BL/6 mice. Then, the CD4+ T-cells were sorted from the cell suspensions by magnetic beads. Recombinant OmpX were generated from K. pneumoniae-43816 and purified by using E. coli expression systems. The adjuvant LTA1 was A1 subunit of the heat-labiled enterotoxin from E.coli and purified from E. coli expression systems. II. We immunized IL-17F-Thy1.1 reporter mice oropharyngealy with 1ug OmpX+10ug LTA1 twice 3 weeks apart, and 1 more week later we euthanized and removed the lungs for single cell suspensions as well as naive spleen cells. Then, the CD4+ T-cells were sorted from the cell suspensions by magnetic beads. Recombinant OmpX were generated from K. pneumoniae-43816 and purified by using E. coli expression systems. The adjuvant LTA1 was A1 subunit of the heat-labiled enterotoxin from E.coli and purified from E. coli expression systems.