Figure 3.

ILCs and ILC2P cells emerge to the site of inflammation independently of HSPCs in response to high-dose of zymosan treatment. WT or CCR2 KO mice were injected with 0,1 mg (low dose) or 10 mg (high dose) of zymosan and 24 h later peritoneal cells were harvested, stained for markers identifying both mature ILCs and progenitor cells of type 2 ILCs. (A) CCR2 KO mice where bone marrow-derived HSPCs are unable to migrate to the site of sterile inflammation, were injected either with a low- or high-dose of zymosan. Twenty-four hours post-injection, cells were recovered from the peritoneum of injected mice and stained for Lineage markers, CD45 and CD127. One representative experiment of flow cytometry analysis is shown from 4 independent experiments (n = 4) (B) Statistical analysis of the frequency of Lineage⁻CD127⁺ cells recovered from the peritoneal cavity of zymosan-injected mice (CCR2 KO recipients). The same experimental process as described in (A) was used to investigate the emergence of bone marrow-derived progenitor cells for type 2 innate lymphoid cells in WT mice treated either with a low- or high-dose of zymosan. (C) Cells were stained for CD45, Lineage markers, CD127, CD25, Sca-1, and KLRG-1 (n = 4). (D) Intraperitoneal cells were recovered from the peritoneal cavity of both high- and low-dose treated CCR2 KO mice and stained for markers of ILC2 and ILC2P as described in (B). One representative experiment is shown (n = 4). (E) Statistical analysis of the frequency of CD25⁺Sca-1⁺ cells gated previously in CD45⁺Lineage⁻CD127⁺ cells of both WT and CCR2 KO mice injected either with PBS (control) or low- and high-dose of zymosan (n = 4). (F) Statistical analysis of the frequency of Sca-1⁺KLRG1⁺ and Sca-1⁺KLRG1⁻ cells from mice injected either with PBS or low- and high-dose zymosan both in WT and CCR2 KO mice (n = 4). *p < 0.05.