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Single-cell RNA sequencing of pulmonary myeloid cells from mixed bone marrow chimeric mice infected with Cryptococcus neoformans gcs1 mutant

ABSTRACT: scRNA-seq of sorted pulmonary myeloid cells (Live/CD45+/IV-/B220-/CD90.2-/Ly-6G-/MHCII+/CD11c+/CD64+) from mixed bone marrow chimeric mice at 35 days post-infection with C. neoformans gcs1 mutant. Chimeric mice were reconstituted with CD45.1 WT : CD45.2 WT (WT chimeras) or CD45.1 WT : CD45.2 STAT6-KO (STAT6-KO chimeras). Cells were labeled with hashtag antibodies (TotalSeq-C, BioLegend) and pooled for library preparation. No sample was excluded due to quality issues.

ORGANISM(S): Mus musculus

PROVIDER: GSE331196 | GEO | 2026/05/18

REPOSITORIES: GEO

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Project description:Innate memory phenotype (IMP) CD4+ T cells are non-conventional Î±Î² T cells exhibiting features of innate immune cells, characterized as CD44high and CD62Llow in periphery. It is recently reported by our group that bone marrow chimeric mice lacking thymic MHCI expression develop predominantly IMP CD8+ T cells, while those lacking hematopoietic MHCI develop predominantly naÃ¯ve CD8+ T cells. Here we perform hirarchical clustering analysis and found that CD4+ T cells share similar property: chimeras lacking thymic MHCII gave rise to predominantly CD4+ T cells that resemble IMP CD4+ T cells observed in WT mice, and vice versa, chimeras lacking hematopoietic MHCII had a majority of naÃ¯ve-like CD4+ T cells resembling naÃ¯veCD4+ T cells seen in WT mice. We used microarrays to compare the global programme of gene expression to determine whether the hematopoietic MHCII selected CD4+ T cells are IMP, and whether the thymic MHCII selected CD4+ T cells are naÃ¯ve CD4+ T cells as observed in WT mice. Through hierarchical clustering and analysis of global gene differential expression, we determined that hematopoietic MHCII dependent IMP CD4+ T cells generated from WT bone marrow transplanted into irradiated MHCII-/- recipients, resemble IMP CD4+ T cells in WT mice, while naÃ¯ve CD4+ T cells generated from MHCII-/- bone marrow transplanted into irradiated WT recipients, resemble naÃ¯ve CD4+ T cells in WT mice. Cell Sorting was performed using a Cytopeia Influx Cell Sorter. Chimeric IMP (CD45.1+TCRÎ²+CD4+CD44highCD62Llow) CD4+ T cells were sorted from splenocytes of CD45.1+WTâCD45.2+MHCII-/- chimeras (WM IMP CD4), and chimeric naÃ¯ve (CD45.2+TCRÎ²+CD4+CD44lowCD62Lhigh) CD4+ T cells were sorted from splenocytes of CD45.2+MHCII-/- â CD45.1+WT chimeras (MW naÃ¯ve CD4) respectively, 8 weeks post transplantation. WT IMP (TCRÎ²+CD4+CD44highCD62Llow) and naÃ¯ve (TCRÎ²+CD4+CD44lowCD62Lhigh) CD4+ T cells were sorted from splenocytes of 8-week old WT mice.

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Single-cell RNA sequencing of pulmonary myeloid cells from mixed bone marrow chimeric mice infected with Cryptococcus neoformans gcs1 mutant

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