Project description:Natural killer (NK) cells function by eliminating virus-infected cells or tumor cells. Here, we identified a NK lineage-biased progenitor population, termed early NK progenitors (ENKP), which developed into NK cells independently of common precursors for ILCs (ILCPs). ENKP-derived NK cells (ENKP_ NK cells) and ILCP-derived NK cells (ILCP_ NK cells) were also transcriptionally different. We devised combinations of surface markers that identified highly enriched ENKP_NK and ILCP_NK cell populations in wild-type mice. Furthermore, Ly49H+ NK cells that responded to mouse cytomegalovirus (MCMV) infection primarily developed from ENKPs whereas ILCP_NK cells were better IFN-g producers upon Salmonella and Herpes Simplex Virus (HSV) infections. Interestingly, human CD56dim and CD56bright NK cells were transcriptionally similar to ENKP_NK cells and ILCP_NK cells, respectively. Our findings establish the existence of two pathways of NK cell development that generate functionally distinct NK cell subsets in mice, and further suggest these pathways may be conserved in humans.
Project description:Investigation of global gene expression levels between B cells, Natural killer cells and Natural killer B cells Gene expression profiling using sorted B cells, Natural killer cells and Natural killer B cells from WT mouse spleen. Total RNA extracted from WT cells were quantified by the NanoDrop ND-1000 and RNA integrity was assessed by standard denaturing agarose gel electrophoresis. The sample preparation and microarray hybridization were performed based on the NimbleGenâs standard protocols.
Project description:Comparing global gene expression of neonatal and adult natural killer cells to determine if differences in gene expression suggest that different developmental pathways during hematopoiesis are followed in the fetal and adult mouse to produce mature natural killer cells.
Project description:The placenta an essential extra-embryonic organ that supports the fetus throughout gestation. The interactions between the placenta and the maternal immune system during the first trimester have not been completely characterized, despite their close physical association and hem-allogeneic relationship to each other. The most abundant type of immune cell found in the uterus in the first trimester is the decidual natural killer cell (dNK). Despite their name, dNKs play supportive roles during pregnancy by remodelling uterine spiral arteries. We present evidence suggesting that the matrix metalloproteinases (MMPs) that dNKs secrete to promote this remodelling also drive placental development. This study was performed using a novel co-culture system of dNKs and trophoblast organoids, which are mini-organs that represent two to three different cell types of the human placenta. We found that co-cultures for one week led to significant (p=0.020) increases in organoid area. Through bulk RNA sequencing and immunohistochemical examinations we also observed significant decreases in trophoblast stemness markers, and upregulation of gene sets associated with extravillous trophoblast (EVT) development. These changes were accompanied by significant (p<0.001) increases in collagen subunit gene expression in the organoids, with simultaneous significant decreases (p<0.001) in the proportion of organoid area occupied by collagen as determined through Masson’s Trichrome. Cultures containing dNKs also contained significantly higher levels of MMP1, 3, 9, and 10 in their culture media, each of which can break down collagen. Collectively, these findings demonstrate that dNKs promote changes concordant with trophoblast differentiation towards EVTs and villous branching morphogenesis.
Project description:Human peripheral blood natural killer cells are grouped in various ways according to phenotypic and functional characteristics. We sorted peripheral blood natural killer cells from healthy donors into seven surface phenotype-defined populations, hashtagged them, and sequenced them. This analysis permitted simultaneous identification of transcriptional clusters and traceback of the components to their phenotypic groups.