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RNA-seq of spleen monocytes from IL-4Rα+/+ and IL-4Rα-/- mice on a C57BL/6J background

ABSTRACT: Monocytes are immune cells which differentiate into subsets through sequential transition. The life-time differs between the individual subsets ranging from 1 day to 1 week. We studied the involvement of IL-4 receptor signaling on monocyte survival in mice lacking IL-4 receptors in a total knockout and a myeloid knockout. Circulating leukocytes were reduced by 25% in both knockouts, which was mainly caused by a 50% reduction of monocytes. Using RNA sequencing we were able to demonstrate that spleen monocytes in IL-4Rα deficient mice express lower amounts of anti-apoptotic genes and increased amounts of cell adhesion molecules hinting to an increase in apoptotic potential of these cells. This data were also verified in spleen samples were we found an increase of apoptotic monocytes identified by TUNEL and cleaved caspase 3 staining. Mechanistically we were able to show that IL-4 signaling directly affects the lifespan of monocytes in vivo. Therefore, we identified a previously unknown function of basal IL-4 levels in homeostasis of monocytes by regulating the lifespan and preventing apoptosis of the cells.

ORGANISM(S): Mus musculus

PROVIDER: GSE168143 | GEO | 2022/09/01

REPOSITORIES: GEO

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Project description:Selective stimulation of IL-4 receptor on smooth muscle induces airway hyper-responsiveness in mice. Abstract: Production of the cytokines IL-4 and IL-13 is increased in both human asthma and mouse asthma models and Stat6 activation by the common IL-4/IL-13R drives most mouse model pathophysiology, including airway hyperresponsiveness (AHR). However, the precise cellular mechanisms through which IL-4Rα induces AHR remain unclear. Overzealous bronchial smooth muscle constriction is thought to underlie AHR in human asthma, but the smooth muscle contribution to AHR has never been directly assessed. Furthermore, differences in mouse vs. human airway anatomy and observations that selective IL-13 stimulation of Stat6 in airway epithelium induces murine AHR raise questions about the importance of direct IL-4R effects on smooth muscle in murine asthma models and relevance of these models to human asthma. Using transgenic mice in which smooth muscle is the only cell type that expresses or fails to express IL-4Rα, we demonstrate that direct smooth muscle activation by IL-4, IL-13, or allergen is sufficient, but not necessary, to induce AHR and show that 5 genes known to promote smooth muscle migration, proliferation and contractility are activated by IL-13 in smooth muscle in vivo. These observations demonstrate that IL-4Rα promotes AHR through multiple mechanisms and provide a model for testing smooth muscle-directed asthma therapeutics. For the microarray aspect of of the study, there were three groups of mice: 1. IL4R gene knockout (KO) mice 2. WT mice 3. IL4R KO mice that were also transgenic for a gene construct that expressed IL4R under the control of the smooth muscle-specific promoter from the SMP8 gene All mice were subjected to intratracheal IL13 exposure for 7 days, and whole lung RNA was prepared for microarray analysis 24 hours after the last instillation. Per treatment and genotype: Two RNA pools were made from four mice each. These were labeled and hybridized to make a total of 6 microarrays. RNA was labeled with the standard Affymetrix 3' labeling protocol to make cDNA that was hybridized to Mouse MOE 430 plus 2.0 GeneChips. Gene transcripts were identified that differed in their relative expression as a function of IL4R expression on the smooth muscle cells.

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RNA-seq of spleen monocytes from IL-4Rα+/+ and IL-4Rα-/- mice on a C57BL/6J background

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