GEO Logo
   NCBI > GEO > Accession DisplayHelp Not logged in | LoginHelp
GEO help: Mouse over screen elements for information.
Series GSE198769 Query DataSets for GSE198769
Status Public on Apr 03, 2022
Title Bronchoalveolar lavage alveolar macrophage subset transcriptomes in WT and NOX2-knockout mice
Organism Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary The leukocyte NADPH oxidase 2 (NOX2) plays a key role in pathogen killing and immunoregulation. Genetic defects in NOX2 result in chronic granulomatous disease (CGD), associated with microbial infections and inflammatory disorders, often involving the lung. Alveolar macrophages (AM) are the predominant immune cell in the airways at steady state, and limiting their activation is important given constant exposure to inhaled materials, yet the importance of NOX2 in this process is not well-understood. Here, we show a previously undescribed role for NOX2 in maintaining lung homeostasis by suppressing AM activation, as studied using CGD mice or mice with selective loss of NOX2 primarily in macrophages. AM lacking NOX2 have increased cytokine responses to TLR2 and TLR4 stimulation ex vivo. Moreover, between 4 and 12 weeks of age, mice with global NOX2 deletion developed an activated CD11bhigh subset of AM with epigenetic and transcriptional profiles reflecting immune activation compared to WT AM. The presence of CD11bhigh AM in CGD mice correlated with increased numbers of alveolar neutrophils and proinflammatory cytokines at steady state as well as increased lung inflammation following insults. Moreover, deletion of NOX2 primarily in macrophages was sufficient for mice to develop an activated CD11bhigh AM subset and accompanying pro-inflammatory sequela. Additionally, we showed that the altered resident macrophage transcriptional profile in the absence of NOX2 is tissue-specific as these changes were not seen in resident peritoneal macrophages. Thus, these data demonstrate that absence of NOX2 in alveolar macrophages leads to their pro-inflammatory remodeling and dysregulates alveolar homeostasis.
Overall design BAL alveolar macrophages were studied from 4 week and 12 week old control and CybbKO mice. For each replicate, BAL AM from various groups of mice were sorted into RPMI+20%FBS on the basis of CD45+Ly6G-SiglecF+CD11c+ markers and CD11b expression (CD11b+or CD11b). To achieve a sufficient number of AM cells (~50,000) per each replicate sample, 2 to 3 mice from same group were pooled. Cells were centrifuged followed by resuspension in RLT-plus RNA Lysis buffer (Qiagen). RNA isolation was done using RNAeasy micro-plus columns (Qiagen). 3 independent replicates from each group were analyzed. RNA seq libraries were generated with Clontech SMARTer kits and sequencing were performed on a HiSeq3000 sequencer.
Contributor(s) Bhattacharya S, Yang W, Magee JA, Dinauer MC
Citation(s) 35357446
Submission date Mar 16, 2022
Last update date Apr 03, 2022
Contact name Mary C Dinauer
Organization name Washington University In St Louis
Department Pathology & Immunology
Street address 660 South Euclid Ave
City St Louis
State/province Missouri
ZIP/Postal code 63110
Country USA
Platforms (1)
GPL21493 Illumina HiSeq 3000 (Mus musculus)
Samples (15)
GSM5956533 cy3l
GSM5956534 cy2l
GSM5956535 cy1l
This SubSeries is part of SuperSeries:
GSE198778 The leukocyte NADPH oxidase 2 (NOX2) plays a key role in pathogen killing and immunoregulation
BioProject PRJNA816851

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE198769_processed_gene_counts.txt.gz 1.4 Mb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

| NLM | NIH | GEO Help | Disclaimer | Accessibility |
NCBI Home NCBI Search NCBI SiteMap