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The role of b2-integrins for the effector functions of polymorphonuclear granulocytes

ABSTRACT: The role of b2-integrins for the effector functions of polymorphonuclear granulocytes

PROVIDER: PRJNA800683 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17761962.fastq.gz	Fastqsanger.gz
	SRR17761963.fastq.gz	Fastqsanger.gz
	SRR17761964.fastq.gz	Fastqsanger.gz
	SRR17761965.fastq.gz	Fastqsanger.gz
	SRR17761966.fastq.gz	Fastqsanger.gz

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The role of b2-integrins for the effector functions of polymorphonuclear granulocytes

Project description:Background: b2-integrins are heterodimeric surface receptors that are expressed specifically by leukocytes and consist of a variable a (CD11a-d) and a common b-subunit (CD18). Functional impairment of CD18, which causes leukocyte adhesion deficiency type-1 results in an immunocompromised state characterized by severe infections, such as invasive pulmonary aspergillosis (IPA). The underlying immune defects have largely been attributed to an impaired migratory and phagocytic activity of polymorphonuclear granulocytes (PMN). However, the exact contribution of b2-integrins for PMN functions in-vivo has not been elucidated yet, since the mouse models available so far display a constitutive CD18 knockout (CD18-/- or CD18hypo). To determine the PMN-specific role of b2-integrins for innate effector functions and pathogen control, we generated a mouse line with a Ly6G-specific knockdown of the common b-subunit (CD18DLy6G). Methods: We analyzed expression levels of genes implicated in b2-integrin signaling for treated and untreated PMN derived from CD18DLy6G mice and CD18fl/fl mice using RNA-Sequencing. To determine the most up- or downregulated genes, genes were sorted on the basis of log2 [fold change] maximum-likelihood estimation, and the P-value cut-off was set to 0.05. Results were illustrated using the pheatmap package. Functional interaction networks were visualized using the STRING package in the open-source platform Cytoscape. Results: Our results revealed a downregulation of b2-integrins , while genes implicated in NFkB singaling were upregulated. With regard to PMN effector function, we observed a downregulation of genes implicated in PMN chemotaxis and microbicidal functions such as Defb40.

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b2 integrins inhibit suppressor of cytokine signaling but also inflammation-associated gene expression in dendritic cells (DC), in line with attenuated T cell activation in autoimmune encephalomyelitis

Project description:Heterodimeric b2 integrin surface receptors (CD11a-d/CD18) are specifically expressed by leukocytes that contribute to pathogen uptake, cell migration, immunological synapse formation and cell signaling. In human, loss of CD18 expression results in leukocyte adhesion deficiency syndrome (LAD-)1, largely characterized by recurrent severe infections.. All available mouse models display a constitutive and ubiquitous knockout of either a or the common b2 (CD18) subunit, which hampers analysis of the cell type-specific role of b2 integrins in vivo. To overcome this limitation, we generated a CD18 gene floxed mouse strain. Offspring generated from crossing with CD11c-Cre mice displayed efficient knockdown of b2 integrins specifically in dendritic cells (DC). Stimulated b2 integrin-deficient splenic DC showed enhanced cytokine production, and concomitantly elevated activity of signal transducers and activators of transcription (STAT) 1, 3 and 5, as well as impaired expression of suppressor of cytokine signaling (SOCS) 2-6 as assessed in bone marrow-derived (BM)DC. Paradoxically,, these BMDC also showed attenuated expression of genes involved in inflammatory signaling. In line, in experimental autoimmune encephalomyelitis mice with a conditional DC-specific b2 integrin knockdown presented with a delayed onset and milder course of disease, associated with lower frequencies of T helper cell (Th)1/Th17 in the inflamed spinal cord. Altogether, our mouse model may prove a valuable tool to study leukocyte-specific functions of b2 integrins in vivo.

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Project description:Polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC) inhibit immune responses in cancer, limit the effects of therapies and promote tumor cell metastasis. Here, we have identified a new precursor that differentiates into granulocytes in vitro and in vivo yet belong to the monocytic lineage. We have termed these cells monocyte-like precursors of granulocytes (MLPG). Under steady state conditions MLPG were absent in the spleen and barely detectable in the bone marrow (BM). In contrast, these cells significantly expanded in tumor-bearing mice. Selective depletion of monocytic cells had no effect on the number of granulocytes present in naïve mice but decreased the population of PMN-MDSC in tumor-bearing mice by 50%. The expansion of MLPG was found to be controlled by the down-regulation of the protein Rb1 and not the IRF8 transcription factor that is known to regulate the expansion of PMN-MDSC from classical granulocytes precursors. In cancer patients, putative MLPG were found within the population of CD15 CD14+HLA-DR-/lo M-MDSC. CXCR1+CD15 CD14+HLA-DR-/lo cells lacked immune suppressive activity but had potential to differentiate to neutrophils in contrast to monocytes and CXCR1- suppressive M-MDSC. These findings describe a mechanism of abnormal myelopoiesis in cancer and suggest potential new approaches for selective targeting of MDSC.

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