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ADAP-STAT1 module maintains platelet homeostasis via restraint of macrophage phagocytic in ITP

ABSTRACT: ADAP-STAT1 module maintains platelet homeostasis via restraint of macrophage phagocytic in ITP

PROVIDER: PRJNA760470 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15712730_1.fastq.gz	Fastqsanger.gz
	SRR15712730_2.fastq.gz	Fastqsanger.gz
	SRR15712731_1.fastq.gz	Fastqsanger.gz
	SRR15712731_2.fastq.gz	Fastqsanger.gz
	SRR15712732_1.fastq.gz	Fastqsanger.gz

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ADAP-STAT1 module maintains platelet homeostasis via restraint of macrophage phagocytic in ITP

Project description:Heightened platelet phagocytosis by macrophages accompanied with an increase in IFN-γ is often attributed to the etiology of immune thrombocytopenia (ITP), an autoimmune disease characterized by low platelet counts. While deletion of the hematopoietic cell adaptor protein ADAP predisposes to ITP in mice, the mechanisms that underlie the functional role of ADAP during ITP remains largely unknown. Here we report that ADAP restrains platelet clearance by macrophages via modulation of STAT1-FcγR signaling axis. We show that under-expression of ADAP in splenic macrophage was associated with low platelet counts in patients with ITP. Further, macrophages from Adap−/− mice had an elevated phagocytosis capacity and exhibited upregulated pro-inflammatory signaling, and the thrombocytopenia in Adap−/− mice was mitigated in vivo by depletion of macrophages. Mechanically, ADAP interacted and competed with STAT1 binding to importin a5, a nuclear shuttling receptor for activated STAT1. While having no effect on STAT1 tyrosine phosphorylation, loss of ADAP increased the assembly of STAT1-importin a5 complex and facilitated STAT1 nuclear entry, leading to a selective enhancement of transcription of FcγRI/IV in macrophages. Moreover, pharmacological inhibition of STAT1 or disruption of STAT1-importin a5 interaction relieved the thrombocytopenia in Adap−/− mice. Thus, our findings reveal a critical role for ADAP as an intracellular immune checkpoint for shaping macrophage phagocytic ability in homeostatic maintenance of platelets.

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Acute ITP vs. Chronic ITP (Children)

Project description:Whole blood transcriptional profiling of pediatric idiopathic thrombocytopenic purpura (ITP) patients comparing self-limited acute ITP patients with chronic ITP patients or progression-to-chronic ITP patients. Patient samples were collected during 3 different disease states: 8 samples from acute ITP pts (within 6 months of dx, during active disease), 4 samples from progression-to-chronic ITP pts (within 6 months of dx), and 14 samples from chronic ITP (after 6 months of dx).

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Project description:CD3+ cells were studied in duplicate from healthy controls, patients with active ITP and ITP in remission. Keywords: repeat sample

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Project description:Five ITP and five control whole blood samples were collected with IRB approval. The age range of ITP patients was 1-54 years, and the range in platelet counts was 2,000-98,000 W 109/L. We examined only one ITP sample for antiplatelet antibodies, and found it to be positive. Total RNA was isolated from 2.5 ml whole blood according to the PAXgene Blood RNA System (PreAnalytix, Switzerland). RNA isolated from ITP patients and controls, along with Human Universal Reference RNA (Stratagene, La Jolla, CA) were linearly amplified using the MessageAmp aRNA amplification kit (Ambion, Austin, TX). Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Keywords: Logical Set

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Project description:The ability of Entamoeba histolytica to phagocytose host cells correlates to observed virulence in vivo. To better understand the mechanism of phagocytosis we used paramagnetic beads coated with host ligand and sorted trophozoites based on phagocytic ability. Gene expression was then measured in both the sorted phagocytic and non-phagocytic populations using a custom Affymetrix chip for E. histolytica. Feed forward regulation of phagocytosis by Entamoeba histolytica. Infection and Immunity. PMID 23045476 M280 Streptavidin Dynabeads (Invitrogen) were labeled with 20ug/mL biotinylated Human C1q (Quidel). Entamoeba histolytica (strain HM1) were washed twice with PBS then resuspended with the labeled beads at a 10:1 ratio of beads to trophozoites. Samples were incubated at 37Â°C for 45 minutes, washed twice with agitation to remove adherent beads, then resuspended in MACS buffer. Samples were loaded into magnetic columns (Miltenyi Biotec) and trophozoites were seperated according to manufacturer's protocols. phagocytic vs. non-phagocytic Entamoeba histolytica populations

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Project description:CD3+ cells were studied in duplicate from healthy controls, patients with active ITP and ITP in remission.

2007-09-28 | E-GEOD-574 | biostudies-arrayexpress