IL-36 signaling in keratinocytes controls early IL-23 production in psoriasis-like dermatitis

Jérémie D Goldstein; Esen Y Bassoy; Assunta Caruso; Jennifer Palomo; Emiliana Rodriguez; Sylvain Lemeille; Cem Gabay

doi:10.26508/lsa.202000688

IL-36 signaling in keratinocytes controls early IL-23 production in psoriasis-like dermatitis

Life Science Alliance ◽

10.26508/lsa.202000688 ◽

2020 ◽

Vol 3 (6) ◽

pp. e202000688 ◽

Cited By ~ 7

Author(s):

Jérémie D Goldstein ◽

Esen Y Bassoy ◽

Assunta Caruso ◽

Jennifer Palomo ◽

Emiliana Rodriguez ◽

...

Keyword(s):

Crucial Role ◽

Early Time ◽

Neutrophil Infiltration ◽

Specific Function ◽

Facs Analysis ◽

Time Points ◽

Qrt Pcr ◽

Early Production

IL-36R signaling plays an important role in the pathogenesis of psoriasis. We ought to assess the specific function of IL-36R in keratinocytes for the pathology of Aldara-induced psoriasis-like dermatitis. Il36rΔK mice presenting deletion of IL-36R in keratinocytes were similarly resistant to Aldara-induced ear inflammation as Il36r−/− mice, but acanthosis was only prevented in Il36r−/− mice. FACS analysis revealed that IL-36R signaling in keratinocytes is mandatory for early neutrophil infiltration in Aldara-treated ears. RNASeq and qRT-PCR experiments demonstrated the crucial role of IL-36R signaling in keratinocytes for induction of IL-23, IL-17, and IL-22 at early time points. Taken together, our results demonstrate that IL-36R signaling in keratinocytes plays a major role in the induction of Aldara-induced psoriasis-like dermatitis by triggering early production of IL-23/IL-17/IL-22 cytokines and neutrophil infiltration.

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The role of microRNA-3085 in chondrocyte function

Scientific Reports ◽

10.1038/s41598-020-78606-6 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Linh Le ◽

Lingzi Niu ◽

Matthew J. Barter ◽

David A. Young ◽

Tamas Dalmay ◽

...

Keyword(s):

Feedback Inhibition ◽

Interleukin 1 ◽

Early Time ◽

Null Cell ◽

Time Points ◽

Vitro Analysis ◽

In Vitro Analysis ◽

Cultured Chondrocytes

AbstractMicroRNAs have been shown to play a role in cartilage development, homeostasis and breakdown during osteoarthritis. We previously identified miR-3085 in humans as a chondrocyte-selective microRNA, however it could not be detected by Northern blot. The aim of the current study was to prove that miR-3085 is a microRNA and to investigate the function of miR-3085 in signaling pathways relevant to cartilage homeostasis and osteoarthritis. Here, we confirm that miR-3085 is a microRNA and not another class of small RNA using (1) a pre-miR hairpin maturation assay, (2) expression levels in a Dicer null cell line, and (3) Ago2 pulldown. MicroRNA-3085-3p is expressed more highly in micromass than monolayer cultured chondrocytes. Transfection of miR-3085-3p into chondrocytes decreases expression of COL2A1 and ACAN, both of which are validated as direct targets of miR-3085-3p. Interleukin-1 induces the expression of miR-3085-3p, at least in part via NFκB. In a feed-forward mechanism, miR-3085-3p then potentiates NFκB signaling. However, at early time points after transfection, its action appears to be inhibitory. MyD88 has been shown to be a direct target of miR-3085-3p and may be responsible for the early inhibition of NFκB signaling. However, at later time points, MyD88 knockdown remains inhibitory and so other functions of miR-3085-3p are clearly dominant. TGFβ1 also induces the expression of miR-3085-3p, but in this instance, it exerts a feedback inhibition on signaling with SMAD3 and SMAD4 shown to be direct targets. This in vitro analysis shows that miR-3085-3p functions in chondrocytes to induce IL-1-signaling, reduce TGFβ1 signaling, and inhibit expression of matrix genes. These data suggest that miR-3085-3p has a role in chondrocyte function and could contribute to the process of osteoarthritis.

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Role of C1q in spleen-derived neutrophil infiltration and neuroinflammation after ICH in mice

10.21203/rs.2.18353/v1 ◽

2019 ◽

Author(s):

Jun Tang ◽

Tiantian Luo ◽

Junjun Geng ◽

Bo Zhang ◽

Yin Niu ◽

...

Keyword(s):

Neutralizing Antibody ◽

Neutrophil Infiltration ◽

Inflammatory Factors ◽

Secondary Brain Injury ◽

Neuron Death ◽

Qrt Pcr ◽

Propidium Iodide Staining ◽

Tnf Α ◽

Targeted Inhibition

Abstract Background: Neuroinflammation is a major detrimental role of secondary brain injury after spontaneously intracerebral hemorrhage (ICH). Neutrophil infiltration plays a key role in the pathophysiology of ICH, but the coming resource and mechanism is unknown. This study aims to investigate whether spleen-derived neutrophil infiltration accelerated neuroinflammation and the role of C1q classical pathway.Methods: Male C57 mice were subjected to collagenase-induced ICH. If necessary, splenectomy was performed 2 weeks prior to ICH induction or anti-C1q neutralizing antibody (50mg/kg) was injected intravenously into the tail vein 15 minutes prior. Immunohistochemistry, Propidium Iodide staining, western blotting, ELISA and qRT-PCR were used to study the change of molecular proteins, neuronal cells and inflammatory factors. 7.0T animal MRI was used to assess hydrocephalus.Results: At 0h, 6h, 12h, 24h and 48h post ICH induction, we found a significant increasing tendency of microglia activation and neutrophil infiltration around hematoma and that C1q upregulation was correlated with neuronal decrease, which peaked at 24h after ICH. Here, we demonstrated spleen atrophy and upregulation of neutrophil in spleen 24h after ICH. Splenectomy prior to ICH mice resulted in significant decrease of microglia and neutrophil infiltration compared with that in group of sham-splenectomy. Moreover, both anti-C1q antibody and splenectomy significantly attenuated neutrophil infiltration and neuron death, restored synapse VGAT, alleviated hydrocephalus and inflammatory factors, such as IL-1β, TNF-α and IL-6 after ICH.Conclusion: The study demonstrated that spleen is a major source of brain neutrophil infiltration after ICH. C1q-targeted inhibition of classic complement pathway could prevent spleen-derived neutrophil infiltration and attenuate ICH induced neuroinflammation, which provides a novel therapeutic approach for hemorrhagical stroke.

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Effects of ATP and bradykinin on endothelial cell Ca2+ homeostasis and formation of cGMP and prostacyclin

AJP Cell Physiology ◽

10.1152/ajpcell.1993.265.6.c1620 ◽

1993 ◽

Vol 265 (6) ◽

pp. C1620-C1629 ◽

Cited By ~ 27

Author(s):

E. C. Gosink ◽

E. J. Forsberg

Keyword(s):

Nitric Oxide ◽

Early Time ◽

Receptor Occupancy ◽

Cell Types ◽

No Synthase ◽

Time Points ◽

No Formation ◽

Per Se ◽

Cgmp Formation

ATP and bradykinin are known to activate Ca2+ release from intracellular Ca2+ pools as well as induce the influx of Ca2+ in many cell types. In adrenal medulla endothelial cells, we found that ATP and bradykinin could activate Ca2+ influx, although Ca2+ influx did not appear to be due to depletion of intracellular Ca2+ pools per se, since depletion of intracellular Ca2+ pools with thapsigargin reduced rather than enhanced both unidirectional and steady-state 45Ca2+ uptake. In addition, Ca2+ influx, activated by ATP but not bradykinin, was mostly abolished after agonist removal in cells in which intracellular Ca2+ pools had not been allowed to refill, suggesting that continued receptor occupancy was necessary for ATP to activate Ca2+ influx. The role of Ca2+ in activating guanosine 3',5'-cyclic monophosphate (cGMP) formation [a marker for nitric oxide (NO) secretion] and prostacyclin (PGI2) secretion was also studied. Bradykinin-induced cGMP and PGI2 formation and ATP-induced PGI2 formation each required Ca2+ release from intracellular Ca2+ pools, since depletion of these pools with thapsigargin inhibited their formation. In contrast, ATP-induced cGMP formation, particularly at early time points, did not appear to require either Ca2+ release or Ca2+ influx. This suggests that ATP, but not bradykinin, either induces Ca(2+)-independent NO formation or that ATP stimulates the generation of cGMP independently of NO. The latter supposition is supported by our observation that NO synthase inhibitors inhibited ATP-induced cGMP formation by at most 50%.

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Listeria monocytogenes Flagella Are Used for Motility, Not as Adhesins, To Increase Host Cell Invasion

Infection and Immunity ◽

10.1128/iai.00886-06 ◽

2006 ◽

Vol 74 (12) ◽

pp. 6675-6681 ◽

Cited By ~ 87

Author(s):

Heather S. O'Neil ◽

Hélène Marquis

Keyword(s):

Listeria Monocytogenes ◽

Cell Invasion ◽

Early Time ◽

Host Cells ◽

Host Cell Invasion ◽

Time Points ◽

Food Borne ◽

Epithelial Cell Invasion

ABSTRACT Flagellar structures contribute to the virulence of multiple gastrointestinal pathogens either as the effectors of motility, as adhesins, or as a secretion apparatus for virulence factors. Listeria monocytogenes is a food-borne, gram-positive pathogen that uses flagella to increase the efficiency of epithelial cell invasion (A. Bigot, H. Pagniez, E. Botton, C. Frehel, I. Dubail, C. Jacquet, A. Charbit, and C. Raynaud, Infect. Immun. 73:5530-5539, 2005; L. Dons, E. Eriksson, Y. Jin, M. E. Rottenberg, K. Kristensson, C. N. Larsen, J. Bresciani, and J. E. Olsen, Infect. Immun. 72:3237-3244, 2004). In this study, we aimed to elucidate the mechanism by which flagella contribute to L. monocytogenes invasion. To examine the role of flagella as adhesins, invasion and adhesion assays were performed with flagellated motile and nonmotile bacteria and nonflagellated bacteria. We observed that flagellated but nonmotile bacteria do not adhere to or invade human epithelial cells more efficiently than nonflagellated bacteria. These results indicated that flagella do not function as adhesins to enhance the adhesion of L. monocytogenes to targeted host cells. Instead, it appears that motility is important for tissue culture invasion. Furthermore, we tested whether motility contributes to early colonization of the gastrointestinal tract using a competitive index assay in which mice were infected orally with motile and nonmotile bacteria in a 1:1 ratio. Differential bacterial counts demonstrated that motile bacteria outcompete nonmotile bacteria in the colonization of the intestines at early time points postinfection. This difference is also reflected in invasion of the liver 12 h later, suggesting that flagellum-mediated motility enhances L. monocytogenes infectivity soon after bacterial ingestion in vivo.

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Transcriptome-Based Analysis Reveals a Crucial Role of BxGPCR17454 in Low Temperature Response of Pine Wood Nematode (Bursaphelenchus xylophilus)

International Journal of Molecular Sciences ◽

10.3390/ijms20122898 ◽

2019 ◽

Vol 20 (12) ◽

pp. 2898 ◽

Cited By ~ 3

Author(s):

Bowen Wang ◽

Xin Hao ◽

Jiayao Xu ◽

Yan Ma ◽

Ling Ma

Keyword(s):

Low Temperature ◽

Rna Sequencing ◽

Crucial Role ◽

Temperature Response ◽

Bursaphelenchus Xylophilus ◽

Sequencing Analysis ◽

Pine Wood Nematode ◽

Pine Wood ◽

Qrt Pcr

Background: The causal agent of pine wilt disease is the pine wood nematode (PWN) (Bursaphelenchus xylophilus), whose ability to adapt different ecological niches is a crucial determinant of their invasion to colder regions. To discover the molecular mechanism of low temperature response mechanism, we attempted to study the molecular response patterns under low temperature from B. xylophilus with a comprehensive RNA sequencing analysis and validated the differentially expressed genes (DEGs) with quantitative real-time polymerase chain reaction (qRT-PCR). Bioinformatic software was utilized to isolate and identify the low-temperature-related BxGPCR genes. Transcript abundance of six low-temperature-related BxGPCR genes and function of one of the BxGPCR genes are studied by qRT-PCR and RNA interference. Results: The results showed that we detected 432 DEGs through RNA sequencing between low-temperature-treated and ambient-temperature-treated groups nematodes. The transcript level of 6 low-temperature-related BxGPCR genes increased at low temperature. And, the survival rates of BxGPCR17454 silenced B. xylophilus revealed a significant decrease at low temperature. Conclusion: in conclusion, this transcriptome-based study revealed a crucial role of BxGPCR17454 in low temperature response process of pine wood nematode. These discoveries would assist the development of management and methods for efficient control of this devastating pine tree pest.

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Differential roles for β2 integrins in experimental autoimmune bullous pemphigoid

Blood ◽

10.1182/blood-2005-08-3123 ◽

2006 ◽

Vol 107 (3) ◽

pp. 1063-1069 ◽

Cited By ~ 27

Author(s):

Zhi Liu ◽

Minglang Zhao ◽

Ning Li ◽

Luis A. Diaz ◽

Tanya N. Mayadas

Keyword(s):

Bullous Pemphigoid ◽

Early Time ◽

Neutralizing Antibody ◽

Neutrophil Infiltration ◽

Neutrophil Accumulation ◽

Neutrophil Degranulation ◽

Apoptosis Markers ◽

Β2 Integrins ◽

Experimental Autoimmune

AbstractBullous pemphigoid (BP) is an autoimmune disease associated with autoantibodies directed against the hemidesmosomal antigens anti-BP230 and anti-B180. Neonatal mice injected with rabbit anti-mouse BP180 (mBP10) IgG develop a BP-like disease. Complement, immune complexes, mast cells, and neutrophils play a key role in subepidermal blistering in this animal model. In this study we investigated the role of β2 integrins in experimental BP. Wild-type (WT) mice pretreated with neutralizing antibody against CD11a (LFA-1), CD11b (Mac-1), CD11a plus CD11b, or CD18 alone failed to develop BP when injected with pathogenic anti-mBP180 IgG. This was associated with a significant reduction in neutrophil accumulation in neutralizing antibody-treated mice. Mac-1-deficient (Mac-1 knockout [KO]) mice were resistant to experimental BP despite normal complement deposition and mast cell and neutrophil degranulation. Neutrophil infiltration in Mac-1 KO mice was severely impaired at 24 hours. However, more neutrophils accumulated in the skin of Mac-1 KO mice compared with WT mice at early time points (2-4 hours), which was associated with an increase in their survival as determined by apoptosis markers. These data suggest that β2 integrins play differential roles in experimental BP: LFA-1 is required for neutrophil recruitment, while Mac-1 mediates late neutrophil accumulation and apoptosis of infiltrating neutrophils.

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