Wnt5a–Rac1–NF-κB Homeostatic Circuitry Sustains Innate Immune Functions in Macrophages

Naskar, Debdut; Maiti, George; Chakraborty, Arijit; Roy, Arunava; Chattopadhyay, Dhrubajyoti; Sen, Malini

doi:10.4049/jimmunol.1302817

Cited by 70 publications

(126 citation statements)

References 68 publications

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“…Several lines of evidence suggest that a basal level of Wnt5a signaling contributes to the sustainment of host immune responses (16). Because L. donovani infection disrupts immune homeostasis, it was important to determine whether the steady-state level of Wnt5a expression is altered in L. donovani-infected macrophages.…”

Section: Donovani Infection Leads To Disruption Of the Steady-statmentioning

confidence: 99%

“…The effect of Wnt5a was blocked by the addition of Rac1i or ROCK1i 3 h after the addition of Wnt5a (15,16). To assess the effect of IWP-2 on infection, plated RAW 264.7 cells or BMDMs were treated with 0.05 mM IWP-2 or DMSO for 48 h prior to infection for 4 h. To determine the effect of Wnt5a postinfection, RAW 264.7 cells infected for 4 h were treated with 50 ng/ml Wnt5a for 6 h. To determine the effect of L. donovani infection on the steady-state level of Wnt5a, similarly plated RAW 264.7 macrophages were infected for 4 h with L. donovani promastigotes/axenic amastigotes at an MOI ∼ Subsequently, cells were treated with 0.5 mM O-phenanthroline (inhibitor of L. donovani GP63-like protease) for 2 h to for check the inhibition of L. donovani-specific metalloprotease activity.…”

Section: Estimation Of Leishmania Infection In Macrophagesmentioning

confidence: 99%

“…siRNA (25 nM) was complexed with 2.5 ml of Lipofectamine RNAiMAX transfection reagent (Life Technologies) in 200 ml of serumfree antibiotic-free RPMI 1640 in a sterile microcentrifuge tube for 30 min, after which 0.8 ml of RPMI 1640 medium containing 10% FBS was added. The cells in each well were incubated with 1 ml of transfection mix for 24 h. The detailed procedure was described previously (15,16).…”

Section: Small Interfering Rna Transfectionmentioning

confidence: 99%

“…Bone marrow-derived macrophages (BMDMs) and peritoneal macrophages (PMFs) were prepared for assays following a previously published protocol (16).…”

Section: Reagentsmentioning

confidence: 99%

“…Because Wnts, particularly Wnt5a and Wnt3a, influence cytoskeletal dynamics (12)(13)(14), it was important to investigate whether L. donovani PV formation or its sustainment is influenced by Wnt signaling. In keeping with the demonstrated role of Wnt5a signaling in the sustainment of immune homeostasis (15,16), we studied the influence of Wnt5a signaling on L. donovani infection.…”

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confidence: 99%

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Wnt5a Signaling Promotes Host Defense againstLeishmania donovaniInfection

Chakraborty

Kurati

Mahata

et al. 2017

The Journal of Immunology

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Leishmania donovani infects macrophages, disrupting immune homeostasis. The underlying mechanism that sustains infection remains unresolved. In view of the potential of Wnt5a signaling to support immune homeostasis, we evaluated the interrelationship of Wnt5a signaling and Leishmania donovani infection. Upon infecting macrophages separately with antimony drug–sensitive and –resistant L. donovani, we noted disruption in the steady-state level of Wnt5a. Moreover, inhibition of Wnt5a signaling by small interfering RNA transfection in vitro or by use of inhibitor of Wnt production in vivo led to an increase in cellular parasite load. In contrast, treatment of macrophages with recombinant Wnt5a caused a decrease in the load of antimony-sensitive and -resistant parasites, thus confirming that Wnt5a signaling antagonizes L. donovani infection. Using inhibitors of the Wnt5a signaling intermediates Rac1 and Rho kinase, we demonstrated that Wnt5a-mediated inhibition of parasite infection in macrophages is Rac1/Rho dependent. Furthermore, phalloidin staining and reactive oxygen species estimation of Wnt5a-treated macrophages suggested that a Wnt5a-Rac/Rho–mediated decrease in parasite load is associated with an increase in F- actin assembly and NADPH oxidase activity. Moreover, live microscopy of L. donovani–infected macrophages treated with Wnt5a demonstrated increased endosomal/lysosomal fusions with parasite-containing vacuoles (parasitophorous vacuoles [PV]). An increase in PV–endosomal/lysosomal fusion accompanied by augmented PV degradation in Wnt5a-treated macrophages was also apparent from transmission electron microscopy of infected cells. Our results suggest that, although L. donovani evades host immune response, at least in part through inhibition of Wnt5a signaling, revamping Wnt5a signaling can inhibit L. donovani infection, irrespective of drug sensitivity or resistance.

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Section: Donovani Infection Leads To Disruption Of the Steady-statmentioning

confidence: 99%

Section: Estimation Of Leishmania Infection In Macrophagesmentioning

confidence: 99%

Section: Small Interfering Rna Transfectionmentioning

confidence: 99%

“…Bone marrow-derived macrophages (BMDMs) and peritoneal macrophages (PMFs) were prepared for assays following a previously published protocol (16).…”

Section: Reagentsmentioning

confidence: 99%

mentioning

confidence: 99%

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Wnt5a Signaling Promotes Host Defense againstLeishmania donovaniInfection

Chakraborty

Kurati

Mahata

et al. 2017

The Journal of Immunology

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WNT-5A: signaling and functions in health and disease

Kumawat

Gosens

2015

Cell. Mol. Life Sci.

135

115

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WNT-5A plays critical roles in a myriad of processes from embryonic morphogenesis to the maintenance of post-natal homeostasis. WNT-5A knock-out mice fail to survive and present extensive structural malformations. WNT-5A predominantly activates β-catenin-independent WNT signaling cascade but can also activate β-catenin signaling to relay its diverse cellular effects such as cell polarity, migration, proliferation, cell survival, and immunomodulation. Moreover, aberrant WNT-5A signaling is associated with several human pathologies such as cancer, fibrosis, and inflammation. Thus, owing to its diverse functions, WNT-5A is a crucial signaling molecule currently under intense investigation with efforts to not only delineate its signaling mechanisms and functions in physiological and pathological conditions, but also to develop strategies for its therapeutic targeting.

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Alprostadil attenuates LPS-induced cardiomyocyte injury by inhibiting the Wnt5a/JNK/NF-κB pathway

Dong

Guan

et al. 2019

Herz

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Background Clinical research has demonstrated that alprostadil has an anti-inflammatory effect; however, to date, its molecular mechanisms remain unclear. This study aimed to examine the anti-inflammatory activity and related mechanisms of alprostadil in lipopolysaccharide (LPS)-treated H9c2 cells. Methods Cell morphology was observed under an inverted light microscope, while cell viability was assessed with the 3‑(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay. Enzyme-linked immunosorbent assays (ELISA) were conducted to study biochemical indicators of cellular damage, such as released lactate dehydrase (LDH) and troponin, and inflammatory cytokine levels including interleukin-1β (IL-1β), IL-6, IL-17, and tumor necrosis factor-α (TNF-α). The mRNA expression levels of Wnt5a, c‑jun N‑terminal kinase (JNK), and nuclear factor kappa B (NF-κB) were further investigated by real-time quantitative polymerase chain reaction (RT-PCR). The effects of alprostadil on the Wnt5a/JNK/NF-κB pathway in H9c2 cells was examined by Western blotting. Results Alprostadil increased the cell viability of LPS-stimulated H9c2 cells, reduced LDH and troponin production, and attenuated IL-1β, IL-6, IL-17, and TNF-α secretion. Moreover, alprostadil reduced the mRNA expression of Wnt5a, JNK, and NF-κB and decreased the expression of Wnt5a, NF-κB, and the ratio of p‑JNK/JNK in H9c2 cells treated with LPS. The siWnt5a or JNK inhibitor SP600125 significantly augmented the inhibitory effects of alprostadil on the Wnt5a/JNK/NF-κB pathway. Conclusion Our results show that alprostadil has anti-inflammatory effects and could attenuate LPS-induced injury in H9c2 cardiomyocytes via the Wnt5a/JNK/NF-κB pathway.

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Wnt5a–Rac1–NF-κB Homeostatic Circuitry Sustains Innate Immune Functions in Macrophages

Cited by 70 publications

References 68 publications

Wnt5a Signaling Promotes Host Defense againstLeishmania donovaniInfection

Wnt5a Signaling Promotes Host Defense againstLeishmania donovaniInfection

WNT-5A: signaling and functions in health and disease

Alprostadil attenuates LPS-induced cardiomyocyte injury by inhibiting the Wnt5a/JNK/NF-κB pathway

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