CD3ε of a pan T cell marker involved in mouse Aspergillus fumigatus keratitis

AIM:To explore whether CD3ε is involved in the adaptive immunity of Aspergillus fumigatus(A.fumigatus)keratitis in mice and the role of innate and adaptive immunity in it.METHODS:Mice models of A.fumigatus keratitis were established by intra-stromal injection and corneal epithelial scratching.Subconjunctival injections of natamycin,wedelolactone,LOX-1 inhibitor(poly I)or Dectin-1 inhibitor(laminarin)were used to treat mice with A.fumigatus keratitis.Mice were pretreated by intraperitoneal injection of anti-mouse CD3ε.We observed the corneal infection of mice under the slit lamp microscope and made a clinical score.The protein expression of CD3ε and interleukin-10(IL-10)was determined by Western blotting.RESULTS:With the disease progresses,the degree of corneal opacity and edema augmented.In the intrastromal injection models,CD3εprotein expression began to increase significantly on the 2^(nd) day.However,in the scraping epithelial method models,CD3ε only began to increase on the 3^(rd) day.After natamycin treatment,the degree of corneal inflammation in mice was significantly attenuated on the 3^(rd) day.After wedelolactone treatment,the severity of keratitis worsened.And the amount of CD3ε protein was also reduced,compared with the control group.By inhibiting LOX-1 and Dectin-1,there was no significant difference in CD3ε production compared with the control group.After inhibiting CD3ε,corneal ulcer area and clinical score increased,and IL-10 expression was downregulated.CONCLUSION:As a pan T cell marker,CD3ε participate in the adaptive immunity of A.fumigatus keratitis in mice.In our mice models,the corneas will enter the adaptive immune stage faster.By regulating IL-10,CD3ε exerts antiinflammatory and repairs effects in the adaptive immune stage.

Thymol Protects against Aspergillus Fumigatus Keratitis by Inhibiting the LOX-1/IL-1βSignaling Pathway

Objective To explore the anti-inflammatory effects and mechanisms of action of thymol in Aspergillus fumigatus(A.fumigatus)keratitis.Methods The minimum inhibitory concentration of thymol against A.fumigatus was detected.To characterize the anti-inflammatory effects of thymol,mouse corneas and human corneal epithelial cells were pretreated with thymol or dimethyl sulfoxide(DMSO)before infection with A.fumigatus spores.Slit-lamp microscopy,immunohistochemistry,myeloperoxidase detection,quantitative real-time polymerase chain reaction,and Western blotting were used to assess infection.Neutrophil and macrophage recruitment,in addition to the secretion of LOX-1 and IL-1β,were quantified to evaluate the relative contribution of thymol to the inflammatory response.Results We confirmed that the growth of A.fumigatus was directly inhibited by thymol.In contrast with the DMSO group,there was a lower degree of inflammation in the mouse corneas of the thymol-pretreated group.This was characterized by significantly lower clinical scores,less inflammatory cell infiltration,and lower expression of LOX-1 and IL-1β.Similarly,in vitro experiments indicated that the production of LOX-1 and IL-1βwas significantly inhibited after thymol treatment,in contrast with the DMSO-pretreated group.Conclusion Our findings demonstrate that thymol exerted a direct fungistatic activity on A.fumigatus.Furthermore,thymol played a protective role in fungal keratitis by inhibiting LOX-1/IL-1βsignaling pathway and reducing the recruitment of neutrophils and macrophages.

CCPG1 involved in corneal Aspergillus fumigatus infection

AIM: To investigate whether non-canonical autophagy transport receptor cell cycle progression 1(CCPG1) is involved in the corneal antifungal immune response.METHODS: Human corneal epithelial cells(HCECs) and human myeloid leukemia mononuclear cells(THP-1) macrophages stimulated by Aspergil us fumigatus(A. fumigatus) were used as cell models. The expression of CCPG1 m RNA was detected by q RT-PCR. Western blot was used to determine the protein expression of CCPG1 and interleukin-1β(IL-1β). The dectin-1 neutralizing antibody was used to detect the association between dectin-1 and CCPG1. Immunofluorescence was used to observe the colocalization of CCPG1 and C-type lectin-like receptor-1(CLEC-1) in THP-1 macrophages.RESULTS: The expression of CCPG1 started to increase at 4 h after infection and increased in a time-dependent manner in HCECs and THP-1 macrophages. With dectin-1 neutralizing antibody pretreatment, the expression of IL-1β was down-regulated. CCPG1 up-regulation in response to A. fumigatus infection was independent of dectin-1. Immunofluorescence showed the colocalization of CCPG1 and CLEC-1 in THP-1 macrophages.CONCLUSION: As a specific autophagy protein of noncanonical autophagy pathway, CCPG1 is involved in corneal infection with A. fumigatus.

CD23 mediated the induction of pro-inflammatory cytokines Interleukin-1 beta and tumor necrosis factors-alpha in Aspergillus fumigatus keratitis

To the Editor:Fungal keratitis is primarily caused by pathogenic fungi species,such as Fusarium solani and Aspergillus fumigatus.The incidence of A.fumigatus has been increased worldwide.[1]Although new therapies have been used for fungal keratitis,the efficacy of antifungal drugs remains unsatisfactory at present.[2]Fungal keratitis is still a challenge for ophthalmologists because of its difficult and delayed diagnosis,as well as the lack of effective drugs and treatment.

Structure Engineering and Electronic Modulation of Transition Metal Interstitial Compounds for Electrocatalytic Water Splitting

CONSPECTUS:Hydrogen is deemed as an ideal energy carrier because of its high energy density and clean nature.Water electrolysis is fairly competitive for hydrogen production due to the conversion of renewable electricity to high-purity H2 with no carbon emission,in comparison with traditional industrial technology.However,the large-scale application is hampered by high cost partially from the use of noble metal-based catalysts to promote the kinetics of hydrogen and oxygen evolution reactions.Developing cost-efficient transition metal-based electrocatalysts,therefore,is a hopeful prospect,because they can provide dorbital lone-pair electrons or empty d-orbitals for adsorbing different intermediates(such as H*,OH*,O*,and OOH*).As compared to transition metals and their oxides,transition metal interstitial compounds(TMICs)formed by inserting C,N,and P atoms into the interstitial sites of parent metals hold distinct advantages in their Pt-like electronic structure,high conductivity,and superior chemical stability over a wide pH range,beneficial to overcoming the high energy consumption faced by alkaline water electrolysis and the intractable stability issue of acid water electrolysis.Nevertheless,the major drawbacks are large size,high density,and sluggish ionic kinetics,resulting in ordinary electrochemical activity and low mass efficiency.Electrocatalytic performance is dominated by the intrinsic activity,the number of accessible active sites,and the capacity of charge and mass transfer.Engineering the micronano structure(small-size particles,porous structure,and ultrathin nanosheet)can expose more catalytical active sites and facilitate mass transport and gas diffusion.Meanwhile,modulating the electronic structure can optimize the adsorption energy of the intermediates to boost the intrinsic activity.Apparently,synergistic modulation of the micronano structure and electronic structure of TMICs is expected to achieve the multiobjective optimization for targeting the highly effective catalysts.In this Account,we summarize our recent efforts in the designed synthesis and structure engineering of TMICs by utilizing polyoxometalates(POMs)as metal precursors and the associated electronic modulation strategies to advance the electrocatalytic performance toward HER and OER.We start with a brief summary of the HER and OER mechanisms,which play crucial roles in the elaborate design of the relevant electrocatalysts.The advantages and disadvantages of TMICs for water electrolysis are pointed out,apart from the opportunities offered by POMs for constructing novel TMICs from size,component,and interface structure.Several efficient strategies for performance enhancement are proposed including reducing the size to expose more accessible active sites,constructing heterojunctions to provide highly active interfaces,doping heteroatoms to regulate the binding energy of intermediates,and creating pores to accelerate mass transfer,etc.Accordingly,the TMICs with controllable size and well-defined structure are highlighted,in which the positive role of tailoring the micronano structure and electronic structure on enhancing the catalytic efficiency is confirmed.Furthermore,paired electrocatalysis by using hydrogen and oxygen active species from water is proposed to produce value-added chemicals and reduce energy consumption.Finally,the remaining challenges,opportunities,and future development directions of TMICs-based materials toward electrocatalytic energy conversion are discussed.