巨噬細(xì)胞的酸性神經(jīng)酰胺酶將單純皰疹病毒困在多囊泡體中，并防止嚴(yán)重疾病

中文摘要：

巨噬細(xì)胞在感染單純皰疹病毒1型（HSV-1）期間具有重要的保護(hù)功能。然而，限制病毒傳播并防止嚴(yán)重疾病的分子機(jī)制尚不清楚。在這里，我們顯示巨噬細(xì)胞通過內(nèi)吞作用攝取HSV-1，并將病毒顆粒運(yùn)送到多泡體（MVBs）中。在MVBs中，酸性神經(jīng)酰胺酶（aCDase）將神經(jīng)酰胺轉(zhuǎn)化為鞘氨醇，并增加富含鞘氨醇的內(nèi)腔囊泡（ILVs）的形成。一旦HSV-1顆粒到達(dá)MVBs，富含鞘氨醇的ILVs會(huì)與HSV-1顆粒結(jié)合，從而限制其與限制性內(nèi)體膜的融合，并阻止細(xì)胞感染。在巨噬細(xì)胞培養(yǎng)中缺乏aCDase或在Asah1?/? 小鼠中會(huì)導(dǎo)致HSV-1的復(fù)制，Asah1?/? 小鼠在全身或陰道內(nèi)接種后很快死亡。對(duì)巨噬細(xì)胞進(jìn)行增強(qiáng)鞘氨醇的化合物處理可阻止HSV-1的傳播，提示該途徑具有潛在的治療價(jià)值?？傊?，aCDase將ILVs加載鞘氨醇，從而阻止HSV-1衣殼進(jìn)入胞質(zhì)。

英文摘要：

Macrophages have important protective functions during infection with herpes simplex virus type 1 (HSV-1). However, molecular mechanisms that restrict viral propagation and protect from severe disease are unclear. Here we show that macrophages take up HSV-1 via endocytosis and transport the virions into multivesicular bodies (MVBs). In MVBs, acid ceramidase (aCDase) converts ceramide into sphingosine and increases the formation of sphingosine-rich intraluminal vesicles (ILVs). Once HSV-1 particles reach MVBs, sphingosine-rich ILVs bind to HSV-1 particles, which restricts fusion with the limiting endosomal membrane and prevents cellular infection. Lack of aCDase in macrophage cultures or in Asah1?/? mice results in replication of HSV-1 and Asah1?/? mice die soon after systemic or intravaginal inoculation. The treatment of macrophages with sphingosine enhancing compounds blocks HSV-1 propagation, suggesting a therapeutic potential of this pathway. In conclusion, aCDase loads ILVs with sphingosine, which prevents HSV-1 capsids from penetrating into the cytosol.

論文信息：

論文題目：Acid ceramidase of macrophages traps herpes simplex virus in multivesicular bodies and protects from severe disease

期刊名稱：Nature Communications

時(shí)間期卷：11, Article number: 1338(2020)

在線時(shí)間：2020年3月12日

DOI： doi.org/10.1038/s41467-020-15072-8

產(chǎn)品信息：

貨號(hào)：CP-005-005

規(guī)格：5ml+5ml

品牌：Liposoma

產(chǎn)地：荷蘭

名稱：Clodronate Liposomes& Control Liposomes

辦事處：Target Technology(靶點(diǎn)科技)

Clodronate Liposomes氯膦酸鹽脂質(zhì)體清除病毒模型中巨噬細(xì)胞 ，荷蘭Liposoma巨噬細(xì)胞清除劑ClodronateLiposomes見刊于Nature Communications：巨噬細(xì)胞的酸性神經(jīng)酰胺酶將單純皰疹病毒困在多囊泡體中，并防止嚴(yán)重疾病。

Liposoma巨噬細(xì)胞清除劑Clodronate Liposomes氯膦酸二鈉脂質(zhì)體清除腫瘤相關(guān)巨噬細(xì)胞的材料和方法：

Reagents

D-erythro-sphingosine (C18, 860490) and D-Galactosyl-β1-1’-N-Nervonoyl-D-erythro-sphingosine (C24:1, β-D-Galactosyl Ceramide, 110759), were obtained from Avanti Polar Lipids (Alabaster, AL, USA). Glycoceramidase (E9030-100MUN), sphingomyelinase (S8633-25UN), Cy3-linked DBCO (777366-1?mg) and DAPI (D9542) were purchased from Millipore Sigma (Darmstadt, Germany). Recombinant mouse Asah2 (3558-AH) was purchased from R&D Systems (Minneapolis, MN, USA). The sphingosine kinase inhibitor SKI-II (CAS # 312636-16-1; Cat # 2097) was obtained from Tocris Bioscience (Bristol, United Kingdom) and tamoxifen (CAS # 10540-29-1, Cat # T5648-5G) and cornoil (CAS # 8001-30-7, Cat # C8267-500ML) from Millipore Sigma. Clodronate and control liposomes were purchased from Liposoma (CP-005-005, Amsterdam, Netherlands). Clickable ω-azido-sphingosine ((2S,3R,E)-2-amino-18-azidooctadec-4-ene-1,3-diol) for click chemistry42 staining was synthesized in-house by Julian Fink.

Depletion of macrophages

For macrophage-depleted mouse experiments, mice were treated intravenously with 50?mg/kg clodronate to deplete tissue resident macrophages. After 3 days, mice were ready to be used in experiments.

材料和方法文獻(xiàn)截圖：