技術(shù)文章
Vimentin (波形蛋白) 是一種在間充質(zhì)細(xì)胞中廣泛表達(dá)的 III 型中間絲 (intermediate filament, IF) 蛋白。它在細(xì)胞骨架的形成、細(xì)胞的形狀維持、細(xì)胞的遷移、細(xì)胞信號傳導(dǎo)以及細(xì)胞應(yīng)激反應(yīng)中發(fā)揮著重要作用。它通過側(cè)向或末端方式附著于細(xì)胞核、內(nèi)質(zhì)網(wǎng)和線粒體,參與調(diào)控細(xì)胞定向運動、取向排列、細(xì)胞片層組織形成,并在細(xì)胞遷移前沿維持高爾基體極性。此外,波形蛋白通過細(xì)胞接觸依賴機制保護(hù)SCRIB蛋白免受蛋白酶體降解,并促進(jìn)其定位于中間絲結(jié)構(gòu)
Vimentin結(jié)構(gòu). Schematic representation of a vimentin filament from a monomer to its assembly into a mature filament. The vimentin monomer is composed of a central a-helical domain flanked by an unstructured head and tail domain. On the central domain, vimentin possesses a rod II domain with lectin-like properties.
重要發(fā)現(xiàn):Vimentin高表達(dá)(VIM+)巨噬細(xì)胞通過分泌IL-1β激活Treg細(xì)胞的免疫抑制功能
復(fù)旦大學(xué)的王紅陽/陳磊教授團隊于2024年9月在Nature Cancer (IF:23.5)上發(fā)表了題為“Spatial single-cell protein landscape reveals vimentinhigh macrophages as immune-suppressive in the microenvironment of hepatocellular carcinoma"的研究性文章。該文繪制了HCC單細(xì)胞空間蛋白圖譜,解析了HCC腫瘤間空間異質(zhì)性,建立了預(yù)后相關(guān)的干細(xì)胞癌(HCC)空間分型(Spatial Pattern),并整合多組學(xué)和體外實驗揭示了Vimentin高表達(dá)的巨噬細(xì)胞與Treg細(xì)胞的空間互作促進(jìn)HCC的疾病進(jìn)展。
該項研究通過開展了單細(xì)胞測序和空間轉(zhuǎn)錄組分析,揭示了VIM+巨噬細(xì)胞的基因表達(dá)特征,發(fā)現(xiàn)了VIM+巨噬細(xì)胞和Treg之間基因表達(dá)的高度關(guān)聯(lián)性,提示了VIM+巨噬細(xì)胞可能作用于Treg,從而增強免疫抑制。配受體分析進(jìn)一步揭示了VIM+巨噬細(xì)胞或許通過白介素-1β(IL-1β)和干擾素-γ(IFN-γ)參與對Treg細(xì)胞的調(diào)控。隨后,作者利用流式細(xì)胞儀分離了VIM+和VIM低表達(dá)的巨噬細(xì)胞,并檢測了其中IL-1β的表達(dá)水平。結(jié)果表明,在不同類型的巨噬細(xì)胞中,VIM+的巨噬細(xì)胞顯著高表達(dá)IL-1β,細(xì)胞共培養(yǎng)實驗發(fā)現(xiàn)了VIM+巨噬細(xì)胞促進(jìn)Treg分泌IL-10,而加入IL-1β抗體可顯著緩解作用效果。
斯達(dá)特S-RMab®?
Vimentin Recombinant Rabbit mAb (SDT-029-120)?(貨號:S0B2254)抗體是您Vimentin 研究中的有力幫手。
數(shù)據(jù)驗證:
WB免疫印記
WB result of Vimentin Rabbit mAb
Primary antibody: Vimentin Rabbit mAb at 1/500 dilution
Lane 1: Daudi whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Lane 3: A549 whole cell lysate 20 µg
Negative control: Daudi whole cell lysate
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 54 kDa
Observed MW: 57 kDa
IHC 免疫組化
斯達(dá)特S-RMab® 系列抗體賦能生命科學(xué)和病理研究
免疫細(xì)胞化學(xué)
ICC shows positive staining in HeLa cells. Anti-Vimentin antibody was used at 1/250 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue).Counterstain with tubulin (Red).
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明星指標(biāo):高表達(dá)vimentin的巨噬細(xì)胞在肝細(xì)胞癌免疫抑制中的關(guān)鍵作用
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