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中文摘要：
肺泡巨噬細(xì)胞 （AM） 是專門的組織駐留巨噬細(xì)胞，可在過敏性炎癥和哮喘中協(xié)調(diào)免疫反應(yīng)。然而，是什么信號指示 AM 與其他免疫細(xì)胞進(jìn)行串?dāng)_仍不清楚。在這里，我們報(bào)道了自分泌運(yùn)動因子受體 （AMFR），一種內(nèi)質(zhì)網(wǎng)駐留的 E3 泛素連接酶，在哮喘的 AM 中上調(diào)，對這種情況至關(guān)重要。AMFR 缺乏顯著降低過敏誘導(dǎo)的輔助性 T 細(xì)胞 2 （Th2） 和嗜酸性粒細(xì)胞炎癥，AM 中粒細(xì)胞-巨噬細(xì)胞集落刺激因子 （GM-CSF） 的產(chǎn)生較少。從機(jī)制上講，在胸腺基質(zhì)淋巴細(xì)胞生成素 （TSLP） 刺激后，AMFR 與細(xì)胞因子誘導(dǎo)的含 SH2 的蛋白 （CIS） 直接相關(guān)，誘導(dǎo) CIS 的 Lys48 連接的多泛素化的泛素化，從而阻斷了 CIS 對信號轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄激活因子 5 （STAT5） 磷酸化和 AM 下游途徑激活的抑制作用?？傊?，我們的結(jié)果表明，AMFR 通過調(diào)節(jié) AM 功能在促進(jìn)哮喘炎癥中起關(guān)鍵作用，并可能成為哮喘治療的新潛在藥物靶點(diǎn)。
英文摘要：
Alveolar macrophages (AMs) are specialized tissue-resident macrophages that orchestrate the immune response in allergic inflammation and asthma. However, what signals direct AMs to cross talk with other immune cells remains unclear. Here, we report that autocrine motility factor receptor (AMFR), an endoplasmic reticulum–resident E3 ubiquitin ligase, is upregulated in AMs of asthma and is critical for this condition. AMFR deficiency significantly decreased allergy-induced T helper 2 (Th2) and eosinophilic inflammation, with less granulocyte-macrophage colony-stimulating factor (GM-CSF) production in AMs. Mechanistically, following thymic stromal lymphopoietin (TSLP) stimulation, AMFR associated directly with cytokine-inducible SH2-containing protein (CIS), induced the ubiquitination of Lys48-linked polyubiquitination of CIS, and consequently blocked the inhibitory effect of CIS on signal transducer and activator of transcription 5 (STAT5) phosphorylation and the downstream pathway activation in AMs. In conclusion, our results demonstrate that AMFR serves a crucial role in promoting inflammation in asthma through regulating AM function, and may emerge as a new potential drug target for asthma therapy.
論文信息：
論文題目： AMFR drives allergic asthma development by promoting alveolar macrophage–derived GM-CSF production
期刊名稱：JEM- J Exp Med
時(shí)間期卷： (2022) 219 (5): e20211828
在線時(shí)間：2022年3月25日
DOI： doi.org/10.1084/jem.20211828
Liposoma巨噬細(xì)胞清除劑氯膦酸鹽脂質(zhì)體Clodronate Liposomes見刊于JEM：
Liposoma巨噬細(xì)胞清除劑氯膦酸鹽脂質(zhì)體Clodronate Liposomes的材料和方法：
JEM期刊率膦酸鹽脂質(zhì)體清除肺泡巨噬細(xì)胞專業(yè)論文：肺泡巨噬細(xì)胞清除解決方案
OVA (A5503), papain (76216), chitin (C9752), collagenase D (11088866001), and DNase I (10104159001) were obtained from Sigma-Aldrich. The Imject Alum adjuvant (77161) and ER-TrackerTM Blue-White DPX (E12353) were purchased from Thermo Fisher Scientific. Clodronate liposomes (CP-005-005) were purchased from Liposoma. The recombinant murine (555-TS) and human (1398-TS) TSLP cytokines were purchased from R&D. Recombinant murine GM-CSF (315-03) and M-CSF (315-02) were from PeproTech. The anti-AMFR (ab76841) antibody was obtained from Abcam. The anti-CIS antibody (sc-166326) was obtained from Santa Cruz Biotechnology. The anti-CD68 antibody (14-0681-80) was purchased from Invitrogen Thermo Fisher Scientific. Antibodies for Myc-Tag (2272S and 2276S), Flag-Tag (14793S), HA-Tag (3724S), β-actin (8457S), Ub (3936S), STAT5 (94205S), phospho-STAT5 (9351L), phospho-JAK1 (74129T), JAK1 (3344T), phospho-JAK2 (8082T), JAK2, (3230T), SOCS1 (3950T), SOCS2 (2779P), SOCS3 (2932P), Alexa Fluor 594 anti-mouse IgG (8890S), and Alexa Fluor 488 anti-rabbit IgG (4412S) were obtained from Cell Signaling Technology. The secondary antibodies peroxidase-conjugated anti-rabbit (111-035-003) and anti-mouse (115-035-003) were purchased from Jackson ImmunoResearch Laboratories. The flow cytometry antibodies, including APC anti-mouse CD11c (117310), FITC anti-mouse Siglec-F (155504), PE anti-mouse Siglec-F (155506), APC anti-mouse/human CD11b (101212), PE/Cyanine7 anti-mouse CD45 (103114), PerCP/Cyanine5.5 anti-mouse CD64 (139307), APC/Fire 750 anti-mouse Ly-6G (127652), Brilliant Violet 650 anti-mouse F4/80 (123149), Brilliant Violet 421 anti-mouse/human CD11b (101235), FITC anti-mouse I-A/I-E (MHC class II; 107605), PE anti-human GM-CSF (502305), and PE/Cyanine7 anti-mouse GM-CSF (505411), were from BioLegend. The BCA protein assay kit (P0012S) and DAPI (C1002) were obtained from Beyotime.
Adoptive transfer of AMs was performed as previously reported (Miki et al., 2021; Qian et al., 2015). For in vivo deletion of macrophages in lung tissues, mice were sensitized with OVA as described above and treated with 40 μl of clodronate liposome i.t. for two successive days (days 18 and 19). For the AM adoptive transfer study, AMs derived from WT or AMFR knockout mice were then transferred by i.t. injection into the lungs of clodronate liposome-treated and OVA-sensitized WT mice at a density of 5 × 105 cells/mouse (40 μl) on day 20. 24 h after AM delivery, the mice were i.t. challenged with OVA for three days (days 21, 22, and 23). On day 25, the mice were sacrificed to analyze allergic asthmatic inflammation (Fig. 3 A).
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