α-半乳糖神经酰胺通过干扰素-γ依赖性诱导一氧化氮合酶和CD95来抑制小鼠嗜酸性粒细胞的产生。

α-Galactosylceramide suppresses murine eosinophil production through interferon-γ-dependent induction of NO synthase and CD95.

作者信息

Gaspar-Elsas Maria Ignez, Queto Túlio, Masid-de-Brito Daniela, Vieira Bruno Marques, de Luca Bianca, Cunha Fernando Queiroz, Xavier-Elsas Pedro

机构信息

Department of Pediatrics, Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira, FIOCRUZ, Rio de Janeiro, Brazil.

Department of Immunology, Instituto de Microbiologia Prof. Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.

出版信息

Br J Pharmacol. 2015 Jul;172(13):3313-25. doi: 10.1111/bph.13126. Epub 2015 Apr 24.

DOI:10.1111/bph.13126

PMID:25752588

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4500368/

Abstract

BACKGROUND AND PURPOSE

α-Galactosylceramide (α-GalCer), a pleiotropic immunomodulator with therapeutic potential in neoplastic, autoimmune and allergic diseases, activates invariant natural killer T-cells throughCD1-restricted receptors for α-GalCer on antigen-presenting cells, inducing cytokine secretion. However the haemopoietic effects of α-GalCer remain little explored.

EXPERIMENTAL APPROACH

α-GalCer-induced modulation of eosinophil production in IL-5-stimulated bone marrow cultures was examined in wild-type (BALB/c, C57BL/6) mice and their mutants lacking CD1, inducible NOS (iNOS), CD95 and IFN-γ, along with the effects of lymphocytes; IFN-γ; caspase and iNOS inhibitors; non-steroidal anti-inflammatory drugs (NSAIDs) and LTD4 ; and dexamethasone.

KEY RESULTS

α-GalCer (10(-6) -10(-8) M) suppressed IL-5-stimulated eosinopoiesis by inducing apoptosis. α-GalCer pretreatment in vivo (100 μg·kg(-1) , i.v.) suppressed colony formation by GM-CSF-stimulated bone marrow progenitors in semi-solid cultures. α-GalCer and dexamethasone synergistically promoted eosinophil maturation. Suppression of eosinophil production by α-GalCer was prevented by aminoguanidine and was undetectable in bone marrow lacking iNOS, CD95, CD28; or CD1d. Separation on Percoll gradients and depletion of CD3+ cells made bone marrow precursors unresponsive to α-GalCer. Responsiveness was restored with splenic lymphocytes. Experiments with (i) IFN-γ-deficient bone marrow, alone or co-cultured with spleen T-cells from wild-type, but not from CD1d-deficient, donors; (ii) IFN-γ neutralization; and (iii) recombinant IFN-γ, showed that these effects of α-GalCer were mediated by IFN-γ. Effects of α-GalCer on eosinophil production were blocked by LTD4 and NSAIDs.

CONCLUSIONS AND IMPLICATIONS

α-GalCer activation of IFN-γ-secreting, CD1d-restricted lymphocytes induced iNOS-CD95-dependent apoptosis in developing eosinophils. This pathway is initiated by endogenous regulatory lymphocytes, antagonised by LTD4 , NSAIDs and aminoguanidine, and modified by dexamethasone.

摘要

背景与目的

α-半乳糖神经酰胺（α-GalCer）是一种具有多效性的免疫调节剂，在肿瘤、自身免疫性疾病和过敏性疾病中具有治疗潜力，它通过抗原呈递细胞上α-GalCer的CD1限制性受体激活不变自然杀伤T细胞，诱导细胞因子分泌。然而，α-GalCer对造血的影响仍鲜有研究。

实验方法

在野生型（BALB/c、C57BL/6）小鼠及其缺乏CD1、诱导型一氧化氮合酶（iNOS）、CD95和干扰素-γ（IFN-γ）的突变体中，检测α-GalCer对白细胞介素-5（IL-5）刺激的骨髓培养物中嗜酸性粒细胞生成的调节作用，以及淋巴细胞、IFN-γ、半胱天冬酶和iNOS抑制剂、非甾体抗炎药（NSAIDs）和白三烯D4（LTD4）以及地塞米松的影响。

主要结果

α-GalCer（10⁻⁶ - 10⁻⁸ M）通过诱导凋亡抑制IL-5刺激的嗜酸性粒细胞生成。体内α-GalCer预处理（100 μg·kg⁻¹，静脉注射）抑制了半固体培养物中粒细胞-巨噬细胞集落刺激因子（GM-CSF）刺激的骨髓祖细胞的集落形成。α-GalCer和地塞米松协同促进嗜酸性粒细胞成熟。氨基胍可阻止α-GalCer对嗜酸性粒细胞生成的抑制作用，在缺乏iNOS、CD95、CD28或CD1d的骨髓中未检测到这种抑制作用。通过Percoll梯度分离和去除CD3⁺细胞使骨髓前体细胞对α-GalCer无反应。脾淋巴细胞可恢复其反应性。使用（i）IFN-γ缺陷型骨髓，单独或与来自野生型而非CD1d缺陷型供体的脾T细胞共培养；（ii）IFN-γ中和；以及（iii）重组IFN-γ进行的实验表明，α-GalCer的这些作用是由IFN-γ介导的。LTD4和NSAIDs可阻断α-GalCer对嗜酸性粒细胞生成的影响。