来源于外周血单核细胞的猫树突状细胞的生成用于体内使用。

Giulia Freer, Donatella Matteucci, Paola Mazzetti, Leonia Bozzacco, Mauro Bendinelli
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引用次数: 22

摘要

树突状细胞(dc)是一种专业的抗原呈递细胞,可以启动T细胞并极化细胞免疫反应。由于th1型免疫反应与成功对抗病毒感染有关,因此dc的一个有希望的治疗应用将是它们在体外分化并注射回宿主以增强受感染动物的免疫反应。本研究旨在制定一种在缺乏外源蛋白的情况下培养猫树突状细胞的方案,以供其在体内使用,并研究在体外诱导其成熟的最合适刺激是什么,并找到成熟的相关因素。我们在猫白细胞介素-4和粒细胞-巨噬细胞集落刺激因子存在的情况下,从外周血单核细胞中生成DCs, 5天后用脂多糖、人重组肿瘤坏死因子α、聚(I:C)或活化的猫血小板诱导其成熟。48 h后,分析其CD14、CD1a、主要组织相容性复合体II类和B7.1表面表达,同时分析其摄取抗原或引发混合白细胞反应的能力。结果表明,在自体血浆中培养的猫树突状细胞能够分化,并且能够在类似于目前用于其他物种的刺激下成熟。目前的工作为未来使用该方案获得的dc用于猫免疫缺陷病毒感染猫的体内疫苗接种和免疫治疗奠定了基础。
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Generation of feline dendritic cells derived from peripheral blood monocytes for in vivo use.

Dendritic cells (DCs) are professional antigen-presenting cells that can prime T cells and polarize the cellular immune response. Because Th1-type immune responses have been connected to success in combating viral infection, a promising therapeutic application of DCs would be their differentiation in vitro and injection back into the host to boost an immune response in infected animals. This study was aimed both at developing a protocol to cultivate feline DCs in the absence of exogenous proteins for their use in vivo and at investigating what might be the most appropriate stimulus to induce their maturation in vitro and finding correlates of maturation. We generated DCs from peripheral blood monocytes in the presence of feline interleukin-4 and granulocyte-macrophage colony stimulating factor, and after 5 days their maturation was induced with either lipopolysaccharide, human recombinant tumor necrosis factor alpha, poly(I:C), or activated feline platelets. After 48 h, their CD14, CD1a, major histocompatibility complex class II, and B7.1 surface expression was analyzed in parallel with their ability to uptake antigen or prime a mixed leukocyte reaction. The results presented show that feline DCs cultured in autologous plasma differentiate and are able to mature in the presence of stimuli similar to the ones currently used for other species. The present work sets the grounds for future use of DCs obtained by the protocol described for in vivo vaccination and immunotherapy of feline immunodeficiency virus-infected cats.

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