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用于体内实验的源自外周血单核细胞的猫树突状细胞的生成。

Generation of feline dendritic cells derived from peripheral blood monocytes for in vivo use.

作者信息

Freer Giulia, Matteucci Donatella, Mazzetti Paola, Bozzacco Leonia, Bendinelli Mauro

机构信息

Retrovirus Center and Virology Section, Department of Experimental Pathology, University of Pisa, Via Del Brennero 2, I-56127 Pisa, Italy.

出版信息

Clin Diagn Lab Immunol. 2005 Oct;12(10):1202-8. doi: 10.1128/CDLI.12.10.1202-1208.2005.

Abstract

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.

摘要

树突状细胞(DCs)是专业的抗原呈递细胞,能够启动T细胞并使细胞免疫反应极化。由于Th1型免疫反应与抗击病毒感染的成功相关,DCs的一个有前景的治疗应用是在体外诱导其分化,然后回注到宿主体内,以增强感染动物的免疫反应。本研究旨在开发一种在无外源蛋白条件下培养猫DCs以供体内使用的方案,并研究何种刺激可能最适合诱导其体外成熟并找出成熟的相关因素。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,从外周血单核细胞生成DCs,5天后用脂多糖、人重组肿瘤坏死因子α、聚肌苷酸-聚胞苷酸(poly(I:C))或活化的猫血小板诱导其成熟。48小时后,分析其CD14、CD1a、主要组织相容性复合体II类分子和B7.1的表面表达情况,并同时分析其摄取抗原或启动混合淋巴细胞反应的能力。呈现的结果表明,在自体血浆中培养的猫DCs能够分化,并且在与目前用于其他物种的刺激物类似的刺激下能够成熟。本研究为未来使用通过所述方案获得的DCs进行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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