Flores-Mendoza Lilian, Velazquez Carlos, Bray Jocelyn, Njongmeta Leo, Mwangi Waithaka, Hernández Jesús
Laboratorio de Inmunología, Centro de Investigación en Alimentación y Desarrollo, A.C. Hermosillo, Sonora, México.
Vet Immunol Immunopathol. 2012 Mar 15;146(1):74-80. doi: 10.1016/j.vetimm.2012.01.011. Epub 2012 Jan 20.
The aim of this work was to develop mAbs against porcine CD205 and to conduct a comparative analysis of the CD205 protein expression on lymphoid tissues, monocyte-derived dendritic cells (DCs) and DCs isolated from the porcine skin. To conduct this study, we generated a monoclonal antibody, designated 1.F6F6, against the C-type lectin-like domain-5 of the porcine CD205 and showed that it recognizes a protein band of ∼200 kDa by Western Blot analysis in mesenteric lymph nodes cells. Flow cytometric analysis showed that the mAb 1.F6F6 recognized 28.5%, 28.1% and 34.1% of cells from tonsil, inguinal and mesenteric lymph nodes, respectively, and 6% of cells from thymus. Analysis of monocyte-derived DCs showed that approximately 20% were positive and activation of the cells with LPS increased the positive population to 36%. Analysis of DCs isolated from the porcine skin showed that approximately 70% of the cell population expressed the CD205 receptor. The development of a monoclonal antibody capable of recognizing the CD205 receptor in swine opens up possibilities of applying new strategies for enhancing vaccine efficacy by using the anti-CD205 antibody for DC antigen-targeting to enhance priming of immune responses.
这项工作的目的是开发抗猪CD205的单克隆抗体,并对淋巴组织、单核细胞衍生的树突状细胞(DCs)和从猪皮肤分离的DCs上的CD205蛋白表达进行比较分析。为了开展这项研究,我们制备了一种针对猪CD205的C型凝集素样结构域5的单克隆抗体,命名为1.F6F6,并通过蛋白质印迹分析表明它在肠系膜淋巴结细胞中识别一条约200 kDa的蛋白条带。流式细胞术分析显示,单克隆抗体1.F6F6分别识别扁桃体、腹股沟和肠系膜淋巴结中28.5%、28.1%和34.1%的细胞,以及胸腺中6%的细胞。对单核细胞衍生的DCs的分析表明,约20%为阳性,用脂多糖激活细胞后,阳性群体增加到36%。对从猪皮肤分离的DCs的分析表明,约70%的细胞群体表达CD205受体。能够识别猪CD205受体的单克隆抗体的开发为应用新策略提高疫苗效力开辟了可能性,即使用抗CD205抗体进行DC抗原靶向以增强免疫反应的启动。