Instituto de Virología e Innovaciones Tecnológicas (IVIT, INTA-CONICET) Hurlingham, Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
Instituto de Virología e Innovaciones Tecnológicas (IVIT, INTA-CONICET) Hurlingham, Buenos Aires, Argentina.
Vaccine. 2021 Feb 5;39(6):1007-1017. doi: 10.1016/j.vaccine.2020.11.071. Epub 2021 Jan 11.
DNA vaccines are capable of inducing humoral and cellular immunity, and are important to control bovine herpesvirus 1 (BoHV-1), an agent of the bovine respiratory disease complex. In previous work, a DNA plasmid that encodes a secreted form of BoHV-1 glycoprotein D (pCIgD) together with commercial adjuvants provided partial protection against viral challenge of bovines. In this work, we evaluate new molecules that could potentiate the DNA vaccine. We show that a plasmid encoding a soluble CD40 ligand (CD40L) and the adjuvant Montanide™ GEL01 (GEL01) activate in vitro bovine afferent lymph dendritic cells (ALDCs). CD40L is a co-stimulating molecule, expressed transiently on activated CD4+ T cells and, to a lesser extent, on activated B cells and platelets. The interaction with its receptor, CD40, exerts effects on the presenting cells, triggering responses in the immune system. GEL01 was designed to improve transfection of DNA vaccines. We vaccinated cattle with: pCIgD; pCIgD-GEL01; pCIgD with GEL01 and CD40L plasmid (named pCIgD-CD40L-GEL01) or with pCIneo vaccines. The results show that CD40L plasmid with GEL01 improved the pCIgD DNA vaccine, increasing anti-BoHV-1 total IgGs, IgG1, IgG2 subclasses, and neutralizing antibodies in serum. After viral challenge, bovines vaccinated with pCIgD-GEL01-CD40L showed a significant decrease in viral excretion and clinical score. On the other hand, 80% of animals in group pCIgD-GEL01-CD40L presented specific anti-BoHV-1 IgG1 antibodies in nasal swabs. In addition, PBMCs from pCIgD-CD40L-GEL01 had the highest percentage of animals with a positive lymphoproliferative response against the virus and significant differences in the secretion of IFNγ and IL-4 by mononuclear cells, indicating the stimulation of the cellular immune response. Overall, the results demonstrate that a plasmid expressing CD40L associated with the adjuvant GEL01 improves the efficacy of a DNA vaccine against BoHV-1.
DNA 疫苗能够诱导体液和细胞免疫，对于控制牛疱疹病毒 1（BoHV-1）至关重要，BoHV-1 是牛呼吸道疾病复合体的病原体。在之前的工作中，一种编码 BoHV-1 糖蛋白 D 分泌形式的 DNA 质粒与商业佐剂一起提供了对牛病毒攻击的部分保护。在这项工作中，我们评估了可能增强 DNA 疫苗效力的新分子。我们表明，一种编码可溶性 CD40 配体（CD40L）的质粒和佐剂 Montanide™ GEL01（GEL01）能够在体外激活牛传入淋巴树突状细胞（ALDCs）。CD40L 是一种共刺激分子，短暂表达于活化的 CD4+T 细胞上，在一定程度上也表达于活化的 B 细胞和血小板上。它与受体 CD40 的相互作用对呈递细胞产生影响，触发免疫系统的反应。GEL01 的设计目的是提高 DNA 疫苗的转染效率。我们用以下疫苗对牛进行了免疫接种：pCIgD；pCIgD-GEL01；pCIgD 与 GEL01 和 CD40L 质粒（命名为 pCIgD-CD40L-GEL01）或 pCIneo 疫苗。结果表明，与 GEL01 联合使用 CD40L 质粒可增强 pCIgD DNA 疫苗的效果，增加血清中抗 BoHV-1 的总 IgG、IgG1、IgG2 亚类和中和抗体。在病毒攻击后，接种 pCIgD-GEL01-CD40L 的牛的病毒排出量和临床评分显著降低。另一方面，pCIgD-GEL01-CD40L 组 80%的动物在鼻拭子中呈现出针对 BoHV-1 的特异性 IgG1 抗体。此外，pCIgD-CD40L-GEL01 的 PBMC 中，有最高比例的动物对病毒产生了阳性的淋巴增殖反应，且单核细胞分泌 IFNγ 和 IL-4 的差异显著，表明细胞免疫反应得到了刺激。总体而言，结果表明，表达 CD40L 的质粒与佐剂 GEL01 联合使用可提高针对 BoHV-1 的 DNA 疫苗的效力。
