Velikovsky Carlos A, Cassataro Juliana, Giambartolomei Guillermo H, Goldbaum Fernando A, Estein Silvia, Bowden Raul A, Bruno Laura, Fossati Carlos A, Spitz Moisés
Facultad de Farmacia y Bioquímica Instituto de Estudios de la Inmunidad Humoral, UNICEN, Tandil, Argentina.
Infect Immun. 2002 May;70(5):2507-11. doi: 10.1128/IAI.70.5.2507-2511.2002.
This study was conducted to evaluate the immunogenicity of the Brucella abortus lumazine synthase (BLS) gene cloned into the pcDNA3 plasmid, which is driven by the cytomegalovirus promoter. Injection of plasmid DNA carrying the BLS gene (pcDNA-BLS) into BALB/c mice elicited both humoral and cellular immune responses. Antibodies to the encoded BLS included immunoglobulin G1 (IgG1) IgG2a, IgG2b, IgG3, and IgM isotypes. Animals injected with pcDNA-BLS exhibited a dominance of IgG2a over IgG1. In addition, spleen cells from vaccinated animals produced interleukin-2 and gamma interferon but not IL-10 or IL-4 after in vitro stimulation with recombinant BLS (rBLS), suggesting the induction of a Th1 response. Protection was evaluated by comparing the levels of infection in the spleens of vaccinated mice challenged with B. abortus 544. Immunization with pcDNA-BLS- reduced the bacterial burden relative to those in the control groups. Mice immunized with rBLS produced a significant humoral response but did not show a specific cellular response or any protection from challenge. Altogether, these data suggest that pcDNA-BLS is a good immunogen for the production of humoral and cell-mediated responses in mice and is a candidate for use in future studies of vaccination against brucellosis.
本研究旨在评估克隆到由巨细胞病毒启动子驱动的pcDNA3质粒中的流产布鲁氏菌黄素单核苷酸合酶(BLS)基因的免疫原性。将携带BLS基因的质粒DNA(pcDNA-BLS)注射到BALB/c小鼠体内可引发体液免疫和细胞免疫反应。针对编码的BLS的抗体包括免疫球蛋白G1(IgG1)、IgG2a、IgG2b、IgG3和IgM亚型。注射pcDNA-BLS的动物表现出IgG2a相对于IgG1的优势。此外,接种动物的脾细胞在用重组BLS(rBLS)体外刺激后产生白细胞介素-2和γ干扰素,但不产生IL-10或IL-4,表明诱导了Th1反应。通过比较用流产布鲁氏菌544攻击的接种小鼠脾脏中的感染水平来评估保护作用。与对照组相比,用pcDNA-BLS免疫可降低细菌载量。用rBLS免疫的小鼠产生了显著的体液反应,但未显示出特异性细胞反应或对攻击的任何保护作用。总之,这些数据表明pcDNA-BLS是在小鼠中产生体液和细胞介导反应的良好免疫原,并且是未来布鲁氏菌病疫苗接种研究中使用的候选物。