Maia Ana Carolina Ribeiro Gomes, Porcino Gabriane Nascimento, Faria-Pinto Priscila, Mendes Túlio Vieira, Antinarelli Luciana Maria Ribeiro, Coimbra Elaine Soares, Reis Alexandre Barbosa, Juliano Luiz, Juliano Maria Aparecida, Marques Marcos José, Vasconcelos Eveline Gomes
Departamento de Bioquímica, Laboratório de Estrutura e Função de Proteínas, Instituto de Ciências Biológicas, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil.
Departamento de Bioquímica, Laboratório de Estrutura e Função de Proteínas, Instituto de Ciências Biológicas, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil.
Vet Parasitol. 2019 Jul;271:38-44. doi: 10.1016/j.vetpar.2019.06.004. Epub 2019 Jun 10.
A nucleoside triphosphate diphosphohydrolase-1 (NTPDase 1) was identified on the surface, flagellum and kinetoplast from L. infantum promastigotes by immunocytochemistry and confocal laser scanning microscopy, using immune sera that recognized specifically the B domain of NTPDase 1 and produced against synthetic peptides (LbB1LJ and LbB2LJ) derived from this domain. The polyclonal antibodies had effective antileishmanial effect, reducing significantly in vitro promastigotes growth (21-25%), an antiproliferative effect also demonstrated by immune sera produced against recombinant r-pot B domain, and two other synthetic peptides (potB1LJ and potB2LJ). In addition, using these biomolecules in ELISA technique, IgG1 and IgG2 subclasses reactivities of either healthy dogs or infected by L. infantum and classified clinically as asymptomatic, oligosymptomatic and symptomatic were tested. Analysis of distinct IgG1 and IgG2 seropositivities patterns suggested antibody subclasses binding epitopes along B domain for protection against infection, indicating this domain as a new tool for prophylactic and immunotherapeutic investigations.
通过免疫细胞化学和共聚焦激光扫描显微镜技术，利用特异性识别核苷三磷酸二磷酸水解酶-1（NTPDase 1）B结构域的免疫血清，并针对源自该结构域的合成肽（LbB1LJ和LbB2LJ）制备的免疫血清，在婴儿利什曼原虫前鞭毛体的表面、鞭毛和动基体上鉴定出了NTPDase 1。多克隆抗体具有有效的抗利什曼原虫作用，显著降低了体外前鞭毛体的生长（21%-25%），针对重组r-pot B结构域以及另外两种合成肽（potB1LJ和potB2LJ）产生的免疫血清也显示出抗增殖作用。此外，利用这些生物分子采用酶联免疫吸附测定（ELISA）技术，检测了健康犬或感染婴儿利什曼原虫且临床分类为无症状、轻度症状和有症状的犬的IgG1和IgG2亚类反应性。对不同的IgG1和IgG2血清阳性模式的分析表明，抗体亚类沿着B结构域结合表位以预防感染，表明该结构域是预防性和免疫治疗研究的新工具。
