Fernandes Andrea Claudia Bekner Silva, Pedroso Raíssa Bocchi, de Mello Tatiane França Perles, Donatti Lucélia, Venazzi Eneide Aparecida Sabaini, Demarchi Izabel Galhardo, Aristides Sandra Mara Alessi, Lonardoni Maria Valdrinez Campana, Silveira Thaís Gomes Verzignassi
Programa de Pós-Graduação em Ciências da Saúde, Universidade Estadual de Maringá, Maringá, Paraná, Brazil.
Programa de Pós-Graduação em Biociências e Fisiopatologia, Universidade Estadual de Maringá, Maringá, Paraná, Brazil.
Exp Parasitol. 2016 Aug;167:83-93. doi: 10.1016/j.exppara.2016.05.003. Epub 2016 May 13.
Leishmaniasis is a group of diseases that presents various clinical manifestations. Many studies have shown that the parasite plays an important role in the clinical manifestations and prognosis of this disease. The cutaneous and mucosal forms of American tegumentary leishmaniasis (ATL) are associated with Leishmania (Viannia) braziliensis, which exhibits intraspecific genetic polymorphisms and various clinical manifestations. The present study focused on four different L. braziliensis strains that were isolated from patients with distinct Glucantime(®) treatment responses. The isolates were described based on their molecular, biological, and infective characteristics. Growth patterns in culture medium and different grow phases were analyzed, MID-Logarithimic (Mid-LOG), Logarithimic (LOG) and Stationary (STAT) phases. Complement resistance was evaluated using guinea pig serum. Infection to murine peritoneal macrophages, cytokine and nitric oxide were analyzed. Ultrastructural features were determined by transmission electron microscopy, and molecular characteristics were determined based on random amplified polymorphic DNA (RAPD). All of the L. braziliensis isolates showed typical growth and similar complement sensitivity patterns. Markedly lower infectivity indexes were observed for all strains in the LOG phase, with different cytokine profiles. The ultrastructure analysis revealed distinct differences between the MID-LOG, LOG, and STAT phases. The RAPD results showed a divergence between the isolates of the L. braziliensis. The in vitro characterization of L. braziliensis isolates from humans with different treatment responses using various parameters enabled us to observe differences among the isolates. Molecular and in vivo characterizations are currently under study to improve understanding of the parasite-host interaction that can imply in the clinical manifestation differences.
利什曼病是一组具有多种临床表现的疾病。许多研究表明,该寄生虫在这种疾病的临床表现和预后中起着重要作用。美洲皮肤利什曼病(ATL)的皮肤和黏膜形式与巴西利什曼原虫(Leishmania (Viannia) braziliensis)有关,该原虫表现出种内遗传多态性和多种临床表现。本研究聚焦于从对葡糖胺(Glucantime®)治疗反应不同的患者中分离出的四种不同的巴西利什曼原虫菌株。根据其分子、生物学和感染特性对分离株进行了描述。分析了它们在培养基中的生长模式和不同生长阶段,即中对数期(Mid-LOG)、对数期(LOG)和稳定期(STAT)。使用豚鼠血清评估补体抗性。分析了对小鼠腹腔巨噬细胞的感染、细胞因子和一氧化氮。通过透射电子显微镜确定超微结构特征,并基于随机扩增多态性DNA(RAPD)确定分子特征。所有巴西利什曼原虫分离株均显示出典型的生长和相似的补体敏感性模式。在对数期,所有菌株的感染指数均显著降低,且细胞因子谱不同。超微结构分析揭示了中对数期、对数期和稳定期之间的明显差异。RAPD结果显示巴西利什曼原虫分离株之间存在差异。使用各种参数对来自不同治疗反应患者的巴西利什曼原虫分离株进行体外特征分析,使我们能够观察到分离株之间的差异。目前正在进行分子和体内特征分析,以增进对可能导致临床表现差异的寄生虫-宿主相互作用的理解。