Matos Denise C S, Faccioli Lanuza A P, Cysne-Finkelstein Léa, Luca Paula Mello De, Corte-Real Suzana, Armôa Geraldo R G, Lemes Elezer Monte Blanco, Decote-Ricardo Débora, Mendonça Sergio C F
Instituto de Tecnologia em Imunobiológicos, Fiocruz, Rio de Janeiro, RJ, Brasil.
Mem Inst Oswaldo Cruz. 2010 May;105(3):341-7. doi: 10.1590/s0074-02762010000300018.
Kinetoplastid membrane protein-11 (KMP-11), a protein present in all kinetoplastid protozoa, is considered a potential candidate for a leishmaniasis vaccine. A suitable leishmaniasis vaccine candidate molecule must be expressed in amastigotes, the infective stage for mammals. However, the expression of KMP-11 in Leishmania amastigotes has been a subject of controversy. We evaluated the expression of this molecule in logarithmic and stationary growth phase promastigotes, as well as in amastigotes, of Leishmania amazonensis by immunoblotting, flow cytometry and immunocytochemistry, using a monoclonal antibody against KMP-11. We found that KMP-11 is present in promastigotes and amastigotes. In both stages, the protein was found in association with membrane structures (at the cell surface, flagellar pocket and intracellular vesicles). More importantly, its surface expression is higher in amastigotes than in promastigotes and increases during metacyclogenesis. The increased expression of KMP-11 in metacyclic promastigotes, and especially in amastigotes, indicates a role for this molecule in the parasite relationship with the mammalian host. The presence of this molecule in amastigotes is consistent with the previously demonstrated immunoprotective capacity of vaccine prototypes based on the KMP-11-coding gene and the presence of humoral and cellular immune responses to KMP-11 in Leishmania-infected humans and animals.
动质体膜蛋白11(KMP - 11)是一种存在于所有动质体原生动物中的蛋白质,被认为是利什曼病疫苗的潜在候选物。合适的利什曼病疫苗候选分子必须在无鞭毛体中表达,而无鞭毛体是哺乳动物的感染阶段。然而,KMP - 11在亚马逊利什曼原虫无鞭毛体中的表达一直存在争议。我们使用抗KMP - 11单克隆抗体,通过免疫印迹、流式细胞术和免疫细胞化学方法,评估了该分子在亚马逊利什曼原虫对数生长期和稳定期前鞭毛体以及无鞭毛体中的表达情况。我们发现KMP - 11存在于前鞭毛体和无鞭毛体中。在这两个阶段,该蛋白质都与膜结构相关(位于细胞表面、鞭毛袋和细胞内囊泡)。更重要的是,其表面表达在无鞭毛体中高于前鞭毛体,并且在循环前期发育过程中增加。KMP - 11在循环前期前鞭毛体,尤其是无鞭毛体中的表达增加,表明该分子在寄生虫与哺乳动物宿主的关系中发挥作用。无鞭毛体中该分子的存在与先前基于KMP - 11编码基因的疫苗原型所显示的免疫保护能力以及利什曼原虫感染的人类和动物中对KMP - 11的体液和细胞免疫反应的存在是一致的。