Pereira-Neves Antonio, Benchimol Marlene
Laboratório de Ultraestrutura Celular, Universidade Santa Ursula, Rua Jornalista Orlando Dantas 59, Botafogo, CEP 22231-010, Rio de Janeiro, Brazil.
Biol Cell. 2007 Feb;99(2):87-101. doi: 10.1042/BC20060084.
The parasitic protozoan Trichomonas vaginalis is the causative agent of trichomoniasis, a sexually transmitted disease. The phagocytic activity of this parasite has not been completely elucidated. In order to better understand the mechanisms of trichomonal phagocytosis, we have studied the in vitro capacity of T. vaginalis to phagocytose and degrade Saccharomyces cerevisiae cells.
To analyse the phagocytic ability and capacity, two isolates of T. vaginalis presenting different virulence grades were used. Complementary techniques, such as fluorescence microscopy, computer-based fluorescence analysis, scanning and transmission electron microscopy and the use of drugs that interfere with the actin microfilaments, were used in order to follow the behaviour of the actin cytoskeleton during phagocytosis of yeast cells by T. vaginalis. It was concluded that: (1) T. vaginalis changes its shape rapidly and engulfs the yeast cells, which are almost as large as the parasite; (2) long-term and fresh cultures are able to phagocytose, although the low-virulence strain JT demonstrated a lower activity when compared with the highly virulent T016 isolate; (3) the T016 strain exhibited an amoeboid morphology during the internalization of yeast cells in contrast with the JT strain; (4) attachment of yeast cells to the parasite occurs via the whole cell surface, including both anterior and recurrent flagella; (5) two forms of phagocytosis were observed: a 'sinking' process without any apparent participation of plasma membrane extensions and the classical phagocytosis where pseudopodia are extended toward the target cell; (6) the internalized S. cerevisiae are digested in lysosomes; (7) competitor sugars D-mannose or L-fucose inhibit the phagocytosis, and inhibition was 1.67 times higher in long-term cultured JT than that of the parasites from fresh isolate T016; (8) a thick layer of actin microfilaments was present underlying the plasma membrane, and especially in the pseudopodia and around the phagocytosed particles; (9) a dramatic change in the distribution pattern of fibrillar actin occurred during phagocytosis; (10) cytochalasin D depressed the phagocytosis; (11) a non-specific recognition and phagocytosis of yeast cells by T. vaginalis is mediated by a mannose receptor present on the parasite surface; (12) the phagocytic process may occur simultaneously during mitosis of the parasite.
寄生原生动物阴道毛滴虫是性传播疾病滴虫病的病原体。这种寄生虫的吞噬活性尚未完全阐明。为了更好地理解滴虫吞噬作用的机制,我们研究了阴道毛滴虫在体外吞噬和降解酿酒酵母细胞的能力。
为了分析吞噬能力,使用了两种具有不同毒力等级的阴道毛滴虫分离株。采用了荧光显微镜、基于计算机的荧光分析、扫描和透射电子显微镜以及使用干扰肌动蛋白微丝的药物等补充技术,以追踪阴道毛滴虫吞噬酵母细胞过程中肌动蛋白细胞骨架的行为。得出以下结论:(1)阴道毛滴虫迅速改变形状并吞噬几乎与寄生虫大小相同的酵母细胞;(2)长期培养和新鲜培养的菌株都能够进行吞噬,尽管低毒力菌株JT与高毒力T016分离株相比活性较低;(3)与JT菌株相比,T016菌株在酵母细胞内化过程中呈现出阿米巴样形态;(4)酵母细胞通过整个细胞表面附着于寄生虫,包括前鞭毛和后鞭毛;(5)观察到两种吞噬形式:一种是没有明显质膜延伸参与的“下沉”过程,以及伪足向靶细胞延伸的经典吞噬作用;(6)内化的酿酒酵母在溶酶体中被消化;(7)竞争性糖类D-甘露糖或L-岩藻糖抑制吞噬作用,长期培养的JT菌株的抑制作用比新鲜分离的T016菌株的寄生虫高1.67倍;(8)质膜下方存在一层厚厚的肌动蛋白微丝,特别是在伪足和吞噬颗粒周围;(9)吞噬过程中丝状肌动蛋白的分布模式发生了显著变化;(10)细胞松弛素D抑制吞噬作用;(11)阴道毛滴虫对酵母细胞的非特异性识别和吞噬作用由寄生虫表面存在的甘露糖受体介导;(12)吞噬过程可能在寄生虫有丝分裂期间同时发生。