Gaspar Emanuelle Baldo, Mortara Renato Arruda, Andrade Luciana O, da Silva Claudio Vieira
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, Rua Botucatu 862, 6 Degrees Andar, São Paulo 04023-062 SP, Brazil.
Biochem Biophys Res Commun. 2009 Jun 26;384(2):265-9. doi: 10.1016/j.bbrc.2009.04.114. Epub 2009 May 4.
Trypanosoma cruzi is an obligate intracellular organism in vertebrate hosts. Lysosomes are involved in parasite invasion. LAMP-1 and LAMP-2 are the most abundant glycoproteins of the lysosomal membrane. This study is the first report on the invasion of T. cruzi extracellular amastigotes (EA) in single LAMP-1 or LAMP-2 knockouts, respectively, or in two independent LAMP-1/2 double-knockout cell lines. When compared to their respective wild type clones, the EA show higher infectivity in LAMP-2 knockouts, but no difference was seen in LAMP-1 knockout cells. Similarly, EA invasion rate was higher for one of the double knockout clones but not for the other. Higher lysosomal exocytosis correlated with a higher invasion rate and early lysosomal marker acquisition. These findings suggest that lysosomal exocytosis is important to EA cell invasion. Also, phagolysosome maturation in knockout cell lines differed from previous results revealing that EA enter cells by a mechanism other than receptor-mediated phagocytosis.
克氏锥虫是脊椎动物宿主中的专性细胞内寄生虫。溶酶体参与寄生虫的入侵。LAMP - 1和LAMP - 2是溶酶体膜中含量最丰富的糖蛋白。本研究首次报道了克氏锥虫细胞外无鞭毛体(EA)分别侵入单个LAMP - 1或LAMP - 2基因敲除细胞,以及两种独立的LAMP - 1/2双基因敲除细胞系的情况。与各自的野生型克隆相比,EA在LAMP - 2基因敲除细胞中的感染性更高,但在LAMP - 1基因敲除细胞中未观察到差异。同样,其中一个双基因敲除克隆的EA入侵率较高,而另一个则不然。较高的溶酶体胞吐作用与较高的入侵率和早期溶酶体标志物的获得相关。这些发现表明溶酶体胞吐作用对EA细胞入侵很重要。此外,基因敲除细胞系中的吞噬溶酶体成熟与先前的结果不同,这表明EA通过受体介导的吞噬作用以外的机制进入细胞。