Garcia Ana F, Benchimol M, Alderete J F
Department Microbiology, UTHSCSA, 7703 Floyd Curl Dr., San Antonio, TX 78229-3900, USA.
Infect Immun. 2005 May;73(5):2602-10. doi: 10.1128/IAI.73.5.2602-2610.2005.
Trichomonas vaginalis secretes putrescine that is readily detected in vaginal secretions. We wanted to examine the effect of decreased putrescine synthesis by inhibition of ornithine decarboxylase (ODC) on T. vaginalis. One reason is because inhibition of Tritrichomonas foetus ODC results in growth arrest, destruction of hydrogenosomes, and decreased amounts of hydrogenosomal enzymes. Treatment of T. vaginalis T016 with >/=20 mM 1,4-diamino-2-butanone (DAB) to inhibit ODC resulted in growth arrest, which was reversed by addition of exogenous putrescine. No similar reversal of growth arrest was achieved with the polyamines spermine or spermidine or with iron. Electron microscopic examination of control versus DAB-treated trichomonads did not reveal any adverse effects on the number and integrity of hydrogenosomes. Further, the adhesins AP65, AP51, and AP33 mediating binding to immortalized vaginal epithelial cells (VECs) share identity to enzymes of the hydrogenosome organelle, and there was no difference in amounts of adhesins between control versus DAB-treated T. vaginalis parasites. Likewise, similar patterns and extent of fluorescence were evident for the prominent AP65 adhesin. Surprisingly, DAB treatment increased by 4- to 20-fold above untreated trichomonads handled identically the level of adherence mediated by adhesins. Interestingly, the enhanced attachment to VECs was reversed by exogenous putrescine added to DAB-treated trichomonads. Equally noteworthy was that DAB-treated T. vaginalis with enhanced adherence did not possess the previously reported ability to kill host cells in a contact-dependent fashion mediated by cysteine proteinases, and total cysteine proteinase activity patterns were identical between control and DAB-treated trichomonads. Overall, these data suggest that polyamine metabolism and secreted putrescine are linked to host cell adherence and cytotoxicity.
阴道毛滴虫分泌的腐胺很容易在阴道分泌物中被检测到。我们想要研究通过抑制鸟氨酸脱羧酶(ODC)来减少腐胺合成对阴道毛滴虫的影响。一个原因是,抑制胎儿三毛滴虫的ODC会导致生长停滞、氢化酶体破坏以及氢化酶体酶数量减少。用≥20 mM的1,4 - 二氨基 - 2 - 丁酮(DAB)处理阴道毛滴虫T016以抑制ODC会导致生长停滞,添加外源性腐胺可使其逆转。用多胺精胺或亚精胺或铁处理未能实现类似的生长停滞逆转。对对照和经DAB处理的滴虫进行电子显微镜检查未发现对氢化酶体的数量和完整性有任何不利影响。此外,介导与永生化阴道上皮细胞(VECs)结合的粘附素AP65、AP51和AP33与氢化酶体细胞器的酶具有同源性,对照和经DAB处理的阴道毛滴虫寄生虫之间的粘附素数量没有差异。同样,突出的AP65粘附素的荧光模式和程度也相似。令人惊讶的是,DAB处理使粘附素介导的粘附水平比未处理的相同处理的滴虫增加了4至20倍。有趣的是,向经DAB处理的滴虫中添加外源性腐胺可逆转对VECs增强的附着。同样值得注意的是,粘附增强的经DAB处理的阴道毛滴虫不具备先前报道的以半胱氨酸蛋白酶介导的接触依赖性方式杀死宿主细胞的能力,对照和经DAB处理的滴虫之间的总半胱氨酸蛋白酶活性模式相同。总体而言,这些数据表明多胺代谢和分泌的腐胺与宿主细胞粘附和细胞毒性有关。