Evans D J, Frank D W, Finck-Barbançon V, Wu C, Fleiszig S M
School of Optometry, University of California, Berkeley, 94720, USA.
Infect Immun. 1998 Apr;66(4):1453-9. doi: 10.1128/IAI.66.4.1453-1459.1998.
Pseudomonas aeruginosa clinical isolates exhibit invasive or cytotoxic phenotypes. Cytotoxic strains acquire some of the characteristics of invasive strains when a regulatory gene, exsA, that controls the expression of several extracellular proteins, is inactivated. exsA mutants are not cytotoxic and can be detected within epithelial cells by gentamicin survival assays. The purpose of this study was to determine whether epithelial cell invasion precedes and/or is essential for cytotoxicity. This was tested by measuring invasion (gentamicin survival) and cytotoxicity (trypan blue staining) of PA103 mutants deficient in specific exsA-regulated proteins and by testing the effect of drugs that inhibit invasion for their effect on cytotoxicity. A transposon mutant in the exsA-regulated extracellular factor exoU was neither cytotoxic nor invasive. Furthermore, several of the drugs that inhibited invasion did not prevent cytotoxicity. These results show that invasion and cytotoxicity are mutually exclusive events, inversely regulated by an exsA-encoded invasion inhibitor(s). Both involve host cell protein tyrosine kinase (PTK) activity, but they differ in that invasion requires Src family tyrosine kinases and calcium-calmodulin activity. PTK inhibitor drugs such as genistein may have therapeutic potential through their ability to block both invasive and cytotoxicity pathways via an action on the host cell.
铜绿假单胞菌临床分离株表现出侵袭性或细胞毒性表型。当控制几种细胞外蛋白表达的调控基因exsA失活时,细胞毒性菌株获得了一些侵袭性菌株的特征。exsA突变体没有细胞毒性,可通过庆大霉素存活试验在上皮细胞内检测到。本研究的目的是确定上皮细胞侵袭是否先于细胞毒性和/或对细胞毒性是否必不可少。通过测量缺乏特定exsA调控蛋白的PA103突变体的侵袭(庆大霉素存活)和细胞毒性(台盼蓝染色),以及测试抑制侵袭的药物对细胞毒性的影响来进行验证。exsA调控的细胞外因子exoU中的转座子突变体既没有细胞毒性也没有侵袭性。此外,几种抑制侵袭的药物并不能阻止细胞毒性。这些结果表明,侵袭和细胞毒性是相互排斥的事件,由exsA编码的侵袭抑制剂反向调节。两者都涉及宿主细胞蛋白酪氨酸激酶(PTK)活性,但不同之处在于侵袭需要Src家族酪氨酸激酶和钙调蛋白活性。诸如染料木黄酮等PTK抑制剂药物可能具有治疗潜力,因为它们能够通过作用于宿主细胞来阻断侵袭和细胞毒性途径。