Constitutively Opa-Expressing and Opa-Deficient Neisseria gonorrhoeae Strains Differentially Stimulate and Survive Exposure to Human Neutrophils
Constitutively Opa-Expressing and Opa-Deficient Neisseria gonorrhoeae Strains Differentially Stimulate and Survive Exposure to Human Neutrophils
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DOI：
10.1128/jb.00171-13
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发表时间：
2013-07-01
影响因子：
3.2
通讯作者：
Criss, Alison K.
中科院分区：
文献类型：
--
作者：
Ball, Louise M.;Criss, Alison K.
淋病奈瑟菌（淋球菌[Gc]）不透明相关（Opa）蛋白介导细菌结合和人上皮细胞和中性粒细胞（多形核白细胞[PMN]）的内化。研究Opa蛋白对淋球菌发病机制的贡献是复杂的opa基因的高频相位变化。因此，我们设计了Gc菌株FA 1090的衍生物，其中所有opa基因在读码框内缺失，称为Opaless。无乳蛋白Gc保持一致的Opa阴性（Opa（-）），而主要Opa（-）亲本Gc和缺乏opa基因的“半透明”子集（Δ opaBEGK）的中间体的培养物随机产生Opa阳性（Opa（+））细菌菌落。Opa表达的丧失不影响Gc生长。Opaless Gc在暴露于原代人PMN后存活，并抑制PMN氧化爆发，类似于亲本Opa（-）细菌。值得注意的是，未经调理的蛋白质Gc内化粘附，趋化因子引发，原代人中性粒细胞，通过肌动蛋白依赖性过程。当FA 1090 opaD的非时相可变、符合读框的等位基因被重新引入Opaless Gc中时，细菌诱导了PMN氧化爆发，与Opa（-）细菌相比，OpaD（+）Gc在暴露于PMN后的存活率较低。这些衍生物为评估Opa蛋白在GC生物学中的作用提供了一个强大的系统。
The Neisseria gonorrhoeae (the gonococcus [Gc]) opacity-associated (Opa) proteins mediate bacterial binding and internalization by human epithelial cells and neutrophils (polymorphonuclear leukocytes [PMNs]). Investigating the contribution of Opa proteins to gonococcal pathogenesis is complicated by high-frequency phase variation of the opa genes. We therefore engineered a derivative of Gc strain FA1090 in which all opa genes were deleted in frame, termed Opaless. Opaless Gc remained uniformly Opa negative (Opa(-)), whereas cultures of predominantly Opa(-) parental Gc and an intermediate lacking the "translucent" subset of opa genes (Delta opaBEGK) stochastically gave rise to Opa-positive (Opa(+)) bacterial colonies. Loss of Opa expression did not affect Gc growth. Opaless Gc survived exposure to primary human PMNs and suppressed the PMN oxidative burst akin to parental, Opa(-) bacteria. Notably, unopsonized Opaless Gc was internalized by adherent, chemokine-primed, primary human PMNs, by an actin-dependent process. When a non-phase-variable, in-frame allele of FA1090 opaD was reintroduced into Opaless Gc, the bacteria induced the PMN oxidative burst, and OpaD(+) Gc survived less well after exposure to PMNs compared to Opa(-) bacteria. These derivatives provide a robust system for assessing the role of Opa proteins in Gc biology.