A Mr 34,000 proinflammatory outer membrane protein (oipA) of Helicobacter pylori
A Mr 34,000 proinflammatory outer membrane protein (oipA) of Helicobacter pylori
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DOI：
10.1073/pnas.130079797
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发表时间：
2000-06-20
影响因子：
11.1
通讯作者：
Graham, DY
中科院分区：
文献类型：
--
作者：
Yamaoka, Y;Kwon, DH;Graham, DY
幽门螺杆菌的全基因组序列揭示了一个由32个外膜蛋白（OMP）组成的家族。我们检测了四种OMP（HP 0638、HP 0796、HP 1501和babA 2）对促炎细胞因子IL-8产生的影响。这四种外膜蛋白的突变体以及来自H.来自美国和日本的pylori是通过将抗氯霉素盒插入基因中构建的。22对亲本和突变体H.幽门螺杆菌菌株，以及160个临床分离株（80个来自日本，80个来自美国），与胃癌细胞系共培养。ELISA法检测细胞培养上清中IL-8的产生和细胞粘附。HP 0796、HP 1501、babA 2和galE基因敲除对IL-8的产生没有显着影响。在81%的cag阳性菌株中，HP 0638基因的敲除使IL-8的产生减少约50%。其中IL-8水平通过HP 0638敲除未改变的三个cag阳性菌株在5'区域具有五个或七个CT二核苷酸重复，导致移码和截短。具有天然失活的HP 0638基因的菌株均来自美国;日本的病毒株总是处于“开启”状态，因此，平均而言，可能更具毒性。尽管cag阴性分离株产生有限的IL-8应答，但含有功能性HP 0638基因的cag阴性菌株产生的IL-8比cag阴性非功能性HP 0638菌株高3倍以上。我们推测，功能性HP 0638基因可能是一个重要的毒力因子，与H.幽门感染我们将HP 0638基因命名为外炎性蛋白（oipA）。
The complete genome sequence revealed a family of 32 outer membrane proteins (OMPs) in Helicobacter pylori. We examined the effect of four OMPs (HP0638, HP0796, HP1501, and babA2) on the production of the proinflammatory cytokine, IL-8. Mutants of the four OMPs, as well as cagE and galE from H. pylori from the U.S. and Japan, were constructed by inserting a chloramphenicol-resistant cassette into the gene. Twenty-two pairs of parental and mutant H. pylori strains, as well as 160 clinical isolates (80 from Japanese and 80 from U.S.), were cocultured with gastric cancer cell lines. IL-8 production in the supernatant and adhesion was assayed by ELISA. HP0796, HP1501, babA2, and galE gene knockouts had no significant effect on IL-8 production. Knockout of the HP0638 gene in 81% of cag-positive strains reduced IL-8 production approximately 50%. The three cag-positive strains in which IL-8 levels were unchanged by HP0638 knockout had five or seven CT dinucleotide repeats in the 5' region, resulting in a frame shift and truncation. Strains with naturally inactive HP0638 gene were all from the U.S.; Japanese strains were always "on" and thus, on average, may be more virulent. Although cag-negative isolates produced a limited IL-8 response, cag-negative strains that contained a functional HP0638 gene produced more than 3-fold greater IL-8 than cag-negative nonfunctional HP0638 strains. We hypothesize that functional HP0638 gene may be an important virulence factor in relation to the risk of clinically significant outcomes of H. pylori infection. We denote HP0638 gene as outer inflammatory protein (oipA).