人类诺如病毒(HuNoV)是一组不同的RNA病毒,可引起地方性和大流行性急性病毒性胃肠炎。以前,我们报道了许多HuNoV菌株需要胆汁或胆汁酸(BA)才能感染人空肠肠样培养物。BA对于引起大流行的GII.4HuNoV毒株的复制不是必需的。我们发现疏水性BA糖脱氧胆酸(GCDCA)促进了空肠肠样物质中BA依赖性菌株GII.3的复制。此外,我们发现抑制G蛋白偶联BA受体,鞘氨醇-1-磷酸受体2(S1PR2),通过JTE-013,剂量依赖性地减少了GII.3感染,并抑制了类肠样物质中GII.3细胞的摄取。在这里,我们试图确定其他BA依赖性HuNoV菌株是否需要S1PR2,BA非依赖性GII.4,以及来自其他小肠段的HIE中BA依赖性HuNoV感染是否需要S1PR2。我们发现了第二种S1PR2抑制剂,GLPG2938,剂量依赖性地减少GII.3感染,和S1PR2激动剂(CYM-5520)在不存在GCDCA的情况下增强GII.3复制。在JTE-013和CYM-5520存在下,GII.3复制也被终止。JTE-013在空肠HIE中抑制S1PR2可减少GI.1,GII.3和GII.17(BA依赖性),但不减少GII.4悉尼(BA非依赖性)感染,提供了在HuNoV感染中菌株特异性差异的额外证据。最后,GII.3十二指肠感染,空肠,来自同一个体的回肠系被S1PR2抑制减少,表明小肠多个节段之间BA依赖性感染的共同机制。我们的结果支持一个模型,其中BA依赖性HuNoV利用BA对S1PR2的影响感染整个小肠。IMPORTANCEHumannorovirus(HuNoVs)是重要的病毒性人类病原体,其引起爆发和散发性胃肠炎。这些病毒是多种多样的,许多菌株能够感染人类。我们先前的研究已经确定了疏水性胆汁酸(BA)感染肠上皮细胞的菌株特异性要求。此外,我们鉴定了一种BA受体,鞘氨醇-1-磷酸受体2(S1PR2),BA依赖性菌株感染所需的。为了更好地了解各种HuNoV菌株如何进入和感染小肠以及S1PR2在HuNoV感染中的作用,我们使用扩展的肠道类肠细胞系库评估了其他HuNoV菌株的感染。我们发现多个BA依赖菌株,但不是独立于BA的菌株,所有感染都需要S1PR2。此外,BA依赖性感染需要在小肠的多个区段中的S1PR2。一起,这些结果表明,S1PR2作为BA依赖性HuNoV感染的潜在治疗靶点具有价值.
Human noroviruses (HuNoVs) are a diverse group of RNA viruses that cause endemic and pandemic acute viral gastroenteritis. Previously, we reported that many HuNoV strains require bile or bile acid (BA) to infect human jejunal intestinal enteroid cultures. BA was not essential for the replication of a pandemic-causing GII.4 HuNoV strain. We found the hydrophobic BA glycochenodeoxycholic acid (GCDCA) promotes the replication of the BA-dependent strain GII.3 in jejunal enteroids. Furthermore, we found that inhibition of the G-protein-coupled BA receptor, sphingosine-1-phosphate receptor 2 (S1PR2), by JTE-013, reduced GII.3 infection dose-dependently and inhibited GII.3 cellular uptake in enteroids. Herein, we sought to determine whether S1PR2 is required for other BA-dependent HuNoV strains, the BA-independent GII.4, and whether S1PR2 is required for BA-dependent HuNoV infection in HIEs from other small intestinal segments. We found a second S1PR2 inhibitor, GLPG2938, reduces GII.3 infection dose-dependently, and an S1PR2 agonist (CYM-5520) enhances GII.3 replication in the absence of GCDCA. GII.3 replication also is abrogated in the presence of JTE-013 and CYM-5520. JTE-013 inhibition of S1PR2 in jejunal HIEs reduces GI.1, GII.3, and GII.17 (BA-dependent) but not GII.4 Sydney (BA-independent) infection, providing additional evidence of strain-specific differences in HuNoV infection. Finally, GII.3 infection of duodenal, jejunal, and ileal lines derived from the same individual is reduced with S1PR2 inhibition, indicating a common mechanism of BA-dependent infection among multiple segments of the small intestine. Our results support a model where BA-dependent HuNoVs exploit BA effects on S1PR2 to infect the entire small intestine.IMPORTANCEHuman noroviruses (HuNoVs) are important viral human pathogens that cause both outbreaks and sporadic gastroenteritis. These viruses are diverse, and many strains are capable of infecting humans. Our previous studies have identified strain-specific requirements for hydrophobic bile acids (BAs) to infect intestinal epithelial cells. Moreover, we identified a BA receptor, sphingosine-1-phosphate receptor 2 (S1PR2), required for infection by a BA-dependent strain. To better understand how various HuNoV strains enter and infect the small intestine and the role of S1PR2 in HuNoV infection, we evaluated infection by additional HuNoV strains using an expanded repertoire of intestinal enteroid cell lines. We found that multiple BA-dependent strains, but not a BA-independent strain, all require S1PR2 for infection. In addition, BA-dependent infection requires S1PR2 in multiple segments of the small intestine. Together, these results indicate that S1PR2 has value as a potential therapeutic target for BA-dependent HuNoV infection.