Transcriptome Profiling Reveals CD73 and Age-Driven Changes in Neutrophil Responses against Streptococcus pneumoniae
Transcriptome Profiling Reveals CD73 and Age-Driven Changes in Neutrophil Responses against Streptococcus pneumoniae
复制标题
DOI：
10.1128/iai.00258-21
复制
发表时间：
2021-11-01
影响因子：
3.1
通讯作者：
Ghanem, Elsa N. Bou
中科院分区：
文献类型：
--
作者：
Bhalla, Manmeet;Heinzinger, Lauren R.;Ghanem, Elsa N. Bou
中性粒细胞是宿主抵抗肺炎链球菌所必需的，但它们的功能随着年龄的增长而下降。我们之前发现CD73，一种抗菌活性所需的酶，在老年小鼠的中性粒细胞（也称为多形核白细胞[PMNs]）中下调。本研究探讨了肺炎链球菌诱导中性粒细胞的转录变化，以确定由CD73控制并随年龄失调的途径。从野生型（WT）小鼠、老年小鼠和CD73敲除小鼠（CD73KO）中分离的纯骨髓源性中性粒细胞被体外模拟肺炎链球菌攻击或感染。采用RNA测序（RNA- seq）鉴定差异表达基因（DEGs）。我们发现感染在宿主中引发了明显的全球转录变化，这些变化在CD73KO中性粒细胞中最强。令人惊讶的是，在所有感染组中，下调的基因多于上调的基因。deg下调表明老年和CD73KO宿主的免疫反应受到抑制。进一步分析显示，CD73KO中性粒细胞表达的长链非编码rna （lncRNAs）数量高于WT对照组。预测的网络分析表明，cd73ko特异性的IncRNAs控制着几种信号通路。我们发现c-Jun n-末端激酶(JNK)-丝裂原活化蛋白激酶（MAPK）通路的基因在CD73KO小鼠和WT老年小鼠感染后上调，而在WT幼年小鼠中没有上调。这与功能差异相对应，因为在感染CD73KO小鼠和老年小鼠的中性粒细胞中，下游AP-1转录因子组分c-Jun的磷酸化水平明显更高。重要的是，抑制JNK/AP-1恢复了这些中性粒细胞杀死肺炎链球菌的能力。总之，我们的研究结果表明，中性粒细胞修改其基因表达以更好地适应细菌感染的能力部分受CD73调节，并随着年龄的增长而下降。
Neutrophils are required for host resistance against Streptococcus pneumoniae, but their function declines with age. We previously found that CD73, an enzyme required for antimicrobial activity, is downregulated in neutrophils (also known as polymorphonuclear leukocytes [PMNs]) from aged mice. This study explored transcriptional changes in neutrophils induced by S. pneumoniae to identify pathways controlled by CD73 and dysregulated with age. Pure bone marrow-derived neutrophils isolated from wild-type (WT) young and old and CD73 knockout (CD73KO) young mice were mock challenged or infected with S. pneumoniae ex vivo. RNA sequencing (RNA-Seq) was performed to identify differentially expressed genes (DEGs). We found that infection triggered distinct global transcriptional changes across hosts that were strongest in CD73KO neutrophils. Surprisingly, there were more downregulated than upregulated genes in all groups upon infection. Downregulated DEGs indicated a dampening of immune responses in old and CD73KO hosts. Further analysis revealed that CD73KO neutrophils expressed higher numbers of long noncoding RNAs (lncRNAs) than those in WT controls. Predicted network analysis indicated that CD73KO-specific IncRNAs control several signaling pathways. We found that genes in the c-Jun N-terminal kinase (JNK)-mitogen-activated protein kinase (MAPK) pathway were upregulated upon infection in CD73KO mice and in WT old mice, but not in WT young mice. This corresponded to functional differences, as phosphorylation of the downstream AP-1 transcription factor component c-Jun was significantly higher in neutrophils from infected CD73KO mice and old mice. Importantly, inhibition of JNK/AP-1 rescued the ability of these neutrophils to kill S. pneumoniae. Together, our findings revealed that the ability of neutrophils to modify their gene expression to better adapt to bacterial infection is in part regulated by CD73 and declines with age.