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RNA seq analysis of human monocyte subsets in healthy and dengue subjects

ABSTRACT: In this analysis, sorted classical (CM), intermediate (IM) and non-classical (NCM) monocyte subsets from children under steady state (healthy, H) and dengue febrile illness (Dengue, D) were analyzed for their transcriptional profiles using RNA seq. The monocyte subsets were sorted from peripheral blood cells after excluding CD3, CD19, CD20, CD56, CD66b and NKp30 positive cells and then gating on HLADR positive population to identify CM, IM and NCM subsets based on surface expression of CD16 and CD14. The transcriptional profile of the three monocyte subsets was separately compared in healthy children, in dengue febrile children and in dengue versus healthy states. This study highlights hierarchy of gene expression in classical, intermediate and non-classical monocytes in healthy and dengue febrile conditions.

ORGANISM(S): Homo sapiens

PROVIDER: GSE176079 | GEO | 2022/05/16

REPOSITORIES: GEO

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Project description:Viral infections are among the most common causes for fever without an apparent source (FWS) in young children; however, many febrile children are treated with antibiotics despite the absence of bacterial infection. Adenovirus, human herpesvirus 6 (HHV-6) and enterovirus are detected in children with FWS more often than other viral species. Virus and bacteria interact with pattern recognition receptors in circulating blood leukocytes and trigger specific host transcriptional programs that mediate immune response, and unique transcriptional signatures may be ascertained to discriminate between viral and bacterial causes for children with FWS. Microarray analyses were conducted on peripheral blood samples obtained from 51 pediatric patients with confirmed adenovirus, human herpesvirus 6 (HHV-6), enterovirus or bacterial infection. Whole blood transcriptional profiles could clearly distinguish febrile children from healthy controls, and febrile children with viral infections from afebrile children carrying the same virus. Molecular pathways regulating host immune response were the most affected in febrile children with infection. Pattern recognition programs were prominently activated in all febrile children with infection, while differential activation of transcriptional programs was observed among viral species. Interferon signaling pathway was uniquely activated in children with febrile viral infection, while a different set of pathways was uniquely activated in children with bacterial infection. Transcriptional signatures were identified and classified febrile children with viral or bacterial infection with 87% overall accuracy, an improvement from the current clinical practice of deducing from white blood cell (WBC) count status. Similar degree of accuracy was observed when we validated the signature probes on data sets from an independent study with different microarray platforms. The current study confirms the clinical utility of blood transcriptional analysis, suggests the composition of transcriptional signatures which can be used to ascertain the infectious etiology of febrile young children without an apparent source, thus limit the overuse of antibiotics on febrile children presenting with this common clinical complaint. Total RNA samples extracted from whole blood of young children were processed for hybridization onto Illumina Human-HT12 version 4 beadchips, and differential expression of the transcripts was analyzed between sick children with either viral or bacterial infection and healthy children.

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RNA seq analysis of human monocyte subsets in healthy and dengue subjects

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