Immune-related gene expression in severe periodontitis assessed by NanoString technology: A preliminary study

Abstract

Periodontitis is an inflammatory disease characterized by the destruction of the periodontal attachment apparatus, which includes alveolar bone, periodontal ligament, and cementum. This destruction is driven by a dysregulated host immune response to pathogenic subgingival biofilm. The present preliminary study aimed to evaluate immune-related gene expression patterns in patients with stage III/IV periodontitis utilizing the NanoString nCounter® platform. Unstimulated saliva samples were collected from twelve individuals: ten with severe periodontitis (stage III/IV) and two periodontally healthy controls. Total RNA was isolated and analyzed using the nCounter® Human Inflammation Panel, which profiles 249 inflammation-associated human genes. Data normalization and differential expression analysis were performed with nSolver™ software. Following quality control, genes with low expression (mean normalized counts < 20) were excluded, resulting in 89 genes available for comparison. Among these, 26 genes (29.2%) met a predefined effect-size threshold (|log₂FC| ≥ 1), comprising 23 upregulated and 3 downregulated transcripts in the periodontitis group. Notably, the upregulated genes HLA-DRB1 (p = 0.003; FDR = 0.267) and CCR1 (p = 0.007; FDR = 0.312) exhibited relatively large log₂ fold changes and the lowest unadjusted p-values; however, neither retained significance after FDR correction. These findings underscore the feasibility of salivary gene expression profiling as a method for identifying molecular markers associated with disease severity. Given their roles in immune activation and leukocyte recruitment, HLA-DRB1 and CCR1 emerge as potential biomarker candidates for detection, risk stratification, and therapeutic monitoring in periodontitis, necessitating validation in larger, well-characterized cohorts.