Comparison of differential gene expression in blood PBMCs from healthy controls, patients with rheumatoid arthritis,and patients with lupus using single-cell transcriptome sequencing analysis.

Single-cell transcriptome sequencing was used to detect and compare differentially expressed genes in PBMCs from healthy controls in patients with rheumatoid arthritis, systemic lupus erythematosus patients. Overall design: The study included three healthy controls (NC group), three patients with rheumatoid arthritis, three newly diagnosed and untreated patients with systemic lupus erythematosus (SLE group). Inclusion criteria for rheumatoid arthritis patients required seropositivity for antibodies to cyclic citrullinated peptide (CCP) and strict adherence to the American College of Rheumatology (ACR) 2010 classification criteria for rheumatoid arthritis, with the absence of any other rheumatic diseases. Patients diagnosed with systemic lupus erythematosus (SLE) were classified according to the 2019 European League Against Rheumatism (EULAR) and American College of Rheumatology (ACR) EULAR/ACR criteria. Patients with concurrent immunological conditions such as malignancy or rheumatoid arthritis were excluded from the study. Healthy controls consisted of individuals undergoing medical examinations during the same timeframe. Approval for this study was obtained from the Medical Ethics Committee of the First Affiliated Hospital of Shandong Second Medical University. Prior to testing, all participants provided written informed consent and the studies adhered to the ethical guidelines outlined in the Declaration of Helsinki. Peripheral blood samples were collected from patients, and peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-Paque gradient centrifugation. The freshly isolated PBMCs were resuspended in fetal bovine serum with 10% dimethyl sulfoxide, frozen at -80°C for at least 4 hours, stored in liquid nitrogen, and subsequently utilized for single-cell transcriptome sequencing (scRNA-seq).