413dd71c275833ef8f88ab0ad0d29cf9 pmc_10813161.pdf 2053ee6e25dd7c701dff8722e65215bedb08c5d7 pmc_10813161.pdf 503d5bfd984e2596435d64811f739cccaa3ae7f77411206ba94a36d5113a77c2 pmc_10813161.pdf Title: Altered CD39 and CD73 Expression in Rheumatoid Arthritis: Implications for Disease Activity and Treatment Response Subject: In rheumatoid arthritis (RA) synovium, ATP, and ADP are released, sparking inflammation. Ectoenzymes CD39 and CD73 metabolize these purine nucleotides, generating anti-inflammatory adenosine. Therefore, dysregulated CD39 and CD73 expression may impact RA development. We assessed CD39 and CD73 expression in peripheral blood from 15 healthy controls (Cs) and 35 RA patients at baseline and after 3 and 6 months of tocilizumab treatment using flow cytometry. Additionally, ectoenzyme expression was examined on cultured T cells to understand activation and IL-6 effects. At baseline, RA patients exhibited a lower CD8+CD39-CD73+ cell percentage, which inversely correlated with DAS28. Additionally, they had lower percentages of Treg CD39+CD73+ and CD39-CD73- cells. Good responders tended to have lower B CD39+CD73+ cell percentages at baseline and 3 months. Additionally, Treg, CD8+ T and B cells inversely correlated with DAS28. T-cell activation increased CD39 and decreased CD73 expression, regardless of IL-6. IL-6 reduced IFN-secreting CD4+ T-cell percentage in Cs, but increased the percentage of IFN-secreting CD4+ and CD8+ T cells in RA patients. These findings indicate differing CD39 and CD73 expression in RA and Cs, influenced by T-cell activation and IL-6. Correlations between these molecules and RA activity suggest their role in dysregulated inflammation in RA. Keywords: rheumatoid arthritis; CD39; CD73; tocilizumab; IL-6 Author: María Angels Ortiz, Cesar Diaz-Torné, Juan Jose De Agustin, Paula Estrada, Delia Reina, María Victoria Hernandez, Hye Sang, Carlos Zamora, Elisabet Cantó, Hector Corominas and Silvia Vidal Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Wed Dec 20 02:20:31 2023 CET ModDate: Wed Dec 20 02:28:28 2023 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 17 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3260164 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- XWOCXD+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 HKBJPQ+VnURWPalladioL Type 1 Custom yes yes yes 16 0 WZEBNH+EURM10 Type 1 Builtin yes yes yes 21 0 GNNEVO+URWPalladioL-Bold Type 1 Custom yes yes yes 26 0 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PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-04-24 02:37:58 CEST Size: 3260164 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 559 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Patient Samples and Study Design Destination: subsection.2.1 Item: Title: Serum Analysis of RF and ACPAs Destination: subsection.2.2 Item: Title: Flow Cytometry Destination: subsection.2.3 Item: Title: Luminescence Assays for Quantification of Plasmatic Levels of ATP and ADP Destination: subsection.2.4 Item: Title: Cellular ATP Consumption Assay Destination: subsection.2.5 Item: Title: Cytokine Levels in Serum and Supernatants Destination: subsection.2.6 Item: Title: Cell Culture Preparation Destination: subsection.2.7 Item: Title: Determination of IFN Secretion Destination: subsection.2.8 Item: Title: Statistical Analysis Destination: subsection.2.9 Item: Title: Results Destination: section.3 Children: Item: Title: Expression of CD39 and CD73 on Peripheral Blood Lymphocytes and Monocytes Destination: subsection.3.1 Item: Title: Relationship between CD39 and CD73 Expression with the Disease Activity Indexes Destination: subsection.3.2 Item: Title: Expression of CD39 and CD73 in Good Responders and Non-Good Responders to TCZ Treatment Destination: subsection.3.3 Item: Title: Plasma ATP Levels in RA Patients at Baseline and during TCZ Treatment Destination: subsection.3.4 Item: Title: The In Vitro Effects of IL-6 on CD39 and CD73 Expression in T Cells Destination: subsection.3.5 Item: Title: The In Vitro Role of IL-6 in Influencing the Capacity of Effector T Cells to Produce IFN Destination: subsection.3.6 Item: Title: Discussion Destination: section.4 Item: Title: References Destination: appendix.A. Info: Title: Altered CD39 and CD73 Expression in Rheumatoid Arthritis: Implications for Disease Activity and Treatment Response Author: María Angels Ortiz, Cesar Diaz-Torné, Juan Jose De Agustin, Paula Estrada, Delia Reina, María Victoria Hernandez, Hye Sang, Carlos Zamora, Elisabet Cantó, Hector Corominas and Silvia Vidal Subject: In rheumatoid arthritis (RA) synovium, ATP, and ADP are released, sparking inflammation. Ectoenzymes CD39 and CD73 metabolize these purine nucleotides, generating anti-inflammatory adenosine. Therefore, dysregulated CD39 and CD73 expression may impact RA development. We assessed CD39 and CD73 expression in peripheral blood from 15 healthy controls (Cs) and 35 RA patients at baseline and after 3 and 6 months of tocilizumab treatment using flow cytometry. Additionally, ectoenzyme expression was examined on cultured T cells to understand activation and IL-6 effects. At baseline, RA patients exhibited a lower CD8+CD39-CD73+ cell percentage, which inversely correlated with DAS28. Additionally, they had lower percentages of Treg CD39+CD73+ and CD39-CD73- cells. Good responders tended to have lower B CD39+CD73+ cell percentages at baseline and 3 months. Additionally, Treg, CD8+ T and B cells inversely correlated with DAS28. T-cell activation increased CD39 and decreased CD73 expression, regardless of IL-6. IL-6 reduced IFN-secreting CD4+ T-cell percentage in Cs, but increased the percentage of IFN-secreting CD4+ and CD8+ T cells in RA patients. These findings indicate differing CD39 and CD73 expression in RA and Cs, influenced by T-cell activation and IL-6. Correlations between these molecules and RA activity suggest their role in dysregulated inflammation in RA. 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