e529ad12283f1f0a4acecc92066506f6 283215.pdf 657dc26355afb1507df138d49648fb9e155b8129 283215.pdf d2ea658df1a2ded98ea49a893531cfe51c8ecce97abf0fe54eae18b35e7917fb 283215.pdf Title: Increase of Circulating Monocyte–Platelet Conjugates in Rheumatoid Arthritis Responders to IL-6 Blockage Subject: Platelets (PLT) bind to a significant percentage of circulating monocytes and this immunomodulatory interaction is increased in several inflammatory and autoimmune conditions. The therapeutic blockage of IL-6 with Tocilizumab (TCZ) alters PLT and the phenotype and function of monocytes in rheumatoid arthritis (RA). However, the relationship between monocyte–PLT conjugates (CD14+PLT+) and clinical and immunological variables and the regulation of this interaction by IL-6 blockage are still unknown. Here, we compared the presence of monocyte–PLT conjugates (CD14+PLT+) and membrane CD162 expression using flow cytometry, and, by ELISA, the markers of PLT activation (sCD62P and sCD40L) in healthy donors (HD) and patients with long-standing RA before TCZ (baseline). We found higher percentages and absolute counts of CD14+PLT+, and higher plasmatic levels of sCD62P and sCD40L but lower CD162 expression on monocytes from RA patients than those from HD. Additionally, the levels of CD14+PLT+ inversely correlated with inflammatory parameters. Interestingly, 95% of patients with lower percentages of CD14+PLT+ and only 63% of patients with higher percentages of CD14+PLT+ achieved a EULAR-defined response at four weeks (p = 0.036). After TCZ, the percentage of CD14+PLT+ increased in 92% of RA patients who achieved 12 w-remission (p < 0.001). Our results suggest that the binding of PLTs has a modulatory effect, accentuated by the increased binding of PLTs to monocytes in response to the therapeutic blockage of IL-6. Keywords: platelets; monocytes; rheumatoid arthritis; tocilizumab; immune modulation; inflammation; immunity Author: Anaís Mariscal, Carlos Zamora, César Díaz-Torné, Mᵃ Àngels Ortiz, Juan José de Agustín, Delia Reina, Paula Estrada, Patricia Moya, Héctor Corominas and Sílvia Vidal Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Sat May 21 05:34:25 2022 CEST ModDate: Sat May 21 05:37:35 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 13 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1619125 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- WBZAFJ+VnURWPalladioL Type 1 Custom yes yes no 10 0 CKUPOS+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 VTJFSN+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 YKHVLG+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 MDJLZU+EURM10 Type 1 Builtin 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edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Children: Item: Title: Circulating CD14+PLT+ in RA Patients Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Relationship between CD14+PLT+ and mCD162 Expression on Monocytes with Clinical RA Parameters Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Relationship between CD14+PLT+ and mCD162 Expression on Monocytes with Clinical Outcome in RA Patients after TCZ Treatment Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: IL-6 and TCZ Effects over PLT Binding to Monocytes, mCD162 Expression on Monocytes and PLT Activation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Children: Item: Title: Patients Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Item: Title: Peripheral Blood Staining and Flow Cytometry Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Item: Title: Plasma Analysis of RF, ACPAs, and Cytokines Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@e720b71 Item: Title: Peripheral Blood Cultures Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1b26f7b2 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@491cc5c9 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@74ad1f1f Info: Title: Increase of Circulating Monocyte Platelet Conjugates in Rheumatoid Arthritis Responders to IL-6 Blockage Author: ぁのちロび〠きちひどびっちぬ〬〠ぃちひぬはび〠ずちねはひち〬〠ぃラびちひ〠いロちぺ〭ごはひのラ〬〠き㕃〠ダのでづぬび〠くひぴどぺ〬〠おふちの〠おはびラ〠つづ〠ぁでふびぴロの〬〠いづぬどち〠げづどのち〬〠ぐちふぬち〠ぅびぴひちつち〬〠ぐちぴひどっどち〠きはべち〬〠えラっぴはひ〠ぃはひはねどのちび〠ちのつ〠こロぬぶどち〠ざどつちぬ Subject: Platelets (PLT) bind to a significant percentage of circulating monocytes and this immunomodulatory interaction is increased in several inflammatory and autoimmune conditions. The therapeutic blockage of IL-6 with Tocilizumab (TCZ) alters PLT and the phenotype and function of monocytes in rheumatoid arthritis (RA). However, the relationship between monocyte PLT conjugates (CD14+PLT+) and clinical and immunological variables and the regulation of this interaction by IL-6 blockage are still unknown. Here, we compared the presence of monocyte PLT conjugates (CD14+PLT+) and membrane CD162 expression using flow cytometry, and, by ELISA, the markers of PLT activation (sCD62P and sCD40L) in healthy donors (HD) and patients with long-standing RA before TCZ (baseline). We found higher percentages and absolute counts of CD14+PLT+, and higher plasmatic levels of sCD62P and sCD40L but lower CD162 expression on monocytes from RA patients than those from HD. Additionally, the levels of CD14+PLT+ inversely correlated with inflammatory parameters. Interestingly, 95% of patients with lower percentages of CD14+PLT+ and only 63% of patients with higher percentages of CD14+PLT+ achieved a EULAR-defined response at four weeks (p = 0.036). After TCZ, the percentage of CD14+PLT+ increased in 92% of RA patients who achieved 12 w-remission (p < 0.001). Our results suggest that the binding of PLTs has a modulatory effect, accentuated by the increased binding of PLTs to monocytes in response to the therapeutic blockage of IL-6. 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