5cc248fda2fa2e0307aa6e1009b51171 pmc_33498698.pdf 37031d1201c7874d92beb3b6f8a2b213598a1728 pmc_33498698.pdf 1dc33e71ab3b0f1ab0c54bf1b2b8a03c73fe810639ca088a45f7c0048ea9a9b4 pmc_33498698.pdf Title: Activated Lymphocytes and Increased Risk of Dermatologic Adverse Events during Sorafenib Therapy for Hepatocellular Carcinoma Subject: Advanced hepatocellular carcinoma patients treated with sorafenib who develop early dermatologic adverse events (eDAEs) have a better prognosis. This may be linked to immune mechanisms, and thus, it is relevant to assess the association between peripheral immunity and the probability of developing eDAEs. Peripheral blood mononuclear cells of 52 HCC patients treated with sorafenib were analyzed at baseline and throughout the first eight weeks of therapy. T, B, Natural Killer cells, and their immune checkpoints expression data were characterized by flow cytometry. Cytokine release and immune-suppression assays were carried out ex vivo. Cox baseline and time-dependent regression models were applied to evaluate the probability of increased risk of eDAEs. DNAM-1, PD-1, CD69, and LAG-3 in T cells, plus CD16 and LAG-3 in NK cells, are significantly associated with the probability of developing eDAEs. While NK DNAM-1+ cells express activation markers, T DNAM-1+ cells induce immune suppression and show immune exhaustion. This is the first study to report an association between immune checkpoints expression in circulating immune cells and the increased incidence of eDAEs. Our results support the hypothesis for an off-target role of sorafenib in immune modulation. We also describe a novel association between DNAM-1 and immune exhaustion in T cells. Keywords: hepatocellular carcinoma; sorafenib; early dermatologic adverse events; peripheral blood mononuclear cells; immune cell phenotyping; PD-1; DNAM-1; CD96 Author: Josep Corominas, Victor Sapena, Marco Sanduzzi-Zamparelli, Cristina Millán, Esther Samper, Neus Llarch, Gemma Iserte, Ferràn Torres, Leonardo G. Da Fonseca, Sergio Muñoz-Martínez, Alejandro Forner, Jordi Bruix, Loreto Boix and María Reig Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Sat Jan 23 11:13:14 2021 CET ModDate: Sat Jan 23 11:16:45 2021 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 20 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2302784 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- HKBJPQ+VnURWPalladioL Type 1 Custom yes yes no 10 0 AYIFBV+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 HIPKHS+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 QWSYMG+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 OTZEDH+VnURWPalladioL-Bold Type 1 Custom yes yes no 28 0 LCJEVJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 37 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 59 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 63 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 67 0 FAECIJ+URWPalladioL-BoldItal Type 1 Custom yes yes no 85 0 MDJLZU+EURM10 Type 1 Builtin yes yes no 92 0 NAHIBO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 106 0 NAHIDA+PalatinoLinotype TrueType WinAnsi yes yes no 109 0 NAHIEC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 112 0 VORBJF+CMSY10 Type 1 Builtin yes yes no 120 0 NAHIBO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 134 0 NAHICP+PalatinoLinotype CID TrueType Identity-H yes yes yes 137 0 NAHIDA+PalatinoLinotype TrueType WinAnsi yes yes no 143 0 NAHIEC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 146 0 NAHIBO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 161 0 NAHICP+PalatinoLinotype CID TrueType Identity-H yes yes yes 164 0 NAHIDA+PalatinoLinotype TrueType WinAnsi yes yes no 170 0 NAHIEC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 173 0 NAHIBO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 189 0 NAHICP+PalatinoLinotype CID TrueType Identity-H yes yes yes 192 0 NAHIDA+PalatinoLinotype TrueType WinAnsi yes yes no 198 0 NAHIEC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 201 0 NAHIBO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 217 0 NAHICP+PalatinoLinotype CID TrueType Identity-H yes yes yes 220 0 NAHIDA+PalatinoLinotype TrueType WinAnsi yes yes no 226 0 NAHIEC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 229 0 NAHIBO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 244 0 NAHICP+PalatinoLinotype CID TrueType Identity-H yes yes yes 247 0 NAHIDA+PalatinoLinotype TrueType WinAnsi yes yes no 253 0 NAHIEC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 256 0 NAIAJC+Arial,Bold CID TrueType Identity-H yes yes yes 259 0 NAHIBO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 275 0 NAHICP+PalatinoLinotype CID TrueType Identity-H yes yes yes 278 0 NAHIDA+PalatinoLinotype TrueType WinAnsi yes yes no 284 0 NAHIEC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 287 0 NAIAJC+Arial,Bold CID TrueType Identity-H yes yes yes 290 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2021-09-11 03:51:32 CEST RepresentationInformation: pmc_33498698.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2021-02-08 03:37:00 CET Size: 2302784 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 516 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Children: Item: Title: Patients Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: The Increase of B Cell Population during Sorafenib Treatment Increases the Probability of Developing eDAEs Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Lymphocytes Expressing DNAM-1 and PD-1 Are Significantly Associated with the Probability of Developing eDAEs, Both in Baseline and Time-Dependent Models Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: CD16, PD-1, and LAG-3 Are Co-Expressed in NK-Like CD3+ Cells and Correlate with eDAEs Development Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: T DNAM-1+ Cells Have an Exhausted Phenotype Compared to DNAM-1- Counterparts; While NK DNAM-1+ Cells Have an Immune Activated Phenotype Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: CD96, an NK Inhibitory Checkpoint, Is Highly Expressed in T and NK-Like CD3+ Cells, But Not in Conventional NK Cells Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: T CD4+ DNAM-1+ Cells Present Higher Immune-Suppressive Ability Ex Vivo Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Children: Item: Title: Patients Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Blood Extraction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: PBMCs Isolation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Flow Cytometry Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Antibodies and Reagents Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Cytokine Release Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Immune-Suppression Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Info: Title: Activated Lymphocytes and Increased Risk of Dermatologic Adverse Events during Sorafenib Therapy for Hepatocellular Carcinoma Author: Josep Corominas, Victor Sapena, Marco Sanduzzi-Zamparelli, Cristina Millán, Esther Samper, Neus Llarch, Gemma Iserte, Ferràn Torres, Leonardo G. Da Fonseca, Sergio Muñoz-Martínez, Alejandro Forner, Jordi Bruix, Loreto Boix and María Reig Subject: Advanced hepatocellular carcinoma patients treated with sorafenib who develop early dermatologic adverse events (eDAEs) have a better prognosis. This may be linked to immune mechanisms, and thus, it is relevant to assess the association between peripheral immunity and the probability of developing eDAEs. Peripheral blood mononuclear cells of 52 HCC patients treated with sorafenib were analyzed at baseline and throughout the first eight weeks of therapy. T, B, Natural Killer cells, and their immune checkpoints expression data were characterized by flow cytometry. Cytokine release and immune-suppression assays were carried out ex vivo. Cox baseline and time-dependent regression models were applied to evaluate the probability of increased risk of eDAEs. DNAM-1, PD-1, CD69, and LAG-3 in T cells, plus CD16 and LAG-3 in NK cells, are significantly associated with the probability of developing eDAEs. While NK DNAM-1+ cells express activation markers, T DNAM-1+ cells induce immune suppression and show immune exhaustion. This is the first study to report an association between immune checkpoints expression in circulating immune cells and the increased incidence of eDAEs. Our results support the hypothesis for an off-target role of sorafenib in immune modulation. We also describe a novel association between DNAM-1 and immune exhaustion in T cells. 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