3feac52422f4f4424a867f8e3bf125b0 microorganisms-13-01164-v2_1.pdf 49bfd16dadea8fbd1c1cfb08567bbf0b7ab89cab microorganisms-13-01164-v2_1.pdf 484dccd592b64e6ee336db55a0a57e2ae72dd74e69ff5cb9d2fda9c78ba6bbed microorganisms-13-01164-v2_1.pdf Title: In Vitro Immune Response of Mononuclear Cells to Multidrug-Resistant Escherichia coli Subject: Infections caused by multidrug-resistant organisms (MDRO) are linked to poor outcomes, particularly in patients with cirrhosis. The underlying mechanisms are not fully understood and may involve a different immune response against MDRO. This study aimed to compare the in vitro immune response between multidrug-resistant (MDR) Escherichia coli and antibiotic-susceptible E. coli strains. Surface protein extract and DNA extract were obtained from MDR E. coli (n = 6) and antibiotic-susceptible E. coli (n = 6) strains isolated from infected patients with cirrhosis. The extracts were used to stimulate in vitro peripheral blood mononuclear cells from healthy donors. After 48 h, cytokine levels (IFN-, IL-1, IL-10, IL-12p70, MCP-1, IL-8, IL-6, MIP-1, and MIP-1) were measured. We observed no significant differences in cytokine production between MDR and susceptible strains. However, we identified notable interindividual variability in cytokine production for most of the cytokines studied. Only IFN- and IL-6 in surface extract and MCP-1 in DNA extract showed similar levels across all donors. We conclude that the cytokine profiles induced by MDR E. coli in vitro were similar to those in susceptible strains. These findings suggest that the poor prognosis associated with MDR E. coli infections is not due to a differential immune response but rather to other factors. Keywords: resistance; cirrhosis; immune response; MDRO; Escherichia coli; cytokines Author: Berta Cuyàs, Elisabet Cantó, Elisabet Sanchez-Ardid, Elisenda Miró, Edilmar Alvarado-Tapias, Eva Román, Maria Poca, Ferran Navarro, Andreu Ferrero-Gregori, Maria Àngels Escorsell, Silvia Vidal and German Soriano Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Wed May 21 12:19:18 2025 CEST ModDate: Wed May 21 12:22:14 2025 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 16 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 5140691 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- LEVXPH+EURM10 Type 1 Builtin yes yes yes 10 0 ODASUJ+VnURWPalladioL-Bold Type 1 Custom yes yes yes 15 0 XZVRGJ+URWPalladioL-Roma Type 1 Custom yes yes yes 21 0 NVWMOI+URWPalladioL-Bold Type 1 Custom yes yes yes 27 0 UQEDHB+URWPalladioL-Ital Type 1 Custom yes yes yes 32 0 RIOTWG+URWPalladioL-BoldItal Type 1 Custom yes yes yes 37 0 HQABBJ+VnURWPalladioL Type 1 Custom yes yes yes 42 0 QMDDXF+CMSY10 Type 1 Builtin yes yes yes 68 0 JHYTSG+CMR10 Type 1 Builtin yes yes yes 79 0 MFFDOG+PalatinoLinotype TrueType WinAnsi yes yes no 95 0 MFFDOE+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 300 0 MFFDOG+PalatinoLinotype TrueType WinAnsi yes yes no 303 0 MFFDOI+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 306 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-05-22 02:42:47 CEST RepresentationInformation: microorganisms-13-01164-v2_1.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-05-21 16:54:03 CEST Size: 5140691 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 576 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: Bacterial Isolates and Growth Conditions Destination: subsection.2.1 Item: Title: Bacterial Extracts Isolations Destination: subsection.2.2 Children: Item: Title: Surface Protein Isolation Destination: subsubsection.2.2.1 Item: Title: Bacterial DNA Isolation Destination: subsubsection.2.2.2 Item: Title: Peripheral Blood Mononuclear Cells (PBMC) Isolation and Stimulation with Bacterial Extracts Destination: subsection.2.3 Item: Title: Determination of Cytokine Concentration and Ratio Destination: subsection.2.4 Item: Title: Statistical Analyses Destination: subsection.2.5 Item: Title: Results Destination: section.3 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: appendix.A. Info: Title: In Vitro Immune Response of Mononuclear Cells to Multidrug-Resistant Escherichia coli Author: Berta Cuyàs, Elisabet Cantó, Elisabet Sanchez-Ardid, Elisenda Miró, Edilmar Alvarado-Tapias, Eva Román, Maria Poca, Ferran Navarro, Andreu Ferrero-Gregori, Maria Àngels Escorsell, Silvia Vidal and German Soriano Subject: Infections caused by multidrug-resistant organisms (MDRO) are linked to poor outcomes, particularly in patients with cirrhosis. The underlying mechanisms are not fully understood and may involve a different immune response against MDRO. This study aimed to compare the in vitro immune response between multidrug-resistant (MDR) Escherichia coli and antibiotic-susceptible E. coli strains. Surface protein extract and DNA extract were obtained from MDR E. coli (n = 6) and antibiotic-susceptible E. coli (n = 6) strains isolated from infected patients with cirrhosis. The extracts were used to stimulate in vitro peripheral blood mononuclear cells from healthy donors. After 48 h, cytokine levels (IFN-, IL-1, IL-10, IL-12p70, MCP-1, IL-8, IL-6, MIP-1, and MIP-1) were measured. We observed no significant differences in cytokine production between MDR and susceptible strains. However, we identified notable interindividual variability in cytokine production for most of the cytokines studied. Only IFN- and IL-6 in surface extract and MCP-1 in DNA extract showed similar levels across all donors. We conclude that the cytokine profiles induced by MDR E. coli in vitro were similar to those in susceptible strains. These findings suggest that the poor prognosis associated with MDR E. coli infections is not due to a differential immune response but rather to other factors. 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