Volume 54, Issue 2, Pages 570-579 (August 1998) Interleukin-6, interleukin-8 and monocyte chemotactic peptide-1 gene expression and protein synthesis are independently modulated by hemodialysis membranes  Giovanni Pertosa, Giuseppe Grandaliano, Loreto Gesualdo, Elena Ranieri, Raffaella Monno, F. Paolo Schena  Kidney International  Volume 54, Issue 2, Pages 570-579 (August 1998) DOI: 10.1046/j.1523-1755.1998.00020.x Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 1 Spontaneous production of IL-6 (A), IL-8 (B) and MCP-1 (C) from PBMC of normal subjects and uremic patients on conservative therapy. Cytokine release, particularly IL-8 and MCP-1, was reduced in the uremic group compared to the healthy subjects. *P < 0.05 and **P < 0.01 versus normal subjects. Kidney International 1998 54, 570-579DOI: (10.1046/j.1523-1755.1998.00020.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 2 (A) Spontaneous production of IL-6 from PBMC of normal subjects (N) and uremic patients before (T0; ▪) and after (T180; ) dialysis with different membranes. (B) IL-6 production from PBMC at T0 in any single patient included in the study; spontaneous production in normal subjects (N) is shown as mean ± sem. **P < 0.01 versus normal controls. Abbreviations are: CU, cuprophan; PMMA, polymethylmetacrylate; PA, polyamide. Kidney International 1998 54, 570-579DOI: (10.1046/j.1523-1755.1998.00020.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 3 (A) Spontaneous production of IL-8 from PBMC of normal subjects (N) and uremic patients before (T0; ▪) and after (T180; ) dialysis with different membranes. (B) IL-8 production from PBMC at T0 in any single patient included in the study; spontaneous production in normal subjects (N) is shown as mean ± sem. **P < 0.01 versus normal subjects. Abbreviations are: CU, cuprophan; PMMA, polymethylmetacrylate; PA, polyamide. Kidney International 1998 54, 570-579DOI: (10.1046/j.1523-1755.1998.00020.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 4 (A) Spontaneous production of MCP-1 from PBMC of normal subjects and uremic patients before (T0; ▪) and after (T180;) dialysis with different membranes. (B) MCP-1 production from PBMC at T0 in any single patient included in the study; spontaneous production in normal subjects (N) is shown as mean ± sem. **P < 0.01 versus normal subjects. Abbreviations are: CU, cuprophan; PMMA, polymethylmetacrylate; PA, polyamide. Kidney International 1998 54, 570-579DOI: (10.1046/j.1523-1755.1998.00020.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 5 (A) IL-6 (upper panel) and GAPDH (lower panel) gene expression by PBMC during hemodialysis with different membranes, studied by RT-PCR and Southern blotting as described in the Methods section (a representative gel is shown). (B) An increase of IL-6 mRNA expression was observed in uremic patients treated with CU as compared to normal subjects (N) and non-dialyzed uremic patients (U). Symbols are: (▪) T0, before dialysis; (□) T180, after dialysis. By contrast, the use of non-complement activating membranes, such as PMMA and PA, determined a reduced expression of IL-6 mRNA in pre-HD (T0) PBMC as compared to CU-treated patients.*P < 0.05 versus normal subjects; **P < 0.05 versus CU T0. Abbreviations are: CU, cuprophan; PMMA, polymethylmetacrylate; PA, polyamide. Kidney International 1998 54, 570-579DOI: (10.1046/j.1523-1755.1998.00020.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 6 (A) IL-8 (upper panel) and GAPDH (lower panel) gene expression by PBMC during hemodialysis with different membranes, studied by RT-PCR as described in the Methods section (a representative gel is shown). (B) An increase in IL-8 transcripts was observed in uremic patients treated with CU membranes, as compared to normal subjects (N) and non-dialyzed uremic patients (U). By contrast, the use of biocompatible membranes (PMMA and PA) determined a decrease in IL-8 mRNA levels.*P < 0.05 versus normal subjects; **P < 0.05 versus CU T0. Abbreviations are: CU, cuprophan; PMMA, polymethylmetacrylate; PA, polyamide. Kidney International 1998 54, 570-579DOI: (10.1046/j.1523-1755.1998.00020.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 7 (A) MCP-1 (upper panel) and GAPDH (lower panel) gene expression by PBMC during hemodialysis with different membranes, studied by RT-PCR as described in the Methods section (a representative gel is shown). (B) An increase in MCP-1 transcripts was observed in uremic patients treated with CU membranes, as compared to normal subjects (N) and non-dialyzed uremic patients (U). By contrast, the use of biocompatible membranes (PMMA and PA) determined a decrease in MCP-1 mRNA levels.*P < 0.05 versus normal subjects; **P < 0.05 versus CU T0. Abbreviations are: CU, cuprophan; PMMA, polymethylmetacrylate; PA, polyamide. Kidney International 1998 54, 570-579DOI: (10.1046/j.1523-1755.1998.00020.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 8 MCP-1 gene expression in PBMC of controls and hemodialysis patients treated with different membranes. MCP-1 gene expression was studied by in situ hybridization as described in the Methods section. Bright (A, C, E and G) and dark (B, D, F and H) field photomicrographs of PBMC isolated from healthy subjects (A and B) and at T0 from patients treated by CU (C and D), PMMA (E and F) and PA (G and H) membranes. CU treatment induces a clear upregulation of MCP-1 mRNA whereas the treatment with non-complement activating membranes reduces it. Kidney International 1998 54, 570-579DOI: (10.1046/j.1523-1755.1998.00020.x) Copyright © 1998 International Society of Nephrology Terms and Conditions